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Background: Immunotherapy has emerged as an efficient therapeutic approach for cancer management. However, stimulation of host immune system against cancer cells often fails to achieve promising clinical outcomes mainly owing to the immunosuppressive characteristics of the tumor microenvironment (TME). Combination therapeutics that can trigger sustained immunogenic cell death (ICD) have provided new opportunities for cancer treatment. Methods: In this study, we designed and applied an ICD inducer regimen, including a genetically engineered oncolytic virus (miRNA-modified coxsackieviruses B3, miR-CVB3), a pore-forming lytic peptide (melittin, found in bee venom), and a synthetic toll-like receptor 9 ligand (CpG oligodeoxynucleotides), for breast cancer and melanoma treatment. We compared the anti-tumor efficacy of miR-CVB3 and CpG-melittin (CpGMel) alone and in combination (miR-CVB3 + CpGMel) and investigated possible mechanisms involved. Results: We demonstrated that miR-CVB3 + CpGMel had no major impact on viral growth, while enhancing the cellular uptake of CpGMel in vitro. We further showed that combination therapy led to significant increases in tumor cell death and release of damage-associated molecular patterns compared with individual treatment. In vivo studies in 4T1 tumor-bearing Balb/c mice revealed that both primary and distant tumors were significantly suppressed, and the survival rate was significantly prolonged after administration of miR-CVB3 + CpGMel compared with single treatment. This anti-tumor effect was accompanied by increased ICD and immune cell infiltration into the TME. Safety analysis showed no significant pathological abnormalities in Balb/c mice. Furthermore, the developed therapeutic regimen also demonstrated a great anti-tumor activity in B16F10 melanoma tumor-bearing C57BL/6 J mice. Conclusions: Overall, our findings indicate that although single treatment using miR-CVB3 or CpGMel can efficiently delay tumor growth, combining oncolytic virus-based therapy can generate even stronger anti-tumor immunity, leading to a greater reduction in tumor size. [ABSTRACT FROM AUTHOR]

Ehrlichia chaffeensis, a tick-transmitted intraphagosomal bacterium, is the causative agent of human monocytic ehrlichiosis. The pathogen also infects several other vertebrate hosts. E. chaffeensis has a biphasic developmental cycle during its growth in vertebrate monocytes/macrophages and invertebrate tick cells. Host- and vector-specific differences in the gene expression from many genes of E. chaffeensis are well documented. It is unclear how the organism regulates gene expression during its developmental cycle and for its adaptation to vertebrate and tick host cell environments. We previously mapped promoters of several E. chaffeensis genes which are recognized by its only two sigma factors: σ32 and σ70. In the current study, we investigated in assessing five predicted E. chaffeensis transcription regulators; EcxR, CtrA, MerR, HU and Tr1 for their possible roles in regulating the pathogen gene expression. Promoter segments of three genes each transcribed with the RNA polymerase containing σ70 (HU, P28-Omp14 and P28-Omp19) and σ32 (ClpB, DnaK and GroES/L) were evaluated by employing multiple independent molecular methods. We report that EcxR binds to all six promoters tested. Promoter-specific binding of EcxR to several gene promoters results in varying levels of gene expression enhancement. This is the first detailed molecular characterization of transcription regulators where we identified EcxR as a gene regulator having multiple promoter-specific interactions. [ABSTRACT FROM AUTHOR]

Background: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron system associated with both genetic and environmental risk factors. Infection with enteroviruses, including poliovirus and coxsackievirus, such as coxsackievirus B3 (CVB3), has been proposed as a possible causal/risk factor for ALS due to the evidence that enteroviruses can target motor neurons and establish a persistent infection in the central nervous system (CNS), and recent findings that enteroviral infection-induced molecular and pathological phenotypes closely resemble ALS. However, a causal relationship has not yet been affirmed.Methods: Wild-type C57BL/6J and G85R mutant superoxide dismutase 1 (SOD1G85R) ALS mice were intracerebroventricularly infected with a sublethal dose of CVB3 or sham-infected. For a subset of mice, ribavirin (a broad-spectrum anti-RNA viral drug) was given subcutaneously during the acute or chronic stage of infection. Following viral infection, general activity and survival were monitored daily for up to week 60. Starting at week 20 post-infection (PI), motor functions were measured weekly. Mouse brains and/or spinal cords were harvested at day 10, week 20 and week 60 PI for histopathological evaluation of neurotoxicity, immunohistochemical staining of viral protein, neuroinflammatory/immune and ALS pathology markers, and NanoString and RT-qPCR analysis of inflammatory gene expression.Results: We found that sublethal infection (mimicking chronic infection) of SOD1G85R ALS mice with CVB3 resulted in early onset and progressive motor dysfunction, and shortened lifespan, while similar viral infection in C57BL/6J, the background strain of SOD1G85R mice, did not significantly affect motor function and mortality as compared to mock infection within the timeframe of the current study (60 weeks PI). Furthermore, we showed that CVB3 infection led to a significant increase in proinflammatory gene expression and immune cell infiltration and induced ALS-related pathologies (i.e., TAR DNA-binding protein 43 (TDP-43) pathology and neuronal damage) in the CNS of both SOD1G85R and C57BL/6J mice. Finally, we discovered that early (day 1) but not late (day 15) administration of ribavirin could rescue ALS-like neuropathology and symptoms induced by CVB3 infection.Conclusions: Our study identifies a new risk factor that contributes to early onset and accelerated progression of ALS and offers opportunities for the development of novel targeted therapies. [ABSTRACT FROM AUTHOR]

We synthesized ppy@DR-MoS 2 as sulfur host to improve the performance of lithium-sulfur batteries by adopting a collaborative strategy of structural design and defect engineering. The flexible ppy can suppress the volume changes of sulfur during charging and discharging and adsorb LiPSs physically. Furthermore, the abundant S defects make MoS 2 with enhanced chemisorption of polysulfides, and excellent electrocatalytic activity. [Display omitted] • Nanoflowers structure was built; • Sulfur defect can improve the kinetics of lithium-sulfur batteries; • PPy coating blocks the sulfurous shuttle. The shuttle effect and the slow conversion kinetics hinder the practical application of lithium-sulfur batteries. Herein, we synthesized the polypyrrole (PPy) coated defect-rich MoS 2 nanoflowers (PPy@DR-MoS 2) as sulfur host. Benefiting from the sulfur defects and PPy coating structure, the S/ppy-DR-MoS 2 composite displays good cycling performance at a high current density of 1C and excellent cycling stability for over 300 cycles with a low cycling decline rate of 0.17% per cycle. [ABSTRACT FROM AUTHOR]

The alfalfa harvest time is determined by the alfalfa growth stage and the weather factors at that time, which will affect the yield and quality of the alfalfa hay. Addressing issues like the lack of precise harvesting timing and underutilized production potential of lucerne, this study is based on the internal(fall dormancy level, crude protein, acid, neutral detergent fiber, relative feed value) and external factors(air temperature, precipitation, wind speed, ground temperature, relative humidity, solar radiation) affecting the quality of lucerne, as well as the relevant experimental data of the existing literature and the weather factors within 24 h after the lucerne harvest.Using principal component analysis to calculate the weights of the above indicators, the harvest suitability score formula was constructed to determine the harvesting time, harvesting method and harvesting stubble. Huanghua City was used as an example to confirm the method's effectiveness in guiding for determining the optimal harvest date of lucerne in the city. Key findings include the following: The hay quality of the first two crops of alfalfa in Huanghua City was higher than that of the third and fourth crops, and the optimal harvest stage of alfalfa was before the precipitation concentration period, which was consistent with other local research results and weather characteristics, and the method was feasible. Precipitation was the most significant factor affecting the lucerne harvest. The effects of other weather factors on hay quality varied with precipitation conditions. The temperature was the second important factor when daily precipitation is less than 1 mm, and the wind speed became the second important factor when the daily precipitation is less than 10 mm and more than 1 mm. Under low suitability and high precipitation, it may be advisable to convert hay harvest to silage to maximize economic benefits. This research could help agricultural decision-makers to develop precise harvesting strategies, and further improve lucerne production efficiency. [ABSTRACT FROM AUTHOR]

Electrochemiluminescence (ECL) sensors are useful for the detection of heavy metal pollutants, in particular mercury(ii) ions, in water samples. We demonstrate the superior sensing performance of Hg2+ using a nanocomposite material based on carbon nitride nanosheets (CNNSs) and copper nanoclusters functionalized by dithiothreitol, which not only stabilizes the clusters, but also improves the sensitivity of Hg2+ detection. The ECL mechanism is related to the reaction of the nanocomposite with K2S2O8 in the electrochemical system, while the presence of Hg2+ leads to quenching of its excited state, and the suppression of the formation of anion-radicals. The Hg(ii) sensor presented here is cheap and fast, and shows high selectivity for the detection of Hg2+ on the background of other mono-, di-, and trivalent ions, with a linear range of 0.5–10 nM and the detection limit as low as 0.01 nM. [ABSTRACT FROM AUTHOR]

Coxsackievirus B3 (CVB3) displays great oncolytic activity against various cancer cells. Previously, we demonstrated that adding targeting sequences (TS) of miR-145/143, which are downregulated in cancer compared with normal cells, into CVB3 genome drastically attenuates tissue toxicity, while retaining its oncolytic activity towards lung tumor. Here we extended to assess miR-modified CVB3 in breast cancer therapy. We generated a new miRNA-CVB3 by inserting TS of muscle-specific miR-1 and pancreas-selective miR-216 into the above miR-145/143-modified CVB3. We found that this newly established CVB3 (termed miR-CVB3-1.1) is safe without triggering noticeable pathogenesis when applied to immunocompetent mice. In vitro studies revealed that miR-CVB3-1.1 can infect and lyse a wide range of breast cancer cells. Animal experiments using a syngeneic breast cancer mouse model showed that intratumoral inoculation of miR-CVB3-1.1 significantly suppresses tumor growth and metastasis, associated with productive viral growth and enhanced immune cell infiltration in the tumor microenvironment. Moreover, we observed substantially reduced toxicity and prolonged survival in mice treated with miR-CVB3-1.1 compared with wild-type CVB3. Together, our results support miR-CVB3-1.1 as a promising candidate, which can be further evaluated for clinical treatment of breast cancer. [ABSTRACT FROM AUTHOR]

We synthesized thiolated few-layered graphene (FLG) on a Cu substrate by thermal chemical vapor deposition using thiocamphor as the solid precursor of both the carbon and sulfur. The Raman mapping results and transmission electron microscopy (TEM) images identified the numbers of layers and the morphology of the thiolated graphene sheets. The growth of few-layered including mono- and bi-layered thiolated graphene was verified by Raman mapping. TEM observation confirmed formation of bi- and nine-layers of thiolated graphene. X-ray photoelectron spectroscopy (XPS) revealed that the synthesized graphene was terminated with thiol group. The concentration of sulfur atoms in the thiolated FLG sample synthesized from thiocamphor was evaluated to be 7.69% by XPS. The environmental stability of the thiolated FLG was also investigated by performing XPS on the samples after several days. The S2p peak was not detectable after 30 days. [ABSTRACT FROM AUTHOR]

In this paper, polyacrylamide modified graphene oxide (PAM/GO) composites were synthesized and characterized by scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, Raman spectroscopy, and X-ray diffraction. The surface site distribution of PAM/GO was simulated with the aid of FITEQL 3.2 code. The synthesized PAM/GO composites were applied to adsorb Sr(II) from aqueous solutions. The adsorption conditions, such as contact time, pH value, and ionic strength, were investigated, which indicated pH and ionic strength dependent adsorption process. The adsorption isotherm agreed well with the Langmuir model, with a maximum adsorption capacity of 2.11 mmol/g at pH = 8.5 ± 0.1 and T = 303 K. Thermodynamic parameters calculated from the temperature-dependent adsorption isotherms suggested that the adsorption of Sr(II) on PAM/GO composites was an endothermic and spontaneous process. The efficient adsorption suggested that PAM/GO might be a potential candidate for the preconcentration and separation of Sr(II) from wastewater. [ABSTRACT FROM AUTHOR]

Highlights: [•] Oat has the potential to meliorate the saline-alkali soil by high ion concentration. [•] Oat phytoremediation capability affected by soil salinity, harvest time and varieties. [•] Delayed harvest significantly declines the straw ion content of Na+, K+, Cl−. [•] Soil salinity enhances straw ion concentration but reduced its accumulation. [•] Stem total ion content is several times higher than the ion content in soil. [Copyright &y& Elsevier]

TiO/MCM-41 composites with various titania content were prepared by loading titania into the mesopores of MCM-41 molecular sieves by sol-gel method, and were used as photocatalysts to degrade Rhodamine B (RhB) and phenol. The efficiency of organic contaminants removal was increased significantly compared with pure TiO. Ti/Si ratio, namely, the content of TiO was determined by ICP-AES method. TiO/MCM-41 composites were characterized by X-ray diffraction, UV-Vis absorption spectroscopy and N adsorption techniques. Experimental results demonstrated that most of the RhB was adsorbed instead of being degradated by TiO/MCM-41 due to the large specific surface area of MCM-41, while most of phenol was degradated. It turned out that the TiO/MCM-41 with the highest Ti/Si ratio of 0.8220 (wt) had the highest catalytic activity. [ABSTRACT FROM AUTHOR]

Background: Plant mitochondria, semiautonomous organelles that function as manufacturers of cellular ATP, have their own genome that has a slow rate of evolution and rapid rearrangement. Cytoplasmic male sterility (CMS), a common phenotype in higher plants, is closely associated with rearrangements in mitochondrial DNA (mtDNA), and is widely used to produce F1 hybrid seeds in a variety of valuable crop species. Novel chimeric genes deduced from mtDNA rearrangements causing CMS have been identified in several plants, such as rice, sunflower, pepper, and rapeseed, but there are very few reports about mtDNA rearrangements in wheat. In the present work, we describe the mitochondrial genome of a wheat K-type CMS line and compare it with its maintainer line. Results: The complete mtDNA sequence of a wheat K-type (with cytoplasm of Aegilops kotschyi) CMS line, Ks3, was assembled into a master circle (MC) molecule of 647,559 bp and found to harbor 34 known protein-coding genes, three rRNAs (18 S, 26 S, and 5 S rRNAs), and 16 different tRNAs. Compared to our previously published sequence of a K-type maintainer line, Km3, we detected Ks3-specific mtDNA (> 100 bp, 11.38%) and repeats (> 100 bp, 29 units) as well as genes that are unique to each line: rpl5 was missing in Ks3 and trnH was absent from Km3. We also defined 32 single nucleotide polymorphisms (SNPs) in 13 protein-coding, albeit functionally irrelevant, genes, and predicted 22 unique ORFs in Ks3, representing potential candidates for K-type CMS. All these sequence variations are candidates for involvement in CMS. A comparative analysis of the mtDNA of several angiosperms, including those from Ks3, Km3, rice, maize, Arabidopsis thaliana, and rapeseed, showed that non-coding sequences of higher plants had mostly divergent multiple reorganizations during the mtDNA evolution of higher plants. Conclusion: The complete mitochondrial genome of the wheat K-type CMS line Ks3 is very different from that of its maintainer line Km3, especially in non-coding sequences. Sequence rearrangement has produced novel chimeric ORFs, which may be candidate genes for CMS. Comparative analysis of several angiosperm mtDNAs indicated that non-coding sequences are the most frequently reorganized during mtDNA evolution in higher plants. [ABSTRACT FROM AUTHOR]

Abstract: Quantitative structure–activity relationship (QSAR) model was used to predict and explain binding constant (log K) determined by fluorescence quenching. This method allowed us to predict binding constants of a variety of compounds with human serum albumin (HSA) based on their structures alone. Stepwise multiple linear regression (MLR) and nonlinear radial basis function neural network (RBFNN) were performed to build the models. The statistical parameters provided by the MLR model (R 2=0.8521, RMS=0.2678) indicated satisfactory stability and predictive ability while the RBFNN predictive ability is somewhat superior (R 2=0.9245, RMS=0.1736). The proposed models were used to predict the binding constants of two bioactive components in traditional Chinese medicines (isoimperatorin and chrysophanol) whose experimental results were obtained in our laboratory and the predicted results were in good agreement with the experimental results. This QSAR approach can contribute to a better understanding of structural factors of the compounds responsible for drug–protein interactions, and can be useful in predicting the binding constants of other compounds. [ABSTRACT FROM AUTHOR]

Background: Ehrlichia chaffeensis is a tick-transmitted rickettsial pathogen responsible for an important emerging disease, human monocytic ehrlichiosis. To date how E. chaffeensis and many related tick-borne rickettsial pathogens adapt and persist in vertebrate and tick hosts remain largely unknown. In recent studies, we demonstrated significant host-specific differences in protein expression in E. chaffeensis originating from its tick and vertebrate host cells. The adaptive response of the pathogen to different host environments entails switch of gene expression regulated at the level of transcription, possibly by altering RNA polymerase activity. Results: In an effort to understand the molecular basis of pathogen gene expression differences, we isolated native E. chaffeensis RNA polymerase using a heparin-agarose purification method and developed an in vitro transcription system to map promoter regions of two differentially expressed genes of the p28 outer membrane protein locus, p28-Omp14 and p28-Omp19. We also prepared a recombinant protein of E. chaffeensis σ70 homologue and used it for in vitro promoter analysis studies. The possible role of one or more proteins presents in E. chaffeensis lysates in binding to the promoter segments and on the modulation of in vitro transcription was also assessed. Conclusions: Our experiments demonstrated that both the native and recombinant proteins are functional and have similar enzyme properties in driving the transcription from E. chaffeensis promoters. This is the first report of the functional characterization of E. chaffeensis RNA polymerase and in vitro mapping of the pathogen promoters using the enzyme. This study marks the beginning to broadly characterize the mechanisms controlling the transcription by Anaplasmataceae pathogens. [ABSTRACT FROM AUTHOR]

Abstract: Quantitative structure–property relationship models correlating the half-wave potentials (E 1/2) of the dye intermediate 9, 10-anthraquinone and its derivatives were developed using both linear and non-linear modelling approaches. Descriptors calculated from molecular structures alone were used to represent the E 1/2 of the 9, 10-anthraquinones. An heuristic method was used to select the most appropriate molecular descriptors whilst a linear, quantitative structure–property relationship model was developed; using the selected descriptors, a radial basis function neural network was employed for the non-linear model development. The statistical parameters provided by the heuristic model were R 2 = 0.945; F = 98.04; RMS = 0.0360 for the training set and R 2 = 0.863; F = 18.903; RMS = 0.0600 for the test set. The radial basis function neural network model gave better results with R 2 = 0.960; F = 610.336; RMS = 0.0299 for the training set and R 2 = 0.863; F = 18.946; RMS = 0.0476 for the test set. [Copyright &y& Elsevier]

Abstract: QSPR models for the prediction of UV maximum absorption wavelength (λ max) of 69 flavones were developed based on their structures alone. A six-descriptor linear correlation by heuristic method (HM) and a nonlinear model using radial basis function neural network (RBFNN) approach were reported. The statistical parameters provided by the HM model (R 2 =0.961, F =207.820, RMS=6.555 for the training set and R 2 =0.967, F =293.218, RMS=7.176 for the test set) and the RBFNN model (R 2 =0.971, F =1826.086, RMS=5.350 for the training set, and R 2 =0.978, F =452.512, RMS=5.722 for the test set) indicated satisfactory stability and predictive ability. The descriptors appearing in these models are discussed. This QSPR approach is suitable for the prediction of maximum absorption wavelength of flavones, and can contribute to a better understanding of structural factors of the organic compounds responsible for it. [Copyright &y& Elsevier]

Abstract: A quantitative structure–property relationship study was performed on the hydrophile–lipophile balance (HLB) values of anionic surfactants. Stepwise multiple linear regression (MLR) and nonlinear radial basis function neural network (RBFNN) were used to build the models. A four-descriptor equation with the squared correlation coefficient of 0.983 and root mean square error (RMS) of 1.7309 were obtained for the training set, and , for the external test set. The RBFNN model gave better results: , for the training set and , for test set. The QSPR model established may provide a new powerful method for predicting HLB values of anionic surfactants. [Copyright &y& Elsevier]

Abstract: Orthogonal design has been used to the optimization of separation and determination of two active components in traditional Chinese medicines by capillary electrophoresis. The concentration of phosphate, applied voltage, organic modifier content and buffer pH were selected as variable parameters. Their different effects on peak resolution were studied by the experimental design method. Optimized separation conditions were obtained and successfully applied to the separation and determination of aconitine and hypaconitine in Aconitum medicinal herbs. Good separation was achieved within 7min using a buffer system composed of 20mmolL−1 phosphate and 35% acetonitrile at pH 9.5. The applied voltage was 14kV and the detection was set at 235nm. In addition, a radial basis function neural network with a “4-18-1” structure was developed based on the experimental results of orthogonal design and uniform design, and was applied to the prediction of peak resolution of the two active components under the optimum separation conditions given by orthogonal design. The predicted results were in good agreement with the experimental values, indicating that radial basis function neural network is a potential way for the selection of separation conditions in capillary electrophoresis. [Copyright &y& Elsevier]

Ehrlichia chaffeensis causes human monocytic ehrlichiosis. Little is known about how this and other related tick-borne rickettsia pathogens maintain pH homeostasis in acidified phagosomes and the extracellular milieu. The membrane-bound sodium (cation)/proton antiporters are found in a wide range of organisms aiding pH homeostasis. We recently reported a mutation in an antiporter gene of E. chaffeensis (ECH_0379) which causes bacterial in vivo attenuation. The E. chaffeensis genome contains 10 protein coding sequences encoding for predicted antiporters. We report here that nine of these genes are transcribed during the bacterial growth in macrophages and tick cells. All E. chaffeensis antiporter genes functionally complemented antiporter deficient Escherichia coli. Antiporter activity for all predicted E. chaffeensis genes was observed at pH 5.5, while gene products of ECH_0179 and ECH_0379 were also active at pH 8.0, and ECH_0179 protein was complemented at pH 7.0. The antiporter activity was independently verified for the ECH_0379 protein by proteoliposome diffusion analysis. This is the first description of antiporters in E. chaffeensis and demonstrates that the pathogen contains multiple antiporters with varying biological functions, which are likely important for the pH homeostasis of the pathogen's replicating and infectious forms. [ABSTRACT FROM AUTHOR]

Abstract: A new suspended photocatalyst, TiO2 and activated carbon (AC) particles immobilized on silicone rubber (SR) film, was prepared using a novel method and its photocatalytic activity on the photodegradation of acridine dye and phenol in aqueous solution was tested. Silicone rubber film used as the support has many advantages: it is transparent to light, easily to be recycled and may be suspended in the aqueous solution to increase the collision possibility of pollutants with TiO2 photocatalysts. The activated carbon used as adsorbent helps to accelerate the degradation of organic substrates. This immobilized hybrid catalyst 90TAS (90wt% TiO2–10wt% AC/SR) is efficient in photoinduced mineralization of organic substrates at dilute concentrations and can be recycled easily. [Copyright &y& Elsevier]

Oncolytic viruses have emerged as a promising strategy for cancer therapy due to their dual ability to selectively infect and lyse tumor cells and to induce systemic anti-tumor immunity. Among various candidate viruses, coxsackievirus group B (CVBs) have attracted increasing attention in recent years. CVBs are a group of small, non-enveloped, single-stranded, positive-sense RNA viruses, belonging to species human Enterovirus B in the genus Enterovirus of the family Picornaviridae. Preclinical studies have demonstrated potent anti-tumor activities for CVBs, particularly type 3, against multiple cancer types, including lung, breast, and colorectal cancer. Various approaches have been proposed or applied to enhance the safety and specificity of CVBs towards tumor cells and to further increase their anti-tumor efficacy. This review summarizes current knowledge and strategies for developing CVBs as oncolytic viruses for cancer virotherapy. The challenges arising from these studies and future prospects are also discussed in this review. [ABSTRACT FROM AUTHOR]

The ongoing pandemic of COVID-19 alongside the outbreaks of SARS in 2003 and MERS in 2012 underscore the significance to understand betacoronaviruses as a global health challenge. SARS-CoV-2, the etiological agent for COVID-19, has infected over 50 million individuals' worldwide with more than ∼1 million fatalities. Autophagy modulators have emerged as potential therapeutic candidates against SARS-CoV-2 but recent clinical setbacks urge for better understanding of viral subversion of autophagy. Using MHV-A59 as a model betacoronavirus , time-course infections revealed significant loss in the protein level of ULK1, a canonical autophagy-regulating kinase, and the concomitant appearance of a possible cleavage fragment. To investigate whether virus-encoded proteases target ULK1, we conducted in-vitro and cellular cleavage assays and identified ULK1 as a novel bona fide substrate of SARS-CoV-2 papain-like protease (PLpro). Mutagenesis studies discovered that ULK1 is cleaved at a conserved PLpro recognition sequence (LGGG) after G499, separating its N-terminal kinase domain from a C-terminal substrate recognition region. Over-expression of SARS-CoV-2 PLpro is sufficient to impair starvation-induced autophagy and disrupt formation of ULK1-ATG13 complex. Finally, we demonstrated a dual role for ULK1 in MHV-A59 replication, serving a pro-viral functions during early replication that is inactivated at late stages of infection. In conclusion, our study identified a new mechanism by which PLpro of betacoronaviruses induces viral pathogenesis by targeting cellular autophagy. • ULK1 is a novel target of betacoronavirus. • SARS-COV-2 papain-like protease (PLpro) cleaves ULK1 after G499. • PLpro disrupts ULK1 complex and cellular autophagy. • ULK1 is required early but dispensable during late MHV-A59 infection. [ABSTRACT FROM AUTHOR]

BACKGROUND: Natamycin is often added to pastries, cheeses, and beverages. The residual amount of natamycin should be less than 10 mg kg−1. The current method for its determination in various foodstuffs is high‐performance liquid chromatography (HPLC). Capillary electrophoresis (CE) is a simple, fast, and environmentally friendly method with low reagent consumption and comparable separation performance. However, no reports were found on the determination of natamycin in foods by CE. A CE method to determine natamycin is therefore sought. RESULTS: Natamycin in foods was determined by the capillary zone electrophoresis (CZE) method with ultraviolet‐visible (UV) detection. Separation conditions were optimized as 20 mM Na2HPO4, pH 9.2, with 25 kV applied voltage, and UV detection at 306 nm. Under optimal conditions, electrophoretic analysis was completed in less than 4 min, with a limit of detection (LOD) of 0.065 μg mL−1 and limit of quantitation (LOQ) of 0.22 μg mL−1. A good linear relationship (r2 = 0.999) was obtained at the range of 0.1–25 μg mL−1. A comparison with the HPLC‐UV method was also carried out according to the National Standards of the People's Republic of China. CONCLUSION: The results obtained by the CZE and HPLC methods are comparable but the proposed CZE method can help us obtain a shorter detection time at low cost. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Aim: Multiple sclerosis (MS) is an autoimmune disease, and its typical characteristics are neuroinflammation and the demyelination of neurons in the central nervous system (CNS). Sterile alpha and TIR motif containing 1 (SARM1) is an essential factor mediating axonal degeneration and SARM1 deletion reduces the neuroinflammation in spinal cord injury. This study aimed to explore the roles of SARM1 and its underlying mechanisms in MS. Methods: Experimental autoimmune encephalomyelitis (EAE, a model of MS) model was established. Immunostaining, western blot, electron microscope, and HE staining were used to examine the pathological manifestations such as inflammation, demyelination, and neuronal death in SARM1f/f EAE mice and SARM1Nestin‐CKO EAE mice. In addition, RNA‐seq, real‐time PCR and double‐immunostaining were used to examine the underlying mechanism of SARM1 in EAE mice. Results: SARM1 was upregulated in neurons of the spinal cords of EAE mice. SARM1 knockout in CNS ameliorated EAE with less neuroinflammation, demyelination, and dead neurons. Mechanically, SARM1 knockout resulted in the reduction of insulin‐like growth factor (IGF)‐binding protein 2 (IGFBP2) in neurons of EAE mice, which might inhibit the neuroinflammation through inhibiting NF‐κB signaling. Finally, activation of NF‐κB partially aggravated the neuroinflammation and demyelination deficits of SARM1Nestin‐CKO EAE mice. Conclusions: These results identified the unknown role of SARM1 in the promotion of neuroinflammation and demyelination and revealed a novel drug target pathway of SARM1/IGFBP2/NF‐κB for MS. [ABSTRACT FROM AUTHOR]

Bacterial gene transcription is initiated by RNA polymerase containing a sigma factor. To understand gene regulation in Ehrlichia chaffeensis, an important tick-transmitted rickettsiae responsible for human monocytic ehrlichiosis, we initiated studies evaluating the transcriptional machinery of several genes of this organism. We mapped the transcription start sites of 10 genes and evaluated promoters of five genes (groE, dnaK, hup, p28-Omp14 and p28-Omp19 genes). We report here that the RNA polymerase binding elements of E. chaffeensis gene promoters are highly homologous for its only two transcription regulators, sigma 32 and sigma 70, and that gene expression is accomplished by either of the transcription regulators. RNA analysis revealed that although transcripts for both sigma 32 and sigma 70 are upregulated during the early replicative stage, their expression patterns remained similar for the entire replication cycle. We further present evidence demonstrating that the organism’s -35 motifs are essential to transcription initiations. The data suggest that E. chaffeensis gene regulation has evolved to support the organism’s growth, possibly to facilitate its intraphagosomal growth. Considering the limited availability of genetic tools, this study offers a novel alternative in defining gene regulation in E. chaffeensis and other related intracellular pathogens. [ABSTRACT FROM AUTHOR]

The abuse and residues of antibiotics have a great impact on the environment and organisms, and their determination has become very important. Due to their low contents, varieties and complex matrices, effective recognition, separation and enrichment are usually required prior to determination. Molecularly imprinted polymers (MIPs), a kind of highly selective polymer prepared via molecular imprinting technology (MIT), are used widely in the analytical detection of antibiotics, as adsorbents of solid-phase extraction (SPE) and as recognition elements of sensors. Herein, recent advances in MIPs for antibiotic residue analysis are reviewed. Firstly, several new preparation techniques of MIPs for detecting antibiotics are briefly introduced, including surface imprinting, nanoimprinting, living/controlled radical polymerization, and multi-template imprinting, multi-functional monomer imprinting and dummy template imprinting. Secondly, several SPE modes based on MIPs are summarized, namely packed SPE, magnetic SPE, dispersive SPE, matrix solid-phase dispersive extraction, solid-phase microextraction, stir-bar sorptive extraction and pipette-tip SPE. Thirdly, the basic principles of MIP-based sensors and three sensing modes, including electrochemical sensing, optical sensing and mass sensing, are also outlined. Fourthly, the research progress on molecularly imprinted SPEs (MISPEs) and MIP-based electrochemical/optical/mass sensors for the detection of various antibiotic residues in environmental and food samples since 2018 are comprehensively reviewed, including sulfonamides, quinolones, β-lactams and so on. Finally, the preparation and application prospects of MIPs for detecting antibiotics are outlined. [ABSTRACT FROM AUTHOR]

Traditional Chinese medicine (TCM) is one of the most internationally competitive industries. In the context of TCM modernization and internationalization, TCM-related research studies have entered a fast track of development. At the same time, research of TCM is also faced with challenges, such as matrix complexity, component diversity and low level of active components. As an interdisciplinary technology, molecular imprinting technology (MIT) has gained popularity in TCM study, owing to the produced molecularly imprinted polymers (MIPs) possessing the unique features of structure predictability, recognition specificity and application universality, as well as physical robustness, thermal stability, low cost and easy preparation. Herein, we comprehensively review the recent advances of MIT for TCM studies since 2017, focusing on two main aspects including extraction/separation and purification and detection of active components, and identification analysis of hazardous components. The fundamentals of MIT are briefly outlined and emerging preparation techniques for MIPs applied in TCM are highlighted, such as surface imprinting, nanoimprinting and multitemplate and multifunctional monomer imprinting. Then, applications of MIPs in common active components research including flavonoids, alkaloids, terpenoids, glycosides and polyphenols, etc. are respectively summarized, followed by screening and enantioseparation. Related identification detection of hazardous components from TCM itself, illegal addition, or pollution residues (e.g., heavy metals, pesticides) are discussed. Moreover, the applications of MIT in new formulation of TCM, chiral drug resolution and detection of growing environment are summarized. Finally, we propose some issues still to be solved and future research directions to be expected of MIT for TCM studies. [ABSTRACT FROM AUTHOR]

An organic–inorganic polypyrrole/titanium(IV) biphosphate hybrid, PPy/Ti(HPO 4 ) 2 , was successfully synthesized by oxidative polymerization and applied to remove Cr(VI) from aqueous solutions. The as-prepared organic–inorganic hybrid was characterized by scanning electron microscope, X-ray diffraction, zeta potential analysis and X-ray photoelectron spectroscopy. The adsorption conditions, such as contact time, pH value and ionic strength, were investigated, indicating a pH and ionic strength dependent adsorption process. The adsorption kinetics fitted well with the pseudo-second-order kinetic model and the equilibrium data can be described by the Langmuir isotherm with a maximum adsorption capacity of 1.67 mmol/g at pH = 3.5 ± 0.1 and T = 288.15 K. Thermodynamic parameters calculated from the temperature-dependent adsorption isotherms suggested that the adsorption of Cr(VI) on PPy/Ti(HPO 4 ) 2 hybrid was an endothermic and spontaneous process. These results suggested that the PPy/Ti(HPO 4 ) 2 hybrid might be a potential candidate for the separation of Cr(VI) from aqueous solution. [ABSTRACT FROM AUTHOR]

[Display omitted] In this study, we constructed a metal-free Electro-Fenton system mediated by nitrogen-doped activated carbon modified graphite felt (NACs/GF) for degrading sulfamethazine (SMZ) through self-generation and utilization of H 2 O 2. With the increase of activation temperature (700 ∼ 1100 ℃), graphitic N occupied the dominant position and whose nearest pyridinic N were released to form N vacancies. NAC-1100/GF E-F system had a high H 2 O 2 selectivity (94.3 %) and an excellent H 2 O 2 yield (44.6 mg·L−1). The OOH* as the main barrier of 2e−-ORR process was formed and desorbed at graphitic N sites, while the N vacancies could enhance electron transfer with O 2 * and OOH* for accelerating generation of H 2 O 2. The 50 mg·L−1 of SMZ as pollutant could be completely degraded within 60 min in NAC-1100/GF E-F system, during which the graphitic N and pyrrolic N adsorbed SMZ through van der Waals forces, and pyridinic N adsorbed H 2 O 2 by hydrogen bonding interactions, thus establishing a controlled reaction zone near the cathode. The highly electronegativity pyridinic N could provide electrons for catalyzing the generation of ·OH from H 2 O 2 , while N vacancies could form more unpaired electrons to facilitate the formation of 1O 2. Finally, the degradation pathway of SMZ and toxicity analysis of degradation products were further clarified. This study provided valuable information for constructing metal-free Electro-Fenton systems and their application in environmental purification. [ABSTRACT FROM AUTHOR]

Preparation of high internal phase Pickering emulsions (HIPPEs) from starch nanocrystals (SNCs) is currently a key area of research. However, the high cost of producing SNCs restricts their usage in the food industry. This study investigated the effectiveness of micron-sized esterified maize starch (M-NMS) as the sole stabilizer for the preparation of HIPPEs. It was found that stable, gel-like HIPPEs with an oil phase volume of 75–90% could be created by using M-NMS granules at a concentration of ≥3.0 wt%. At a pH of 5.0–9.0, the absolute value of zeta potential for all emulsions exceeded 61 mV, indicating high interfacial stability. The addition of NaCl at a concentration of 40–160 mmol/l had no significant effect on the emulsion's viscosity, viscoelasticity, or stability. The gel structure formed by the swollen M-NMS molecular chain predominantly determined the emulsion's performance, as shown by atomic force microscopy. • Pickering emulsions of 75–90 vol% were prepared by using micron-scale starch. • A physical barrier was obtained by starch swelling at the oil drop interface. • In the pH values of 5.0–9.0, zeta potential of all emulsions exceeded 61 mV. [ABSTRACT FROM AUTHOR]

This paper proposes a method to modify high-amylose maize starch (HAMS) with lauric acid in an ethanol solvothermal system to prepare a cold-water swelling esterified starch (M-HAMS). Using M-HAMS as a granule stabilizer, oil-in-water Pickering emulsions were prepared, and factors affecting the oxidation stability of emulsion, such as granule content, oil-water ratio, pH value, and NaCl concentration, were studied. Atomic force microscopy (AFM) analysis showed that M-HAMS granules swell into dense molecular chains liking a three-dimensional network barrier in the emulsion, which increases the viscosity of continuous phase and prevents the oil droplets from contacting the pro-oxidant in the water phase, thus reducing the peroxide value. At the same time, the influence of changes in pH value and NaCl concentration on the viscosity and stability of emulsion is weakened. Therefore, the preparation of M-HAMS granule by ethanol solvothermal is a simple and effective method. [Display omitted] [ABSTRACT FROM AUTHOR]

Tick-borne Anaplasma species are obligate, intracellular, bacterial pathogens that cause important diseases globally in people, agricultural animals, and dogs. Targeted mutagenesis methods are yet to be developed to define genes essential for these pathogens. In addition, vaccines conferring protection against diseases caused by Anaplasma species are not available. Here, we describe a targeted mutagenesis method for deletion of the phage head-to-tail connector protein (phtcp) gene in Anaplasma marginale. The mutant did not cause disease and exhibited attenuated growth in its natural host (cattle). We then assessed its ability to confer protection against wild-type A. marginale infection challenge. Additionally, we compared vaccine protection with the mutant to that of whole cell A. marginale inactivated antigens as a vaccine (WCAV) candidate. Upon infection challenge, non-vaccinated control cattle developed severe disease, with an average 57% drop in packed cell volume (PCV) between days 26–31 post infection, an 11% peak in erythrocytic infection, and apparent anisocytosis. Conversely, following challenge, all animals receiving the live mutant did not develop clinical signs or anemia, or erythrocyte infection. In contrast, the WCAV vaccinees developed similar disease as the non-vaccinees following A. marginale infection, though the peak erythrocyte infection reduced to 6% and the PCV dropped 43%. This is the first study describing targeted mutagenesis and its application in determining in vivo virulence and vaccine development for an Anaplasma species pathogen. This study will pave the way for similar research in related Anaplasma pathogens impacting multiple hosts. Author summary: This is the first study describing targeted mutagenesis and its application in determining virulence and vaccine development for an Anaplasma species pathogen. The study is important in advancing similar research in related Anaplasma pathogens impacting multiple hosts. We demonstrated the successful development of a complete gene deletion mutation in A. marginale. Further, this in vivo infection study was performed in the natural host and established that growth of A. marginale is greatly limited when the phtcp gene is deleted. This study also demonstrated that cattle challenged with the virulent wild-type A. marginale four weeks after receiving the mutant A. marginale vaccine (MLAV) neither developed clinical disease nor did they exhibit hematologic abnormalities, such as anemia. On the contrary, non-vaccinated and WCAV vaccinated cattle challenged with virulent wild-type A. marginale developed clinical anaplasmosis with severe anemia, erythrocyte anisocytosis, and high-load infection in erythrocytes. [ABSTRACT FROM AUTHOR]

Plastic film mulching (PM) has been widely used to improve maize (Zea mays L.) yields and water use efficiency (WUE) in Northeast China, but the effects of PM in a changing climate characterized by highly variable precipitation are not well understood. Six site-year field experiments were conducted in the dry and rainy years to investigate the effects of PM on maize growth, grain yield, and WUE in Northeast China. Compared to crops grown without PM treatment (control, CK), PM significantly increased the grain yield by 15-26% in the dry years, but no significant yield increase was observed in the rainy years. Yield increase in the dry years was mainly due to a large increase in dry matter accumulation pre-silking compared to the CK, which resulted from a greater dry matter accumulation rate due to the higher topsoil temperature and water content. As a result, the WUE of the crops that underwent PM (3.27 kg m-3) treatment was also increased by around 16% compared to the CK, although the overall evapotranspiration was similar between the two treatments. In the rainy years, due to frequent precipitation and scant sunshine, the topsoil temperature and water content in the field that received PM treatment was improved only at some stages and failed to cause higher dry matter accumulation, except at the 8th leaf stage. Consequently, the grain yield and WUE were not improved by PM in the rainy years. In addition, we found that PM caused leaf senescence at the late growth stage in both dry and rainy years. Therefore, in practice, PM should be applied cautiously, especially when in-season precipitation is taken into account. [ABSTRACT FROM AUTHOR]

BACKGROUND Fruit and vegetables are frequently treated with fungicides to reduce possible spoilage. As a result, fungicide residues may be accumulated in derived products. This important group of chemical compounds has been heavily regulated because of their potential toxicity. Therefore, a simple and rapid method to determine fungicides is desired. RESULTS A simple non-aqueous capillary electrophoresis (NACE) method based on dispersive liquid-liquid microextraction (DLLME) has been proposed for the determination of imazalil, prochloraz and thiabendazole fungicides in fruits and juice samples. Separation buffer consisted of a methanol-acetonitrile mixture (35:65, v/v) containing 30 mmol L−1 ammonium chloride and 0.5% phosphoric acid. The optimum DLLME conditions were 80 µL trichloromethane as extraction solvent, 0.5 mL tetrahydrofuran as disperser solvent, sample solution pH at 6.0, 5% (w/v) NaCl and 10 s extraction time. Recoveries obtained for various samples ranged from 72% to 102%, with relative standard deviation lower than 6.4%. The limits of detection ranged from 0.47 to 0.72 µg kg−1. CONCLUSION The proposed method takes the advantages of DLLME and NACE. It is rapid, accurate, sensitive and reproducible for the determination of imazalil, prochloraz and thiabendazole in fruit samples. © 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Starch-based Pickering emulsions exhibit high interfacial stability in a certain range of mild pH environments. On the contrary, many studies have reported that when the pH value is <4, it often leads to different degrees of emulsion instability. In this paper, the microscopic state of starch granules in the emulsion and its effect on the stability of the emulsion were observed and analyzed by atomic force microscope (AFM) in tapping mode. At the same time, Pickering emulsions in acidic environment were prepared by using the gel properties of methyl cellulose (MC) in synergy with esterified high amylose maize starch (M-HAMS) granules. The results show that in the emulsion with pH 3, the excessive H + ion inhibits the swelling of M-HAMS granules and prevents it from forming a stable gel structure, which is the main cause of emulsion instability. The polarity of MC with water contact angle (WCA) of 81.8° is similar to that of M-HAMS granules with WCA of 80.1°, and a uniform and ordered micro-nanostructure is formed in the aqueous phase. The prepared acidic (pH 3–4) emulsion has good stability during the observation period of 30 days. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • The S-scheme CuFe 2 O 4 /ZnIn 2 S 4 heterojunction was successfully constructed. • The CuFe 2 O 4 /ZnIn 2 S 4 photocatalytic system exhibited a surprising production of H 2 O 2. • CuFe 2 O 4 /ZnIn 2 S 4 presented strong photocatalytic activity for the degradation of CIP. • The optimal pathway of H 2 O 2 generation and the degradation mechanism were confirmed. • The degradation pathway of CIP and toxicity analysis were further clarified. The photo-Fenton process is a promising technology for wastewater purification, but the extra addition of H 2 O 2 and the limited conversion of Fe2+/Fe3+ greatly hinder its practical application. In this study, we presented S-scheme CuFe 2 O 4 /ZnIn 2 S 4 heterojunction photocatalytic self-Fenton system for the degradation of ciprofloxacin (CIP). The formation of an internal electric field in CuFe 2 O 4 /ZnIn 2 S 4 heterojunction facilitated the transport and separation of photogenerated carriers, exhibiting high visible light absorption ability and photocatalytic activity. The CuFe 2 O 4 /ZnIn 2 S 4 photocatalytic system exhibited the surprisingly high H 2 O 2 selectivity (96.8 %) and H 2 O 2 yield (2545.4 µmol·g−1) through the 2e−-ORR process. DFT calculations found that the O 2 molecules were adsorbed on Cu atom of CuFe 2 O 4 /ZnIn 2 S 4 in a "Yeager-type" configuration, which facilitated the formation of a key intermediate (*OOH) for the conversion O 2 into H 2 O 2. The 25 mg·L−1 of CIP could be completely degraded within 60 min in CuFe 2 O 4 /ZnIn 2 S 4 photocatalytic system, while this system also had good cyclic stability and practicality. The photogenerated electrons that accumulated on conduction band of CuFe 2 O 4 facilitated the reduction of Fe3+ and Cu2+ to Fe2+ and Cu+, accompanying by the generation of OH, 1O 2 and O 2 −, while the photogenerated holes that accumulated on the valence band of ZnIn 2 S 4 could directly degrade pollutants. Finally, the vulnerable atomic sites of CIP were successfully predicted by Fukui function, while the CIP degradation pathway and toxicity analysis of degradation products were further clarified. This study provided a new design route for constructing photocatalytic self-Fenton system. [ABSTRACT FROM AUTHOR]

Rickettsiale diseases, including human monocytic ehrlichiosis caused by Ehrlichia chaffeensis, are the second leading cause of the tick-borne infections in the USA and a growing health concern. Little is known about how E. chaffeensis survives the host-induced stress in vertebrate and tick hosts. A molecular chaperone ClpB from several microorganisms has been reported to reactivate aggregated proteins in cooperation with the co-chaperones DnaK/DnaJ/GrpE (KJE). In this study, we performed the first biochemical characterization of ClpB from E. chaffeensis. The transcript of E. chaffeensis ClpB (EhClpB) is strongly upregulated after infection of cultured macrophages and its level remains high during the Ehrlichia replicative stage. EhClpB forms ATP-dependent oligomers and catalyzes the ATP hydrolysis, similar to E. coli ClpB (EcClpB), but its ATPase activity is insensitive to the EcClpB activators, casein and poly-lysine. EhClpB in the presence of E. coli KJE efficiently reactivates the aggregated glucose-6-phosphate dehydrogenase (G6PDH) and firefly luciferase. Unlike EcClpB, which requires the co-chaperones for aggregate reactivation, EhClpB reactivates G6PDH even in the absence of KJE. Moreover, EhClpB is functionally distinct from EcClpB as evidenced by its failure to rescue a temperature-sensitive phenotype of the clpB-null E. coli. The clpB expression pattern during the E. chaffeensis infection progression correlates with the pathogen’s replicating stage inside host cells and suggests an essential role of the disaggregase activity of ClpB in the pathogen’s response to the host-induced stress. This study sets the stage for assessing the importance of the chaperone activity of ClpB for E. chaffeensis growth within the mammalian and tick hosts. [ABSTRACT FROM AUTHOR]

Abstract: Environmental contaminants are frequently encountered as mixtures, and research on mixture toxicity is a hot topic until now. In the present study, the mixture toxicity of non-polar narcotic chemical was modeled by linear and nonlinear statistical methods, that is to say, by forward stepwise multilinear regression (MLR) and radial basis function neural networks (RBFNNs) from molecular descriptors that are calculated and be defined as composite descriptors according to the fractional concentrations of the mixture components. The statistical parameters provided by the MLR model were R 2 =0.9512, RMS=0.3792, F =1402.214 and LOOq 2 =0.9462 for the training set, and R 2 =0.9453, RMS=0.3458, F =276.671 and =0.9450 for the external test set. The RBFNN model gave the following statistical results, namely: R 2 =0.9779, RMS=0.2498, F =3188.202 and LOOq 2 =0.9746 for the training set, and R 2 =0.9763, RMS=0.2358, F =660.631 and =0.9745, for the external test set. Overall, these results suggest that the QSAR MLR-based model is a simple, reliable, credible and fast tool for the prediction mixture toxicity of non-polar narcotic chemicals. The RBFNN model gave even improved results. In addition, (the energy of the second lowest unoccupied molecular orbital) and PPSA (total charge weighted partial positively surface area) were found to have high correlation with the mixture toxicity. [Copyright &y& Elsevier]

Abstract: The maximum absorption wavelength (λ max) of a large data set of 191 azobenzene dyes was predicted by quantitative structure–property relationship (QSPR) tools. The λ max was correlated with the 4 molecular descriptors calculated from the structure of the dyes alone. The multiple linear regression method (MLR) and the non-linear radial basis function neural network (RBFNN) method were applied to develop the models. The statistical parameters provided by the MLR model were R 2 =0.893, , , F =1214.871, RMS =11.6430 for the training set; and R 2 =0.849, , , F =207.812, RMS =14.0919 for the external test set. The RBFNN model gave even improved statistical results: R 2 =0.920, , , F =1664.074, RMS =9.9215 for the training set, and R 2 =0.895, , , F =314.256, RMS =11.6427 for the external test set. This theoretical method provides a simple, precise and an alternative method to obtain λ max of azobenzene dyes. [Copyright &y& Elsevier]

Background: China is the birthplace of the deer family and the country with the most abundant deer resources. However, at present, China's deer industry faces the problem that pure sika deer and hybrid deer cannot be easily distinguished. Therefore, the development of a SNP identification chip is urgently required. Results: In this study, 250 sika deer, 206 red deer, 23 first-generation hybrid deer (F1), 20 s-generation hybrid deer (F2), and 20 third-generation hybrid deer (F3) were resequenced. Using the chromosome-level sika deer genome as the reference sequence, mutation detection was performed on all individuals, and a total of 130,306,923 SNP loci were generated. After quality control filtering was performed, the remaining 31,140,900 loci were confirmed. From molecular-level and morphological analyses, the sika deer reference population and the red deer reference population were established. The Fst values of all SNPs in the two reference populations were calculated. According to customized algorithms and strict screening principles, 1000 red deer-specific SNP sites were finally selected for chip design, and 63 hybrid individuals were determined to contain red deer-specific SNP loci. The results showed that the gene content of red deer gradually decreased in subsequent hybrid generations, and this decrease roughly conformed to the law of statistical genetics. Reaction probes were designed according to the screening sites. All candidate sites met the requirements of the Illumina chip scoring system. The average score was 0.99, and the MAF was in the range of 0.3277 to 0.3621. Furthermore, 266 deer (125 sika deer, 39 red deer, 56 F1, 29 F2,17 F3) were randomly selected for 1 K SNP chip verification. The results showed that among the 1000 SNP sites, 995 probes were synthesized, 4 of which could not be typed, while 973 loci were polymorphic. PCA, random forest and ADMIXTURE results showed that the 1 K sika deer SNP chip was able to clearly distinguish sika deer, red deer, and hybrid deer and that this 1 K SNP chip technology may provide technical support for the protection and utilization of pure sika deer species resources. Conclusion: We successfully developed a low-density identification chip that can quickly and accurately distinguish sika deer from their hybrid offspring, thereby providing technical support for the protection and utilization of pure sika deer germplasm resources. [ABSTRACT FROM AUTHOR]

A twin enrichment method based on offline dispersive liquid–liquid microextraction (DLLME) coupled with online field-amplified sample injection (FASI) was developed for the simultaneous determination of four sulfonamide (SA) antibiotics, including sulfamethazine (SMZ), sulfamerazine (SMR), sulfadizine (SDZ) and sulfacetamide (SFA), in different environmental waters, followed by capillary electrophoresis (CE). Various parameters that affected the separation performance of CE and the enrichment efficiencies of DLLME and FASI were optimized in detail, and excellent CE separation was attained within 6 min. The DLLME-FASI-CE offered high sensitivity enrichment factors of 206, 166, 185 and 150 for SMZ, SMR, SDZ and SFA, respectively. Highly sensitive detection was realized with low limits of detection (LODs), which ranged from 2.0–23.0, 2.2–26.0 and 4.3–63.0 ng mL−1 in tap water, lake water and seawater, respectively, as well as limits of quantification (LOQs) within 6.0–63.0, 7.4–96.0 and 14.0–201.0 ng mL−1, respectively. Satisfactory recoveries in the range of 91–108% were obtained with the three spiked environmental water samples, and the relative standard deviations were from 1.09–7.45%. The simple effective twin enrichment method provided promising perspective for CE determination of SAs in complicated aqueous matrices, with rapidity, sensitivity, and accuracy. [ABSTRACT FROM AUTHOR]

In this paper, the influence of surface oxide layer of SiC powder on the rheological behavior of its slurry was studied by acid washing, HF etching, and calcination oxidation processes. Zeta potential, particle size, inductively coupled plasma atomic emission spectrometry (ICP‐AES), energy dispersive X‐ray spectroscopy, and viscosity tests indicated the main factor that resulted in high viscosity of slurry is impure ions existing either on the powder surface or in the water phase, rather than the surface oxide layer. After the impure ions are removed, the surface oxide layer formed at a calcination temperature range of 400°C‐750°C does not cause an increase in the slurry viscosity, but contributes to an improvement in the rheological properties of SiC slurry. [ABSTRACT FROM AUTHOR]

Objective: We aimed to investigate the roles and underlying mechanisms of YAP in the proliferation of neuroblastoma cells. Methods: The expression level of YAP was evaluated by Western blotting and immunocytochemistry. Cell viability, cell proliferation and growth were detected by CCK‐8, PH3 and Ki67 immunostaining, and the real‐time cell analyser system. The nuclear and cytoplasmic proteins of p27Kip1 were dissociated by the nuclear‐cytosol extraction kit and were detected by Western blotting and immunocytochemistry. mRNA levels of Akt, CDK5 and CRM1 were determined by qRT‐PCR. Results: YAP was enriched in SH‐SY5Y cells (a human neuroblastoma cell line). Knock‐down of YAP in SH‐SY5Y cells or SK‐N‐SH cell line (another human neuroblastoma cell line) significantly decreased cell viability, inhibited cell proliferation and growth. Mechanistically, knock‐down of YAP increased the nuclear location of p27Kip1, whereas serum‐induced YAP activation decreased the nuclear location of p27Kip1 and was required for cell proliferation. Meanwhile, overexpression of YAP in these serum‐starved SH‐SY5Y cells decreased the nuclear location of p27Kip1, promoted cell proliferation and overexpression of p27Kip1 in YAP‐activated cells inhibited cell proliferation. Furthermore, knock‐down of YAP reduced Akt mRNA and protein levels. Overexpression of Akt in YAP‐downregulated cells decreased the nuclear location of p27Kip1 and accelerated the proliferation of SH‐SY5Y cells. Conclusions: Our studies suggest that YAP promotes the proliferation of neuroblastoma cells through negatively controlling the nuclear location of p27Kip1 mediated by Akt. [ABSTRACT FROM AUTHOR]

It is very important for the polyimide film that a dense and uniform SiO x coating on its surface to resist atomic oxygen (AO) erosion. In this paper, an improved plasma polymerization process is employed to prepare a SiO x protective coating on the surface of Kapton. A deposition process is transited gradually from organic layer to inorganic layer by varying the ratio of O 2 /Ar and a uniform and compact organic-inorganic hybrid coating is easily formed. The samples are tested by AO irradiation in a ground-based simulator. When the total AO fluence is up to 1.09 × 1022 atoms/cm2, the uncoated sample has already been eroded and broken completely. On the contrary, though the color of coated sample has turned dark, but the sample still keeps intact. The AO erosion yield of coated sample is sharply down, equivalent to only 6.0% of pristine Kapton. • The SiOx coating was gradually grown from organic to inorganic layer through plasma polymerization. • The adhesion between the SiOx coating and Kapton substrate can be enhanced by the gradient deposition process. • After atomic oxygen exposure 1.09 × 1022 atom/cm2, the coated Kapton kept perfectly. [ABSTRACT FROM AUTHOR]

Polyhedral oligomeric silsesquioxane (POSS) nanoparticles have attracted wide attention due to their unique capability to reinforce polymers. However, if POSS cannot be uniformly dispersed in the polymer matrix, the desired properties of composites will be reduced. To solve this problem, a method of in situ formation of POSS layer on the surface of polyimide (Kapton) was explored, which not only kept the original properties of polymer unaffected, but also endowed the composites with new properties. The Kapton film completely wetted by γ-aminopropyltriethoxysilane (APTES) was placed in a solution containing tetraethylammonium hydroxide (TEAOH) catalyst, and a POSS layer was gradually grown on the Kapton surface. Subsequently, SiO 2 coating was deposited on the surface of POSS layer by magnetron sputtering to prepare SiO 2 /POSS/Kapton (SPK) samples. FTIR and XPS analyses showed that a layer with POSS structure was formed on the surface of Kapton after 6 h self-growth. Atomic oxygen irradiation test shows that the erosion rate of 6 h SPK sample is 0.19 × 10−24 cm3/atom, only equivalent to 6.0 % of original Kapton, which improves the corrosion resistance of polymer materials. [Display omitted] • Solvothermal treatment of toluene/APTES is beneficial to an in situ formation of POSS layer on the surface of Kapton. • Compared with bulk doping, the original properties of Kapton films can be perfectly maintained by in situ formation of POSS. • As a transition layer, POSS provides a new idea to solve the problem of cracking and spalling of SiO 2 coating. [ABSTRACT FROM AUTHOR]

In this paper, the esterification modification of different kinds of starches such as waxy maize, normal maize, high-amylose maize, cassava and potato in high temperature closed system were studied by solvothermal method. The oil-in-water Pickering emulsion were prepared with esterified starches as granule stabilizer. The microscopic state of granules in the emulsion and the physical and oxidation stability of emulsion were studied. The results show that starches are not gelatinized and can be esterified at a temperature (100 °C) much higher than that of gelatinization, and the granule morphology is almost unchanged. DS (degree of substitution) values of esterified starches range from 0.0333 to 0.0512. Pickering emulsion with 50 vol% oil volume fraction prepared with 3.0 wt% granule concentration did not show any instability such as oil-water separation after storage at room temperature for 30 days. Atomic force microscope (AFM) analysis showed that all esterified starch granules had the characteristics of granular cold-water swelling starch (GCWSS). The granules completely swelled into a dense molecular chain in the emulsion, and this three-dimensional network structure improved the stability of emulsion. Therefore, the preparation of esterified starch granules by ethanol solvothermal method is a simple and effective method. [Display omitted] [ABSTRACT FROM AUTHOR]

Cardiac troponin I (cTnI) was considered as the “gold standard” for acute myocardial infarction (AMI) diagnosis owing to its superior cardiac specificity for cardiac damage and showing little or no changes in patients with a skeletal muscle disease or trauma. Herein, a new signal amplification surface-enhanced Raman scattering (SERS) platform was developed for recognition and detection of cTnI by using gold nanoparticles (AuNPs), graphene oxide (GO) and magnetic beads (MB). Here, antibody/Raman reporter labeled AuNP–functionalized GO were employed as both SERS nanotags and signal amplification carriers. Monoclonal antibody modified MB were applied as the capture probe and separation agents. In the presence of cTnI, sandwich type immunocomplexes, “capture probe/target/SERS nanotags”, were formed through antibody–antigen–antibody interactions. Due to the strong SERS enhancement ability of the designed GO/AuNP complexes and a high binding chance between cTnI and the GO/AuNP complexes, the proposed SERS-based immunoassay could selectively detect cTnI with a high sensitivity (detection limit of 5 pg mL−1) and a good linearity was obtained in a range of 0.01–1000 ng mL−1. In addition, this method was also successfully applied for detecting cTnI in serum substitute media with a similar linear range. Furthermore, this strategy can be constructed with different kinds of antibodies and Raman reporters, and thus can be easily used for simultaneous detection of multiple biomarkers. Therefore, this proposed SERS-based signal amplification technique shows strong potential for the clinical diagnosis of AMI disease. [ABSTRACT FROM AUTHOR]

Abstract Olfactory ensheathing cells (OECs) migrate from olfactory epithelium towards olfactory bulb (OB), contributing to formation of the presumptive olfactory nerve layer during development. However, it remains unclear that molecular mechanism of regulation of OEC migration in OB. In the present study, we found that OECs highly expressed the receptors of semaphorin 3A (Sema3A) in vitro and in vivo , whereas Sema3A displayed a gradient expression pattern with higher in inner layer of OB and lower in outer layer of OB. Furthermore, the collapse assays, Boyden chamber migration assays and single-cell migration assays showed that Sema3A induced the collapse of leading front of OECs and inhibited OEC migration. Thirdly, the leading front of OECs exhibited adaptation in a protein synthesis-independent manner, and endocytosis-dependent manner during Sema3A-induced OEC migration. Finally, Sema3A-induced collapse of leading front was required the decrease of focal adhesion and a retrograde F-actin flow in a cofilin activation-dependent manner. Taken together, these results demonstrate that Sema3A as an inhibitive migratory factor for OEC migration through cofilin activation is involved in the formation of olfactory nerve layer. Highlights • Sema3A induces the collapse of OECs and inhibited OEC migration. • OECs exhibit adaptation induced by Sema3A in an endocytosis-dependent manner. • A retrograde F-actin flow by cofilin activation is required for Sema3A-induced collapse in OECs. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome (mtDNA) of Peking duck (Anas platyrhychos) was determined. The entire genome was 16,604 bp in length. Similar to the typical mtDNA of vertebrates, it contained 37 genes (13 protein-coding genes, 2 rRNA genes, 22 tRNA genes) and a non-coding region (D-loop). The characteristics of the mitochondrial genome was analyzed and discussed in detail. [ABSTRACT FROM AUTHOR]