Infectious Disease Diagnosis and Pathogen Identification Platform Based on Multiplex Recombinase Polymerase Amplification-Assisted CRISPR-Cas12a System.

Controlling and mitigating infectious diseases caused by multiple pathogens or pathogens with several subtypes require multiplex nucleic acid detection platforms that can detect several target genes rapidly, specifically, sensitively, and simultaneously. Here, we develop a detection platform, termed Multiplex Assay of RPA and Collateral Effect of Cas12a-based System (MARPLES), based on multiplex nucleic acid amplification and Cas12a ssDNase activation to diagnose these diseases and identify their pathogens. We use the clinical specimens of hand, foot, and mouth disease (HFMD) and influenza A to evaluate the feasibility of MARPLES in diagnosing the disease and identifying the pathogen, respectively, and find that MARPLES can accurately diagnose the HFMD associated with enterovirus 71, coxsackievirus A16 (CVA16), CVA6, or CVA10 and identify the exact types of H1N1 and H3N2 in an hour, showing high sensitivity and specificity and 100% predictive agreement with qRT-PCR. Collectively, our findings demonstrate that MARPLES is a promising multiplex nucleic acid detection platform for disease diagnosis and pathogen identification.

MAIT cells in bacterial infectious diseases: heroes, villains, or both?

Due to the aggravation of bacterial drug resistance and the lag in the development of new antibiotics, it is crucial to develop novel therapeutic regimens for bacterial infectious diseases. Currently, immunotherapy is a promising regimen for the treatment of infectious diseases. Mucosal-associated invariant T (MAIT) cells, a subpopulation of innate-like T cells, are abundant in humans and can mount a rapid immune response to pathogens, thus becoming a potential target of immunotherapy for infectious diseases. At the site of infection, activated MAIT cells perform complex biological functions by secreting a variety of cytokines and cytotoxic substances. Many studies have shown that MAIT cells have immunoprotective effects because they can bridge innate and adaptive immune responses, leading to bacterial clearance, tissue repair, and homeostasis maintenance. MAIT cells also participate in cytokine storm generation, tissue fibrosis, and cancer progression, indicating that they play a role in immunopathology. In this article, we review recent studies of MAIT cells, discuss their dual roles in bacterial infectious diseases and provide some promising MAIT cell-targeting strategies for the treatment of bacterial infectious diseases.

Soy protein nanoparticles prepared by enzymatic cross-linking with enhanced emulsion stability.

Particle-stabilized emulsions have shown increasing potential application in food emulsion systems. Here, soy protein, an abundant and inexpensive plant-based protein, was used to develop nanoparticles for emulsion stabilizer applications. An enzymatic cross-linking method based on microbial transglutaminase (mTG) was developed for the fabrication of soy protein nanoparticles (SPNPs). The emulsion stability was compared between soy protein isolate (SPI) and three different nanoparticles. The size of SPNPs ranged from 10 nm to 40 nm, depending on the production conditions. The emulsions stabilized by SPNPs were stable for at least 20 days at room temperature, whereas the emulsion that was stabilized by SPI showed a significant creaming and phase separation phenomenon. The SPNPs also showed a higher antioxidant and reducing effect compared to SPI. The use of mTG induced cross-linking resulted in the formation of covalent bonding between protein molecules, and led to the formation of nanoparticles with higher stability. The approaches support the utilization of inexpensive and abundant plant-based resources as emulsion stabilizers in food applications.

CEO birth order and corporate social responsibility behaviors: The moderating effect of female sibling and age gap.

Corporate social responsibility (CSR) is one of the most important business strategies which helps enterprises obtain competitive advantage and improve performance. Scholars have conducted many beneficial studies on the driving factors of CSR behaviors from the perspective of CEO traits, but rarely focus on the impact of the CEO's early family experiences. This study aims to fill this research gap by investigating the influence of CEO birth order on firms' CSR behaviors, and further exploring the possible moderating effects of the presence of a female sibling and the age gap between the CEO and the closest sibling. This study takes Chinese non-financial private listed companies from 2010 to 2017 as the research samples, and empirically tests the relationship between CEO birth order and a firm's CSR behaviors. The empirical results show that CEO birth order negatively influences corporate social responsibility behaviors, and this relationship would be weakened when the CEO has a female sibling or the age gap between CEO and the closest sibling is larger. This paper extends the research on personal family factors from the field of social psychology to the business field and finds a new driving factor of corporate social responsibility behavior from the perspective of the CEOs' early family factors.

A novel detection method for the pathogenic Aeromonas hydrophila expressing aerA gene and/or hlyA gene based on dualplex RAA and CRISPR/Cas12a.

Aeromonas hydrophila is an emerging waterborne and foodborne pathogen with pathogenicity to humans and warm water fishes, which severely threatens human health, food safety and aquaculture. A novel method for the rapid, accurate, and sensitive detection of pathogenic A. hydrophila is still needed to reduce the impact on human health and aquaculture. In this work, we developed a rapid, accurate, sensitive, and visual detection method (dRAA-CRISPR/Cas12a), without elaborate instruments, integrating the dualplex recombinase-assisted amplification (dRAA) assay and CRISPR/Cas12a system to detect pathogenic A. hydrophila expressing aerA and/or hlyA virulence genes. The dRAA-CRISPR/Cas12a method has high sensitivity, which can rapidly detect (about 45 min) A. hydrophila with the limit of detection in 2 copies of genomic DNA per reaction, and has high specificity for three pathogenic A. hydrophila strains (aerA+hlyA- , aerA-hlyA+ , and aerA+hlyA+ ). Moreover, dRAA-CRISPR/Cas12a method shows satisfactory practicability in the analysis of the spiked human blood and stool and fish samples. These results demonstrate that our developed pathogenic A. hydrophila detection method, dRAA-CRISPR/Cas12a, is a promising potential method for the early diagnosis of human A. hydrophila infection and on-site detection of A. hydrophila in food and aquaculture.

Characterization of iron reducibility of soy protein amyloid fibrils and their applications in iron fortification.

Iron deficiency is a common nutritional disorder worldwide. Iron fortification of food is an effective strategy to control iron deficiency anemia (IDA), however, traditional iron fortificants usually provoke undesirable organoleptic changes or have limited colloid stability. In this research, we investigated iron reducibility of soy protein amyloid fibrils made from soy protein isolates (SPI), soy ß-conglycinin (7S) and soy glycinin (11S), and explored their applications in iron fortification. All three protein fibrils showed iron reducibility. The reducibility was utilized to generate fibril-iron nanoparticle composites. The iron reducibility was affected by fibril concentration, degree of fibrillation and reducing amino acid composition. We identified 11S had the most significant effect on reducing Fe (III) to more bioavailable Fe (II) state, whereas 7S showed the optimal result for generation of iron nanoparticle on fibrils in situ. The resulted fibril-iron nanoparticle hybrids showed high dispersibility in various liquid foods, without distinct color change.

Enhanced Antibacterial Activity of Hen Egg-White Lysozyme against Staphylococcus aureus and Escherichia coli due to Protein Fibrillation.

Antibacterial agents with broad-spectrum antibacterial properties have always been in large demand. Lysozyme, a common and inexpensive protein, is widely used in food safety and biomedical applications for antibacterial purposes. However, many pathogens are lysozyme-resistant or insensitive. In this research, we investigated the antibacterial activities and mechanism of oligomers and amyloid fibrils formed from hen egg-white lysozyme (HEWL) against Staphylococcus aureus and Escherichia coli. The HEWL fibrils showed significantly enhanced antibacterial activity against both lysozyme-resistant S. aureus and lysozyme-insensitive E. coli. The HEWL oligomers, on the other hand, did not show an obvious improvement in antibacterial activity compared to native HEWL. Our results indicated that the fibrillation of HEWL can significantly enhance antibacterial activity against both S. aureus and E. coli. The natural and inexpensive HEWL amyloid fibrils can be potentially applied to antimicrobial food packaging, animal feed, antibiotic replacement, etc.

Time lag of vegetation growth on the Loess Plateau in response to climate factors: Estimation, distribution, and influence.

Vegetation growth is importantly affected by seasonal weather patterns. The time lag between changes in these patterns and the vegetative response is an important factor in vegetation-climate interactions and can vary considerably with the spatial heterogeneity of an ecosystem. In this study of the Loess Plateau, China, highly accurate time lags were determined at the daily scale using linear regression based on a multiyear normalized difference vegetation index (NDVI) dataset (1982-2015) and the corresponding reconstructed monthly climate series. We found that lag length varied among catchments, vegetation zones, and land-cover types. The mean time lag between precipitation and the NDVI response varied from 7.9 days to 17.7 days across the catchments; for temperature and NDVI, the lag in response ranged from 6.2 days to 25.3 days. Across the six vegetation zones that range from the southeast to the northwest of the Loess Plateau, both the precipitation-NDVI and temperature-NDVI lag lengths were largest in the central zones and lower in the zones closer to the edges of the plateau. Among the different land-cover types, grassland had the largest lags between precipitation or temperature change and the NDVI response, followed by shrubland, arable land, and then forests. The possible impact of vegetative time lags on sediment yield was also investigated for the first time, and this information has great relevance for soil conservation on the Loess Plateau and sediment reduction in the lower Yellow River.

Preparation, characterization, and antibacterial properties of biofilms comprising chitosan and ε-polylysine.

Of late, the demand for food-packaging materials, in particular, multifunction packaging materials that are biodegradable; antibacterial; have good mechanical and barrier properties; and are edible and transparent, has increased considerably. In this study, we prepared chitosan (CS)/ε-polylysine (PL) biofilms with different CS-to-PL ratios. We studied the preparation, mechanical properties, microstructures, thermal stability, transparency, water-vapor permeability, oil permeability, and antibacterial properties of the composite CS/PL biofilms. The results demonstrate that CS/PL biofilms are mechanically strong, have good thermal stability, high transparency, low water-vapor and oil permeability, and extensive antibacterial properties that act against Escherichia coli, Bacillus subtilis, and yeast. Therefore, we therefore conclude that CS/PL biofilms are promising food-packaging materials.

Revisiting the prospects of plastein: thermal and simulated gastric stability in relation to the antioxidative capacity of casein plastein.

Plastein, a product of protease-induced peptide aggregation, is thought to possess unique physical properties and bioactivity, although its formation, stability, and functional mechanisms remain unclear. This study demonstrates that plastein is formed from bovine casein peptides with Alcalase by hydrophobic and electrostatic interactions and less likely by covalent bonding. The peptide aggregation enhanced the Fe(III) reducing potential and decreased the Fe(II) chelating activity (p < 0.05) of casein peptides, but there was no difference in inhibition of Fe-induced linoleic acid peroxidation after plastein reaction. The casein plastein product retained its antioxidative activities after being heated at 100 °C. However, simulated gastric protease treatment with pepsin and pancreatic enzymes resulted in enhanced reducing potential and metal chelation of the casein plastein and reduction of the inhibitory effect on lipid peroxidation. It appears that the plasteins were disintegrated and further hydrolyzed by gastric proteases on the basis of the antioxidative capacity and RP-HPLC profile being similar to those of the casein hydrolysates. Therefore, plastein reaction may not confer metabolic stability or enhance the antioxidative capacity of casein peptides for prospective functional food applications.

[Effects of forest vegetation on runoff and sediment production on sloping lands of Loess area].

Based on the 1985 - 2003 fixed-position data in 9 runoff plots of Caijiachuan watershed in the Jixian county of Shanxi Province in loess area, this paper discussed the relationships between vegetation and runoff and sediment production on sloping lands of loess area. The results showed that natural-secondary forest had better function in soil and water conservation than artifical Robinia pseudoacacia forest, with runoff and sediment produced 65% - 82% and 23% - 92%, respectively. Multiple regression analysis indicated that runoff and sediment production had a significant correlation with rainfall and its intensity, but this relationship was decreased gradually with increasing canopy density. Different land-use type had different runoff and sediment production, e. g., Ostryopsis davidiana and natural-secondary forests had the least runoff and sediment production, artificial Robinia pseudoacacia and Pinus tabulaeformis forests had 5 folds of it as much as Ostryopsis davidiana forest, mixed apple trees and crops had 17.14 and 3.96 folds of it than Ostryopsis davidiana forest, respectively, while high-standard soil preparation could decrease the production obviously. Gray correlation analysis suggested that the stand canopy density and the biomass of herb and litter were the most important factors affecting the runoff and sediment production on sloping land, whose gray correlation degrees all exceeded 0.6. Mixed forest with multi-layer stand structure and shrub forest should be developed in vegetation re-construction of loess area, which could help to increase the coverage and litter thickness to dramatically decrease the runoff and sediment production on sloping land.