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The development of a tuberculosis (TB) vaccine is imperative. Employing multi-stage Mycobacterium tuberculosis (Mtb) antigens as targeted antigens represents a critical strategy in establishing an effective novel TB vaccine. In this investigation, we evaluated the immunogenicity and protective efficacy of a recombinant adenovirus vaccine expressing two fusion proteins, Ag85B-ESAT6 (AE) and Rv2031c-Rv2626c (R2), derived from multi-stage antigens of Mtb via intranasal administration in mice. Intranasal delivery of Ad-AE-R2 induced both long-lasting mucosal and systemic immunities, with a preferential elicitation of CD8+ T cell immunity demonstrated by the accumulation and retention of CD8+ T cells in BALF, lung, and spleen, as well as the generation of CD8+ TRM cells in BALF and lung tissues. Compared to subcutaneous immunization with Bacillus Calmette-Guerin (BCG), Ad-AE-R2 provided superior protection against high-dose intratracheal BCG challenge, specifically within the lungs of mice. Our findings support the notion that empowering T cells within the respiratory mucosa is crucial for TB vaccine development while highlighting targeting CD8+ T cell immunity as an effective strategy for optimizing TB vaccines and emphasizing that eliciting systemic memory immunity is also vital for the successful development of a TB mucosal vaccine. Furthermore, our results demonstrate that the BCG challenge serves as a convenient and efficient method to evaluate candidate vaccine efficacy. [ABSTRACT FROM AUTHOR]

In this study, "Honghu White Lotus", "Red Lotus (HH)", "Hunan Cunshan Lotus (CS)", "Wuyi Xuanlian", "Space Lotus 36", "Fujian Jianning White Lotus (JB)", "Jiangsu Yangzhou Lotus (JY)", and "Suzhou Dongshan Lotus" were selected as experimental subjects. The lotus seed flesh and lotus plumule of each cultivar were selected for nutritional quality and functional active substance analyses. Comparing different cultivars of lotus seeds, the protein and crude fat contents of JY flesh were the highest at 65.59 mg/g and 13%, respectively. The VC content of JB flesh and lotus plumule is the highest at 41.56 mg/g and 204.29 mg/g, respectively. JB flesh has the lowest soluble sugar content, at 17.87 mg/g, while HB's lotus plumule and flesh have the highest content, at 33.67 mg/g and 29.62 mg/g, respectively. There was no significant difference in the crude fat content of the flesh and lotus plumule among the eight cultivars. TK flesh and lotus plumule have the highest amylose content, at 23.67 mg/g and 76.81 mg/g, respectively. Among them, the total starch content of JB (476.17 mg/g) was relatively high, whereas its amylose content was only 26.09 mg/g. Lower amylose content makes it less prone to aging. The total phenolic and flavonoid contents of the JY lotus plumule were the highest, at 18.64 and 21.04 mg/g, respectively. The alkaloid content of CS, HH, and JY was relatively high at 20.01, 19.29, and 18.68 mg/g, respectively. These can provide a consultation for the estimation and processing of the nutritional quality of different lotus seeds. [ABSTRACT FROM AUTHOR]

This study investigated the impact of variety and harvest time on the visual appearance, nutritional quality, and functional active substances of six lotus root cultivars: "Xinsanwu", "Wuzhi No. 2", "Baiyuzhan", "Huaqilian", "Elian No. 6", and "Elian No. 5". Samples were collected monthly from December 2023 to April 2024. A nutrient analysis revealed a decrease in the water content with a delayed harvest. The total soluble solids and soluble sugar content peaked towards the end and middle-to-late harvest periods, respectively. Starch levels initially increased before declining, while the soluble protein exhibited a triphasic trend with an initial rise, a dip, and a final increase. The vitamin C (Vc) content varied across cultivars. Functional active substances displayed dynamic changes. The total phenolics initially decreased, then increased, before ultimately declining again. The total flavonoid content varied by both cultivar and harvest time. The phenolic acid and flavonoid content mirrored the trends observed for total phenolics and total flavonoids. Gastrodin was the most abundant non-flavonoid compound across all varieties. "Wuzhi No. 2" and "Baiyuzhan" displayed higher levels of functional active substances and starch, while the Elian series and "Xinsanwu" cultivar exhibited a greater content of Vc, soluble sugar, and soluble protein. Specific harvest periods yielded optimal results: "Wuzhi No. 2" (H1 and H5), "Huaqilian" (H2), and "Baiyuzhan" (H3 and H4) demonstrated a high nutrient and functional active substance content. Overall, the lotus roots harvested in period H4 achieved the highest score. Overall, this study provides the foothold for the rapid identification of superior lotus root cultivars and the valorization of lotus root by-products via advanced processing methods. Additionally, it offers valuable insights for market participants and consumers to select optimal varieties and harvest times based on their specific needs. [ABSTRACT FROM AUTHOR]

Mica is a harmful substance in sand and occurs frequently. The application of super standard mica sand is a difficult problem in large-scale engineering. In this work, the effects of an interface modifier, mineral admixture, and a curing system on the properties of cement-based materials with super standard mica sand were studied. The strength of cement-based materials linearly decreases with the mica content in sand. When the mica content in sand exceeds 6%, the compressive strength of mortar and concrete at 28 d decreases by more than 22.3% and 33.5%, respectively. By adding the silane coupling agent (SCA) of 50% mica mass and curing in natural conditions, the compressive strength of mortar increases by 10.9%. The cement-based materials with the SCA are more suitable for curing in natural conditions, and the performance of the SCA will not be affected by adding appropriate amounts of mineral admixture. The drying shrinkage strain of the concrete, with the sand containing high mica content modified by SCA, is reduced by 10.5%, and the diffusion of chloride ions in concrete is reduced. The XRD results show that the addition of the interfacial agent does not change the hydration products. The MIP and SEM results show that the SCA can form a bridge structure between the hydration products and the mica, improve the bonding strength of the interface zone, and reduce the number of harmful pores. [ABSTRACT FROM AUTHOR]

The 1092 bp F3H gene from Trapa bispinosa Roxb., which was named TbF3H, was cloned and it encodes 363 amino acids. Bioinformatic and phylogenetic tree analyses revealed the high homology of TbF3H with flavanone 3-hydroxylase from other plants. A functional analysis showed that TbF3H of Trapa bispinosa Roxb. encoded a functional flavanone 3-hydroxylase; it catalyzed the formation of dihydrokaempferol (DHK) from naringenin in S. cerevisiae. The promoter strengths were compared by fluorescence microscopy and flow cytometry detection of the fluorescence intensity of the reporter genes initiated by each constitutive promoter (FITC), and DHK production reached 216.7 mg/L by the promoter adjustment strategy and the optimization of fermentation conditions. The results presented in this study will contribute to elucidating DHK biosynthesis in Trapa bispinosa Roxb. [ABSTRACT FROM AUTHOR]

Background: Trapa bispinosa shells (TBs) and its flesh (TBf) have been recognized for their medicinal properties, including antioxidant, antitumor, and immunomodulatory effects. Despite these benefits, TBs are often discarded as waste material, and their applications remain to be further explored. Methods: In this study, we optimized the solid-state fermentation process of Ganoderma sinense (GS) with TBs using a response surface experiment methodology to obtain the fermented production with the highest water extract rate and DPPH free radical scavenging activity. We prepared and characterized pre-fermentation purified polysaccharides (P1) and post-fermentation purified polysaccharides (P2). Alcoholic extracts before (AE1) and after (AE2) fermentation were analyzed for active components such as polyphenols and flavonoids using UPLC-QTOF-MS/MS (ultra-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry). Mouse macrophages (RAW 264.7) were employed to compare the immune-stimulating ability of polysaccharides and the antioxidant activity of AE1 and AE2. Results: Optimal fermentation conditions comprised a duration of 2 days, a temperature of 14 °C, and a humidity of 77%. The peak water extract yield and DPPH free radical scavenging rate of the water extract from TBs fermented by GS were observed under these conditions. The enhanced activity may be attributed to changes in the polysaccharide structure and the components of the alcoholic extract. The P2 treatment group indicated more secretion of RAW 264.7 cells of NO, iNOS, IL-2, IL-10, and TNF-α than P1, which shows that the polysaccharides demonstrated increased immune-stimulating ability, with their effect linked to the NF-кB pathway. Moreover, the results of the AE2 treatment group indicated that secretion of RAW 264.7 cells of T-AOC and T-SOD increased and MDA decreased, which shows that the alcoholic extract demonstrated enhanced antioxidant activity, with its effect linked to the Nrf2/Keap1-ARE pathway. Conclusions: Biphasic fermentation of Trapa bispinosa shells by Ganoderma sinense could change the composition and structure of the polysaccharides and the composition of the alcoholic extract, which could increase the products' immunomodulatory and antioxidant activity. [ABSTRACT FROM AUTHOR]

Aberrantly high dietary cholesterol intake and intestinal cholesterol uptake lead to dyslipidemia, one of the risk factors for cardiovascular diseases (CVDs). Based on previous studies, laminarin, a polysaccharide found in brown algae, has hypolipidemic activity, but its underlying mechanism has not been elucidated. In this study, we investigated the effect of laminarin on intestinal cholesterol uptake in vitro, as well as the lipid and morphological parameters in an in vivo model of high-fat diet (HFD)-fed mice, and addressed the question of whether Niemann–Pick C1-like 1 protein (NPC1L1), a key transporter mediating dietary cholesterol uptake, is involved in the mechanistic action of laminarin. In in vitro studies, BODIPY-cholesterol-labeled Caco-2 cells were examined using confocal microscopy and a fluorescence reader. The results demonstrated that laminarin inhibited cholesterol uptake into Caco-2 cells in a concentration-dependent manner (EC50 = 20.69 μM). In HFD-fed C57BL/6J mice, laminarin significantly reduced the serum levels of total cholesterol (TC), total triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C). It also decreased hepatic levels of TC, TG, and total bile acids (TBA) while promoting the excretion of fecal cholesterol. Furthermore, laminarin significantly reduced local villous damage in the jejunum of HFD mice. Mechanistic studies revealed that laminarin significantly downregulated NPC1L1 protein expression in the jejunum of HFD-fed mice. The siRNA-mediated knockdown of NPC1L1 attenuated the laminarin-mediated inhibition of cholesterol uptake in Caco-2 cells. This study suggests that laminarin significantly improves dyslipidemia in HFD-fed mice, likely by reducing cholesterol uptake through a mechanism that involves the downregulation of NPC1L1 expression. [ABSTRACT FROM AUTHOR]

To confirm the effect of confining pressure on the dynamic mechanical behavior of ultrahigh-performance concrete (UHPC), this study used a true triaxial split Hopkinson pressure bar test system to perform dynamic compression tests on UHPC under triaxial constraints. The confining pressure range considered was 5~10 MPa, the strain rate range was 35~80 s−1, and the steel fiber contents were 0.5%, 1% and 2%. The three-dimensional dynamic engineering stress-strain relationship and equivalent stress-strain relationship of UHPC under different confining pressures and different strain rates were obtained and analyzed in detail. The results show that under the confinement condition, the dynamic peak axial stress–strain and dynamic peak lateral stress–strain of UHPC have strong sensitivity to the strain rate. In addition, the dynamic peak lateral stress–strain is more sensitive to the confining pressure than the dynamic axial stress. An empirical strength enhancement factor (DIFc) that considers the strain rate effect and confining pressure was derived, and the impact of the coupling between the enhancement caused by the confining pressure and the strain rate effect on the dynamic strength of the UHPC under triaxial confinement was discussed. A dynamic strength failure criterion for UHPC under triaxial constraint conditions was established. [ABSTRACT FROM AUTHOR]

Allium mongolicum Regel (A. mongolicum) is a healthy edible plant but highly perishable with a short shelf life of 1–2 d. Modified atmosphere packaging (MAP) could inhibit the postharvest senescence and decay of the vegetables. Thus, the aim of this study was to apply MAP with different gas permeabilities to the storage of A. mongolicum and evaluate its effects on maintaining microbial communities and the postharvest quality of A. mongolicum. The results showed that polypropylene/poly(butylene adipate-co-terephthalate) (PP/PBAT, abbreviated as PAT) MAP was suitable for the storage of A. mongolicum by establishing an optimal atmosphere of 0.5–0.6% O2 and 6.2–7.1% CO2 in the bag. It could delay the postharvest senescence of A. mongolicum and maintain its quality by slowing down its respiration rate and weight loss, reducing cell membrane permeability and lipid peroxidation, maintaining the cell wall, and reducing infection and the growth of microorganisms. However, A. mongolicum in HPT was more perishable than that in PAT during storage. Pseudomonas was found to be the main spoilage bacteria, and they could also be effectively inhibited by PAT-MAP. The next-generation sequencing results also showed the growth of Escherichia-Shigella, Clostridium sensu stricto 1, Streptococcus, Aureobasidium, Didymella, and Fusarium, responsible for A. mongolicum decay or human disease, was well inhibited by PAT-MAP. The results suggested that PAT-MAP could be used to maintain microbial diversity and the postharvest quality of A. mongolicum under cold storage conditions. It provided a feasible solution for the preservation, food quality, and safety control of A. mongolicum. [ABSTRACT FROM AUTHOR]

We present a novel fast synthetic aperture radar (SAR) image simulation method based on the echo matrix cell algorithm including multiple scattering. To improve the efficiency of SAR image simulation while ensuring the fidelity of the simulated results, we first discretized the target facets set in the SAR beams footprint into lattice targets using the range-Doppler (RD) imaging geometry model and provided the basis for simulating electromagnetic wave transmission. Based on the simulation of electromagnetic waves transmission, we used the ray tracing algorithm to calculate the multiple backscattering field including multi-polarimetric information and various material properties. Then, based on the echo matrix cell algorithm, we set the echo matrix cell as the subfield of the target backscattering field and designed the CUDA kernel function to implement a computation parallelization for SAR echo generation. All the echo matrix cells are traversed in parallel to obtain the total backscattering field of the target, reproducing the time-varying characteristic of the backscatter coefficient for each lattice target within the synthetic aperture time. The echo signal is processed using the RD imaging algorithm to obtain the simulated SAR image. Finally, we select some targets including aircraft carrier and airplane models for simulation tests. The computation efficiency is improved over 170-fold by comparing the computations of the proposed method and CPU single-thread. We also performed some qualitative and quantitative evaluations on the fidelity of the simulated SAR images. The experimental results verify the effectiveness of the proposed method. [ABSTRACT FROM AUTHOR]

Waves of breakthrough infections by SARS-CoV-2 Omicron subvariants currently pose a global challenge to the control of the COVID-19 pandemic. We previously reported a pVAX1-based DNA vaccine candidate, pAD1002, that encodes a receptor-binding domain (RBD) chimera of SARS-CoV-1 and Omicron BA.1. In mouse and rabbit models, pAD1002 plasmid induced cross-neutralizing Abs against heterologous sarbecoviruses, including SARS-CoV-1 and SARS-CoV-2 wildtype, Delta and Omicron variants. However, these antisera failed to block the recent emerging Omicron subvariants BF.7 and BQ.1. To solve this problem, we replaced the BA.1 RBD-encoding DNA sequence in pAD1002 with that of BA.4/5. The resulting construct, namely pAD1016, elicited SARS-CoV-1 and SARS-CoV-2 RBD-specific IFN-γ+ cellular responses in BALB/c and C57BL/6 mice. More importantly, pAD1016 vaccination in mice, rabbits and pigs generated serum Abs capable of neutralizing pseudoviruses representing multiple SARS-CoV-2 Omicron subvariants including BA.2, BA.4/5, BF.7, BQ.1 and XBB. As a booster vaccine for inactivated SARS-CoV-2 virus preimmunization in mice, pAD1016 broadened the serum Ab neutralization spectrum to cover the Omicron BA.4/5, BF7 and BQ.1 subvariants. These preliminary data highlight the potential benefit of pAD1016 in eliciting neutralizing Abs against broad-spectrum Omicron subvariants in individuals previously vaccinated with inactivated prototype SARS-CoV-2 virus and suggests that pAD1016 is worthy of further translational study as a COVID-19 vaccine candidate. [ABSTRACT FROM AUTHOR]

(1) Background: Euryale ferox Salisb is a large aquatic plant of the water lily family and an edible economic crop with medicinal value. The annual output of Euryale ferox Salisb shell in China is higher than 1000 tons, often as waste or used as fuel, resulting in waste of resources and environmental pollution. We isolated and identified the corilagin monomer from Euryale ferox Salisb shell and discovered its potential anti-inflammatory effects. This study aimed to investigate the anti-inflammatory effect of corilagin isolated from Euryale ferox Salisb shell. (2) Methods: We predict the anti-inflammatory mechanism by pharmacology. LPS was added to 264.7 cell medium to induce an inflammatory state, and the safe action range of corilagin was screened using CCK-8. The Griess method was used to determine NO content. The presence of TNF-α, IL-6, IL-1β, and IL-10 was determined by ELISA to evaluate the effect of corilagin on the secretion of inflammatory factors, while that of reactive oxygen species was detected by flow cytometry. The gene expression levels of TNF-α, IL-6, COX-2, and iNOS were determined using qRT-PCR. qRT-PCR and Western blot were used to detect the mRNA and expression of target genes in the network pharmacologic prediction pathway. (3) Results: Network pharmacology analysis revealed that the anti-inflammatory effect of corilagin may be related to MAPK and TOLL-like receptor signaling pathways. The results demonstrated the presence of an anti-inflammatory effect, as indicated by the reduction in the level of NO, TNF-α, IL-6, IL-1β, IL-10, and ROS in Raw264.7 cells induced by LPS. The results suggest that corilagin reduced the expression of TNF-α, IL-6, COX-2, and iNOS genes in Raw264.7 cells induced by LPS. The downregulation of the phosphorylation of IκB-α protein related to the toll-like receptor signaling pathway and upregulation of the phosphorylation of key proteins in the MAPK signaling pathway, P65 and JNK, resulted in reduced tolerance toward lipopolysaccharide, allowing for the exertion of the immune response. (4) Conclusions: The results demonstrate the significant anti-inflammatory effect of corilagin from Euryale ferox Salisb shell. This compound regulates the tolerance state of macrophages toward lipopolysaccharide through the NF-κB signaling pathway and plays an immunoregulatory role. The compound also regulates the expression of iNOS through the MAPK signaling pathway, thereby alleviating the cell damage caused by excessive NO release. [ABSTRACT FROM AUTHOR]

The SARS-CoV-2 pandemic remains an ongoing threat to global health with emerging variants, especially the Omicron variant and its sub-lineages. Although large-scale vaccination worldwide has delivered outstanding achievements for COVID-19 prevention, a declining effectiveness to a different extent in emerging SARS-CoV-2 variants was observed in the vaccinated population. Vaccines eliciting broader spectrum neutralizing antibodies and cellular immune responses are urgently needed and important. To achieve this goal, rational vaccine design, including antigen modeling, screening and combination, vaccine pipelines, and delivery, are keys to developing a next-generation COVID-19 vaccine. In this study, we designed several DNA constructs based on codon-optimized spike coding regions of several SARS-CoV-2 variants and analyzed their cross-reactive antibodies, including neutralizing antibodies, and cellular immune responses against several VOCs in C57BL/6 mice. The results revealed that different SARS-CoV-2 VOCs induced different cross-reactivity; pBeta, a DNA vaccine encoding the spike protein of the Beta variant, elicited broader cross-reactive neutralizing antibodies against other variants including the Omicron variants BA.1 and BA.4/5. This result demonstrates that the spike antigen from the Beta variant potentially serves as one of the antigens for multivalent vaccine design and development against variants of SARS-CoV-2. [ABSTRACT FROM AUTHOR]

Trapa bispinosa Roxb. is an economical crop for medicine and food. Its roots, stems, leaves, and pulp have medicinal applications, and its shell is rich in active ingredients and is considered to have a high medicinal value. One of the main functional components of the Trapa bispinosa Roxb. shell is 1-galloyl-beta-D-glucose (βG), which can be used in medical treatment and is also an essential substrate for synthesizing the anticancer drug beta-penta-o-Galloyl-glucosen (PGG). Furthermore, gallate 1-beta-glucosyltransferase (EC 2.4.1.136) has been found to catalyze gallic acid (GA) and uridine diphosphate glucose (UDPG) to synthesize βG. In our previous study, significant differences in βG content were observed in different tissues of Trapa bispinosa Roxb. In this study, Trapa bispinosa Roxb. was used to clone 1500 bp of the UGGT gene, which was named TbUGGT, to encode 499 amino acids. According to the specificity of the endogenous expression of foreign genes in Escherichia coli, the adaptation codon of the cloned original genes was optimized for improved expression. Bioinformatic and phylogenetic tree analyses revealed the high homology of TbUGGT with squalene synthases from other plants. The TbUGGT gene was constructed into a PET-28a expression vector and then transferred into Escherichia coli Transsetta (DE3) for expression. The recombinant protein had a molecular weight of 55 kDa and was detected using SDS-PAGE. The proteins were purified using multiple fermentation cultures to simulate the intracellular environment, and a substrate was added for in vitro reaction. After the enzymatic reaction, the levels of βG in the product were analyzed using HPLC and LC-MS, indicating the catalytic activity of TbUGGT. The cloning and functional analysis of TbUGGT may lay the foundation for further study on the complete synthesis of βG in E. coli. [ABSTRACT FROM AUTHOR]

Natural antioxidants derived from agricultural by-products have great promise and ecological advantages in the treatment of oxidative stress-related diseases. The eggshell membrane (ESM) from hatched eggs, i.e., the hatched ESM, is a globally abundant agricultural byproduct, and its high-value utilization has been rarely studied compared to the well-studied ESM from fresh eggs. In this research, we systematically characterized the hatched ESM as a novel source of antioxidant hydrolysates and explored their potential role in H2O2-induced human chondrocytes. The results showed that the hatched ESM is a protein-rich fibrous mesh material with a significantly different structure and composition from those of fresh ESM. Enzymatic hydrolysis of hatched ESM can produce antioxidant hydrolysates rich in low molecular weight (MW) peptides, which mainly derived from the Lysyl oxidase homolog by Nano-LC-MS/MS analysis. The peptide fraction with MW < 3 kDa (HEMH-I) exhibited the highest DPPH radical scavenging, Fe2+-chelating, and Fe3+-reducing abilities. In H2O2-induced human SW1353 chondrocytes, HEMH-I treatment significantly increased the cell viability and ameliorated oxidative stress, inflammatory response, and cartilage matrix degradation by reducing the level of ROS, matrix metalloprotease 3 (MMP3), MMP13, and IL-6, and by promoting the expression of SOD and type II collagen, potentially through activating the cellular Keap1/Nrf2/HO-1 pathway. This study provides a theoretical basis for the value-added application of hatched ESM waste to produce antioxidant hydrolysates and indicates their potential as functional food and pharmaceuticals. [ABSTRACT FROM AUTHOR]

We present a novel echo-based synthetic aperture radar (SAR) image simulation method that comprehensively utilizes both SAR effective view and ray tracing algorithms. To improve the fidelity of simulated SAR images, we first design an SAR effective view algorithm to process the selected facet target model, with the purpose of discretizing the facets in the SAR effective view into lattice targets and ensuring that the interval of lattice targets is set strictly following the Nyquist sampling law. Then, we combine the ray tracing algorithm and SAR echo time-domain simulation and perform SAR echo time-domain simulation of lattice targets based on ray tracing. Various kinds of backscatter coefficient for each point target can be recorded, corresponding to the number of transmitted pulses within the synthetic aperture time. The echo matrixes of lattice targets are superimposed to generate the raw echo signal. Finally, the raw echo signal is processed by using the range-Doppler (RD) imaging algorithm to obtain the simulated SAR image. We conducted SAR image simulation tests on several facet target models, including car body, assault boat, and airplane, with different material properties. The simulated SAR images obtained by the proposed method were qualitatively and quantitatively evaluated. The experimental results verify the effectiveness of the proposed method. [ABSTRACT FROM AUTHOR]

Artemisiae argyi Folium is a traditional herbal medicine used for moxibustion heat therapy in China. The volatile oils in A.argyi leaves are closely related to its medicinal value. Records suggest that the levels of these terpenoids components within the leaves vary as a function of harvest time, with June being the optimal time for A. argyi harvesting, owing to the high levels of active ingredients during this month. However, the molecular mechanisms governing terpenoid biosynthesis and the time-dependent changes in this activity remain unclear. In this study, GC–MS analysis revealed that volatile oil levels varied across four different harvest months (April, May, June, and July) in A. argyi leaves, and the primarily terpenoids components (including both monoterpenes and sesquiterpenes) reached peak levels in early June. Through single-molecule real-time (SMRT) sequencing, corrected by Illumina RNA-sequencing (RNA-Seq), 44 full-length transcripts potentially involved in terpenoid biosynthesis were identified in this study. Differentially expressed genes (DEGs) exhibiting time-dependent expression patterns were divided into 12 coexpression clusters. Integrated chemical and transcriptomic analyses revealed distinct time-specific transcriptomic patterns associated with terpenoid biosynthesis. Subsequent hierarchical clustering and correlation analyses ultimately identified six transcripts that were closely linked to the production of these two types of terpenoid within A. argyi leaves, revealing that the structural diversity of terpenoid is related to the generation of the diverse terpene skeletons by prenyltransferase (TPS) family of enzymes. These findings can guide further studies of the molecular mechanisms underlying the quality of A. argyi leaves, aiding in the selection of optimal timing for harvests of A. argyi. [ABSTRACT FROM AUTHOR]

Although winter wheat has been mapped by remote sensing in several studies, such mapping efforts did not sufficiently utilize contextual information to reduce the noise and still depended heavily on optical imagery and exhausting classification approaches. Furthermore, the influence of similarity measures on winter wheat identification remains unclear. To overcome these limitations, this study developed an object-based automatic approach to map winter wheat using multitemporal Sentinel-1 (S1) and Sentinel-2 (S2) imagery. First, after S1 and S2 images were preprocessed, the Simple Non-Iterative Clustering (SNIC) algorithm was used to conduct image segmentation to obtain homogeneous spatial objects with a fusion of S1 and S2 bands. Second, the temporal phenology patterns (TPP) of winter wheat and other typical land covers were derived from object-level S1 and S2 imagery based on the collected ground truth samples, and two improved distance measures (i.e., a composite of Euclidean distance and Spectral Angle Distance, (ESD) and the difference–similarity factor distance (DSF)) were built to evaluate the similarity between two TPPs. Third, winter wheat objects were automatically identified from the segmented spatial objects by the maximum between-class variance method (OTSU) with distance measures based on the unique TPP of winter wheat. According to ground truth data, the DSF measure was superior to other distance measures in winter wheat mapping, since it achieved the best overall accuracy (OA), best kappa coefficient (Kappa) and more spatial details for each feasible band (i.e., NDVI, VV, and VH/VV), or it obtained results comparable to those for the best one (e.g., NDVI + VV). The resultant winter wheat maps derived from the NDVI band with the DSF measure achieved the best accuracy and more details, and had an average OA and Kappa of 92% and 84%, respectively. The VV polarization with the DSF measure produced the second best winter wheat maps with an average OA and Kappa of 91% and 80%, respectively. The results indicate the great potential of the proposed object-based approach for automatic winter wheat mapping for both optical and Synthetic Aperture Radar (SAR) imagery. [ABSTRACT FROM AUTHOR]

Cellulases play key roles in the degradation of lignocellulosic materials. The function and mechanism of the catalytic domain (CD) and carbohydrate-binding module (CBM) of cellulases were earlier revealed by analysis and characterization of protein structure. However, understanding of the catalytic mechanism of the entire enzyme, and the analysis of the catalytic model, were inadequate. Therefore, the linker chain between CD and CBM has been extensively studied to bridge this gap. Cellulase AcCel12B and three mutants with different linker lengths (with no or 1-3 PT/S-box units) were successfully constructed and purified. Results showed that the activity of cellulases on Avicel and regenerated amorphous cellulose (RAC) increased with the number of PT/S-box units. Furthermore, the desorption of AcCel12B and its mutants from RAC and Avicel were significantly different. The energy of desorption of wild-type and mutant AcCel12B from cellulose decreased with the number of PT/S-box units. Thus, AcCel12B containing more PT/S-box units was more easily desorbed and had more opportunity to hydrolyze cellulose than other samples. The number of PT/S-box units in endocellulase affected the desorption of the enzyme, which is possibly responsible for the differences in the activity of wild-type and mutant AcCel12B on Avicel and RAC. [ABSTRACT FROM AUTHOR]

Several autoimmune diseases are marked by a deficiency of soluble tumor necrosis factor (TNF). The TNF deficiency is caused in at least one autoimmune disease, multiple sclerosis, by an overabundance of TNF receptor 1 (TNFR1). Excess TNFR1 binds and inactivates TNF and this leaves less TNF bioavailable. This study sought to determine if expression of fresh or IL2-stimulated TNF receptors on Tregs cells, an important immunoregulatory cell involved in autoimmunity, is altered in type I diabetes. Standard fluorescence analysis was used to examine the levels of TNFR1 and TNFR2 on human Tregs in patients with type I diabetes (T1D) or controls. Fresh Tregs from T1D compared to control Tregs had identical levels of TNFR1. In marked contrast, Type 1 diabetic patients Treg cells had statistically elevated levels of TNFR2 compared to controls. Tregs stimulated with IL2 from both T1D and controls showed marked increase of TNFR2 expression in a dose-response manner, but the dose response increase in TNFR2 was significantly higher for T1D Treg cells. No IL2 dose-response was present for TNFR1 on either T1D or control Tregs exposed to IL2. A large study of serum for secreted levels of TNFR2 also revealed elevated circulating levels consistent with the elevated surface expression on Tregs. These findings suggest that abnormal regulation of TNFR2 expression with elevated cellular and secreted levels of TNFR2 is a characteristic of Type 1 diabetes. It is possible that the relative deficiency of TNF in type I diabetes, in contrast to multiple sclerosis, is caused by excess expression of TNFR such as TNFR2, a binding structure for inactivating TNF. [ABSTRACT FROM AUTHOR]

In this study, by comparing the four groups of sausages, namely, CO (without starter culture), LB (with Lactobacillus sakei), LS (with L. sakei 3X-2B + Staphylococcus xylosus SZ-8), and LSS (with L. sakei 3X-2B + S. xylosus SZ-8 + S. carnosus SZ-2), the effects of mixed starter cultures on physical–chemical quality, proteolysis, and biogenic amines (BAs) during fermentation and ripening were investigated. Inoculation of the mixed starter cultures increased the number of lactic acid bacteria and staphylococci in sausages during fermentation and ripening for 0 to 5 days. The L. sakei 3X-2B + S. xylosus SZ-8 + S. carnosus SZ-2 mixed starter accelerated the rate of acid production and water activity reduction of sausages and improved the redness value. Compared with CO, the mixed starter effectively inhibited Enterobacteriaceae. At the end of ripening, the LSS group was approximately 1.25 CFU/g, which was less than the CO group, thereby reducing the total volatile basic nitrogen (TVB-N) in the LSS group. The free amino acids in the LS and LSS groups (224.97 and 235.53 mg/kg dry sausage, respectively) were significantly (p < 0.001) higher than that in the CO group (170.93 mg/kg dry sausage). The level of histamine, cadaverine, putrescine, and common BAs showed an opposite trend to the increase of the corresponding precursor amino acid content, which were significantly lower (p < 0.001) in the LS and LSS sausages than in CO. This study showed that L. sakei 3X-2B + S. xylosus SZ-8 + S. carnosus SZ-2 is a potential mixed starter for fermented meat products. [ABSTRACT FROM AUTHOR]

Simple Summary: Currently, knowledge regarding Apis cerana–Nosema ceranae interaction is very limited, though A. cerana is the original host of N. ceranae. Apis cerana cerana is a subspecies of A. cerana and a major bee species used in the beekeeping industry in China and other countries. Here, the effective infection of A. c. cerana workers by N. ceranae was verified, followed by transcriptomic investigation of host responses. Furthermore, immune responses between A. c. cerana and Apis mellifera ligustica were deeply compared and discussed. In total, 1127 and 957 N. ceranae-responsive genes were identified in the infected midguts at 7 d post-inoculation (dpi) and 10 dpi, respectively. Additionally, DEGs in workers' midguts at both 7 dpi and 10 dpi were associated with six cellular immune pathways and three humoral immune pathways. Noticeably, one up-regulated gene was enriched in the NF-κB signaling pathway in the midgut at 10 dpi. Further analysis indicated that different cellular and humoral immune responses were employed by A. c. cerana and A. m. ligustica workers to combat N. ceranae. Our findings provide a foundation for clarifying the mechanisms regulating the immune response of A. c. cerana workers to N. ceranae invasion and developing new approaches to control bee microsporidiosis. Here, a comparative transcriptome investigation was conducted based on high-quality deep sequencing data from the midguts of Apis cerana cerana workers at 7 d post-inoculation (dpi) and 10 dpi with Nosema ceranae and corresponding un-inoculated midguts. PCR identification and microscopic observation of paraffin sections confirmed the effective infection of A. c. cerana worker by N. ceranae. In total, 1127 and 957 N. ceranae-responsive genes were identified in the infected midguts at 7 dpi and 10 dpi, respectively. RT-qPCR results validated the reliability of our transcriptome data. GO categorization indicated the differentially expressed genes (DEGs) were respectively engaged in 34 and 33 functional terms associated with biological processes, cellular components, and molecular functions. Additionally, KEGG pathway enrichment analysis showed that DEGs at 7 dpi and 10 dpi could be enriched in 231 and 226 pathways, respectively. Moreover, DEGs in workers' midguts at both 7 dpi and 10 dpi were involved in six cellular immune pathways such as autophagy and phagosome and three humoral immune pathways such as the Toll/Imd signaling pathway and Jak-STAT signaling pathway. In addition, one up-regulated gene (XM_017055397.1) was enriched in the NF-κB signaling pathway in the workers' midgut at 10 dpi. Further investigation suggested the majority of these DEGs were engaged in only one immune pathway, while a small number of DEGs were simultaneously involved in two immune pathways. These results together demonstrated that the overall gene expression profile in host midgut was altered by N. ceranae infection and some of the host immune pathways were induced to activation during fungal infection, whereas some others were suppressed via host–pathogen interaction. Our findings offer a basis for clarification of the mechanism underlying the immune response of A. c. cerana workers to N. ceranae infection, but also provide novel insights into eastern honeybee-microsporodian interaction. [ABSTRACT FROM AUTHOR]

5-Methoxyleoligin and leoligin are natural occurring lignans derived from Edelweiss (Leontopodium nivale ssp. alpinum), displaying potent pro-angiogenic and pro-arteriogenic activity. Cholesterol efflux from macrophages is associated with reverse cholesterol transport which inhibits the development of cardiovascular disease. Within this study, we developed a modular and stereoselective total synthesis of 5-methoxyleoligin which can be readily used to prepare a novel compound library of related analogs. The target 5-methoxyleoligin was synthesized exploiting a recently disclosed modular route, which allows also rapid synthesis of analogous compounds. All obtained products were tested towards macrophage cholesterol efflux enhancement and the performance was compared to the parent compound leoligin. It was found that variation on the aryl moiety in 2-position of the furan ring allows optimization of the activity profile, whereas the ester-functionality does not tolerate significant alterations. [ABSTRACT FROM AUTHOR]

Alternatively-activated pathways have been observed in biological experiments in cancer studies, but the concept had not been fully explored in computational cancer system biology. Therefore, an alternatively-activated pathway identification method was proposed and applied to primary breast cancer and breast cancer liver metastasis research using microarray data. Interestingly, the results show that cytokine-cytokine receptor interaction and calcium signaling were significantly enriched under both conditions. TGF beta signaling was found to be the hub in network topology analysis. In total, three types of alternatively-activated pathways were recognized. In the cytokine-cytokine receptor interaction pathway, four active alteration patterns in gene pairs were noticed. Thirteen cytokine-cytokine receptor pairs with inverse activity changes of both genes were verified by the literature. The second type was that some sub-pathways were active under only one condition. For the third type, nodes were significantly active in both conditions, but with different active genes. In the calcium signaling and TGF beta signaling pathways, node E2F5 and E2F4 were significantly active in primary breast cancer and metastasis, respectively. Overall, our study demonstrated the first time using microarray data to identify alternatively-activated pathways in breast cancer liver metastasis. The results showed that the proposed method was valid and effective, which could be helpful for future research for understanding the mechanism of breast cancer metastasis. [ABSTRACT FROM AUTHOR]

Lotus (Nelumbo nucifera Gaertn) is a wetland vegetable famous for its nutritional and medicinal value. Phenolic compounds are secondary metabolites that play important roles in the browning of fresh-cut fruits and vegetables, and chemical constituents are extracted from lotus for medicine due to their high antioxidant activity. Studies have explored in depth the changes in phenolic compounds during browning, while little is known about their synthesis during the formation of lotus rhizome. In this study, transcriptomic analyses of six samples were performed during lotus rhizome formation using a high-throughput tag sequencing technique. About 23 million high-quality reads were generated, and 92.14% of the data was mapped to the reference genome. The samples were divided into two stages, and we identified 23,475 genes in total, 689 of which were involved in the biosynthesis of secondary metabolites. A complex genetic crosstalk-regulated network involved in the biosynthesis of phenolic compounds was found during the development of lotus rhizome, and 25 genes in the phenylpropanoid biosynthesis pathway, 18 genes in the pentose phosphate pathway, and 30 genes in the flavonoid biosynthesis pathway were highly expressed. The expression patterns of key enzymes assigned to the synthesis of phenolic compounds were analyzed. Moreover, several differentially expressed genes required for phenolic compound biosynthesis detected by comparative transcriptomic analysis were verified through qRT-PCR. This work lays a foundation for future studies on the molecular mechanisms of phenolic compound biosynthesis during rhizome formation. [ABSTRACT FROM AUTHOR]

The conversion of organic wastes into biochar via the pyrolysis technique could be used to produce soil amendments useful as a source of plant nutrients. In this study, we investigated the effects of fruit peels and milk tea waste-derived biochars on wheat growth, yield, root traits, soil enzyme activities and nutrient status. Eight amendment treatments were tested: no amendment (CK), chemical fertilizer (CF), banana peel biochar 1% (BB1 + CF), banana peel biochar 2% (BB2 + CF), orange peel biochar 1% (OB1 + CF), orange peel biochar 2% (OB2 + CF), milk tea waste biochar 1% (TB1 + CF) and milk tea waste biochar 2% (TB2 + CF). The results indicated that chlorophyll values, plant height, grain yield, dry weight of shoot and root were significantly (p < 0.05) increased for the TB2 + CF treatment as compared to other treatments. Similarly, higher contents of nutrients in grains, shoots and roots were observed for TB2 + CF: N (61.3, 23.3 and 7.6 g kg−1), P (9.2, 10.4 and 8.3 g kg−1) and K (9.1, 34.8 and 4.4 g kg−1). Compared to CK, the total root length (41.1%), surface area (56.5%), root volume (54.2%) and diameter (78.4%) were the greatest for TB2 + CF, followed by BB2 + CF, OB2 + CF, TB1 + CF, BB1 + CF, OB1 + CF and CF, respectively. However, BB + CF and OB + CF treatments increased β-glucosidase and dehydrogenase, but not urease activity, as compared to the TB + CF amendment, while all enzyme activity decreased with the increased biochar levels. We concluded that the conversion of fruit peels and milk tea waste into biochar products contribute the benefits of environmental and economic issues, and should be tested as soil amendments combined with chemical fertilizers for the improvement of wheat growth and grain yield as well as soil fertility status under field conditions. [ABSTRACT FROM AUTHOR]

A Ni–17Mo–7Cr-based superalloy was laser surface-modified to improve its tribological properties. Si particles were employed as coating materials. Si melted on the surface of the alloy during the process, triggering the formation of Mo6Ni6C carbides and Ni–Si intermetallics. A defect-free coating obtained was mostly made up of primary Mo6Ni6C and γ-Ni31Si12, as well as a eutectic structure of β1-Ni3Si and α-Ni-based solid solution (α-Ni (s.s)). The volume fraction of hard reinforcements (Mo6Ni6C, γ-Ni31Si12, and β1-Ni3Si) reached up to 85% in the coating. High-temperature microstructural stability of the coating was investigated by aging the coating at 1073 K for 240–480 h, to reveal its microstructural evolution. In addition, the mechanical performance of the coating was investigated. The nanoscale elastic modulus and hardness of Mo6Ni6C, γ-Ni31Si12, and α-Ni (s.s) were characterized using the nanoindentation tests. The nanoscratch tests were performed to measure the local wear resistance of the coating. Lastly, the Vickers hardness distribution across the cross-section of the coating before and after thermal exposure was compared. The work performed provides basic information understanding the microstructural evolution and mechanical performance of laser-induced coatings on Ni-based superalloys. [ABSTRACT FROM AUTHOR]

In this work, (Au79Ni17Pd4)96Ti4 (wt.%) filler alloy was designed and employed to join SiC ceramics. The effects of brazing temperature and soaking time on the microstructure and fracture morphology of joints were investigated. The results show that the joint obtained can be described as SiC/reaction layer/braze/reaction layer/SiC. The reaction layer was composed of TiC and Au (Si, Ti). The wettability of the filler alloy toward the SiC ceramics was analyzed. The braze zone was mainly constituted by Pd2Si, Ni2Si, and Au (Ni, Si). A large number of nano-sized TiC particles were distributed within the Au (Ni, Si) layer. The formation mechanism of the braze containing different phases was discussed. The brazing temperature and soaking time had a significant effect on the reaction layer at the SiC/braze interface and TiC particles within the Au (Ni, Si) layer, while they showed a negligible effect on the Pd2Si and Ni2Si within the braze. The inherent reason was also clarified in detail. The joint fractography indicated that a good bonding was achieved between the filler alloy and SiC, while joint fracture was primarily induced by the thermal stresses residing after the brazing cycle. [ABSTRACT FROM AUTHOR]

In this work, (Au 79 Ni 17 Pd₄) 96 Ti₄ (wt.%) filler alloy was designed and employed to join SiC ceramics. The effects of brazing temperature and soaking time on the microstructure and fracture morphology of joints were investigated. The results show that the joint obtained can be described as SiC/reaction layer/braze/reaction layer/SiC. The reaction layer was composed of TiC and Au (Si, Ti). The wettability of the filler alloy toward the SiC ceramics was analyzed. The braze zone was mainly constituted by Pd₂Si, Ni₂Si, and Au (Ni, Si). A large number of nano-sized TiC particles were distributed within the Au (Ni, Si) layer. The formation mechanism of the braze containing different phases was discussed. The brazing temperature and soaking time had a significant effect on the reaction layer at the SiC/braze interface and TiC particles within the Au (Ni, Si) layer, while they showed a negligible effect on the Pd₂Si and Ni₂Si within the braze. The inherent reason was also clarified in detail. The joint fractography indicated that a good bonding was achieved between the filler alloy and SiC, while joint fracture was primarily induced by the thermal stresses residing after the brazing cycle.

Background: l-Phenyllactic acid (l-PLA)-a valuable building block in the pharmaceutical and chemical industry-has recently emerged as an important monomer in the composition of the novel degradable biocompatible material of polyphenyllactic acid. However, both normally chemically synthesized and naturally occurring phenyllactic acid are racemic, and the product yields of reported l-PLA synthesis processes remain unsatisfactory., Methods: We developed a novel recombinant Escherichia coli strain, co-expressing l-lactate dehydrogenase (l-LDH) from Lactobacillus plantarum subsp. plantarum and glucose dehydrogenase (GDH) from Bacillus megaterium , to construct a recombinant oxidation/reduction cycle for whole-cell biotransformation of phenylpyruvic acid (PPA) into chiral l-PLA in an enantioselective and continuous manner., Results: During fed-batch bioconversion with intermittent PPA feeding, l-PLA yield reached 103.8 mM, with an excellent enantiomeric excess of 99.7%. The productivity of l-PLA was as high as 5.2 mM·h -1 per OD 600 (optical density at 600 nm) of whole cells. These results demonstrate the efficient production of l-PLA by the one-pot biotransformation system. Therefore, this stereoselective biocatalytic process might be a promising alternative for l-PLA production., Competing Interests: The authors have declared no conflict of interest.

Silymarin is a hepatoprotective mixture of flavonolignans and flavonoids extracted from the seeds of milk thistle (Silybum marianum L. Gaertn). This study investigates the effect of major bioactive constituents from silymarin, silybin A, silybin B, isosilybin A, isosilybin B, silydianin, silychristin, isosilychristin, and taxifolin, on the expression of ABCA1, an important cholesterol efflux transporter, in THP-1-derived macrophages. Four of the studied compounds, isosilybin A, silybin B, silychristin and isosilychristin, were found to significantly induce ABCA1 protein expression without affecting cell viability. Moreover, isosilybin A, a partial PPARγ agonist, was found to promote cholesterol efflux from THP-1 macrophages in a concentration-dependent manner. These findings first show ABCA1 protein up-regulating activity of active constituents of silymarin and provide new avenues for their further study in the context of cardiovascular disease.