In order to evaluate the effects of continuous cropping of millet on soil nutrients and soil enzyme activities, the present study was based on four treatments of 2 years of continuous cropping (T1), 3 years of continuous cropping (T2), 4 years of continuous cropping (T3) and rotational cropping (CK), based on 4 years of no fertilizer positioning experiments, and the soil nutrients, soil enzyme activities and millets yields were determined, respectively. The results showed that with the increase of continuous cropping years, the millet yield decreased and was significantly lower than that of rotating with legume crops, and compared with CK, the yields of T1, T2 and T3 treatments were reduced by 8.92%, 13.73% and 37.60%, respectively; the soil nitrogen and phosphorus contents were reduced, the quick-acting potassium content did not change obviously, and the soil pH was increased; Soil urease, alkaline phosphatase, sucrase and catalase activities generally showed a decreasing trend and the decrease was more significant with the increase in the number of years of continuous cropping. Therefore, in order to maintain the soil fertility and increase the millet yield, it is necessary to practice crop rotation and stubble reversal between millets and leguminous crops such as kidney beans, and to apply certain fertilizers.

Nitraria roborowskii Kom (NRK), with high economic and ecological value, is mainly distributed in the Qaidam Basin, China. However, research on its chemical components and bioactivities is still rare. In this study, its chemical constituents (52) including 10 β-carboline alkaloids, nine cyclic peptides, three indole alkaloids, five pyrrole alkaloids, eight phenolic acids and 17 flavonoids were identified tentatively using UPLC-triple-TOF-MS/MS. Notablely, one new β-carboline alkaloid and five new cyclic peptides were confirmed using MS/MS fragmentation pathways. In addition, experiments in vitro indicated that NRK-C had strong maltase and sucrase inhibitory activities (IC50 of 0.202 and 0.103 mg/mL, respectively). Polysaccharide tolerance experiments confirmed NRK-C (400 mg/kg) was associated with decreased postprandial blood glucose (PBG) in diabetic mice. These results suggested that NRK fruit might be used as a functional ingredient in food products.

Phenolics of mulberry (Morus alba L.) leaves (MLs) have potential anti-diabetic effects, but they may be chemically modified during gastrointestinal digestion so affect their biological activity. In this study, an in vitro digestion model coupled with Caco-2 monolayer and Caco-2/insulin-resistant HepG2 coculture model were used to study the transport and hypoglycemic effects of phenolics in raw MLs (U-MLs) and solid-fermented MLs (F-MLs). The results of LC-MS/MS analysis showed that the Papp (apparent permeability coefficient, 10-6cm/s) of phenolics in digested MLs ranged from 0.002 ± 0.00 (quercetin 3-O-glucoside) to 60.19 ± 0.67 (ferulic acid), indicating higher phenolic acids absorbability and poor flavonoids absorbability. The Papp values of phenolic extracts of F-MLs in Caco-2 monolayer were significantly higher (p > 0.05) than that of U-MLs. Digested phenolic extracts inhibited the activities of sucrase (60.13 ± 2.03 %) and maltase (82.35 ± 0.78 %) and decreased 9.28 ± 0.84 % of glucose uptake in Caco-2 monolayer. Furthermore, a decrease in the mRNA expression of glucose transporters SGLT1 (0.64 ± 0.18), GLUT2 (0.14 ± 0.02) and the sucrase-isomaltase (0.59 ± 0.00) was observed. In Caco-2/insulin-resistant HepG2 co-culture model, phenolic extracts regulated glucose metabolism by up-regulating the mRNA expressions of IRS1 (9.32-fold), Akt (17.07-fold) and GYS2 (1.5-fold), and down-regulating the GSK-3β (0.22-fold), PEPCK (0.49-fold) and FOXO1 (0.10-fold) mRNA levels. Both U-MLs and F-MLs could improve glucose metabolism, and the partial least squares (PLS) analysis showed that luteoforol and p-coumaric acid were the primary phenolics that strongly correlated with the hypoglycemic ability of MLs. Results suggested that phenolics of MLs can be used as dietary supplements to regulate glucose metabolism.

Nitrogen (N), phosphorus (P), and potassium (K) exert various effects on strawberry (Fragaria ananassa Duchesne) yields. In this study, we employed an orthogonal experimental design (T1-T9) with three fertilization treatments (N, P, and K) at three levels to identify an optimal fertilization scheme for strawberry cultivation. The effects of fertilizer combinations the rhizosphere soil microbial community were also explored by using bacterial full-length 16S rRNA and fungal ITS (internal transcribed spacer) sequencing (30 samples for each analysis). The results showed that the average plant height and leaf area of the fertilized groups were 24.6% and 41.6% higher than those of the non-fertilized group (T0). After 60 d of planting, the sucrase activity in the T6 group increased by 76.67% compared to the T0 group, with phosphate fertilizer exerting a more significant impact on sucrase activity. The T6 treatment group had the highest alpha diversity index among bacterial and fungal microorganisms, and had a different microbial community structure compared with the control group. The most abundant bacterial taxa in the strawberry rhizosphere soil were Proteobacteria, Bacteroidota, and Acidobacteriota, and the most abundant fungal phyla were Monoblepharomycota, Glomeromycota, and Mucoromycota. Application of the optimal combined fertilizer treatment (T6) significantly increased the abundance of Proteobacteria and altered the abundance of Gemmatimonas compared to other treatment groups. Notably, Gemmatimonas abundance positively correlated with strawberry plant height and soil N, P, and K levels. These findings indicated that the relative abundance of beneficial bacteria could be enhanced by the application of an optimal fertilizer ratio, ultimately improving strawberry agronomic traits.

Staphylococcus aureus integrated with mecA gene, which codes for penicillin-binding protein 2a, is resistant to all penicillins and other beta-lactam antibiotics, resulting in poor treatment expectations in skin and soft tissue infections. The development of a simple, sensitive and portable biosensor for mecA gene analysis in S. aureus is urgently needed. Herein, we propose a dual-toehold-probe (sensing probe)-mediated exonuclease-III (Exo-III)-assisted signal recycling for portable detection of the mecA gene in S. aureus. When the target mecA gene is present, it hybridizes with the sensing probe, initiating Exo III-assisted dual signal recycles, which in turn release numerous \"3\" sequences. The released \"3\" sequences initiate catalytic hairpin amplification, resulting in the fixation of a sucrase-labeled H2 probe on the surface of magnetic beads (MBs). After magnet-based enrichment of an MB-H1-H2-sucrase complex and removal of a liquid supernatant containing free sucrase, the complex is then used to catalyze sucrose to glucose, which can be quantitatively detected by a personal glucose meter. With a limit of detection of 4.36 fM for mecA gene, the developed strategy exhibits high sensitivity. In addition, good selectivity and anti-interference capability were also attained with this method, making it promising for antibiotic tolerance analysis at the point-of-care.

BACKGROUND: α-Glucosidase inhibitors could effectively reduce postprandial blood glucose (PBG) levels and control the occurrence of complications of diabetes. Gallotannins (GTs) in plants have attracted much attention due to their significant α-glucosidase inhibitory activities in vitro. However, there is still a lack of systematic comparative studies to further elucidate inhibitory activities in vivo and in vitro of these compounds against α-glucosidase, especially for mammalian sucrase and maltase, and analyze their structure-activity relationship.
OBJECTIVE: Determine the in vitro and in vivo inhibitory activities of five GTs with different number of galloyl moieties (GMs) on sucrase, maltase and α-amylase, and elucidate the relationship between α-glucosidase inhibitory activities and the number and connection mode of GMs.
METHODS: Molecular docking and dynamics were used to study the binding mode and binding ability of five GTs against sucrase, maltase and α-amylase. Then, the inhibitory activities and inhibitory mechanisms of these compounds on sucrase, maltase and α-amylase in vitro were studied using inhibitory assay and enzyme inhibition kinetics. Further, the hypoglycemic effects in vivo of these compounds were demonstrated by three polysaccharides tolerance experiments on diabetes model mice.
RESULTS: The results of molecular docking showed that these compounds could bind to enzymes through hydrogen bonds, hydrophobic interactions, etc. In addition, the α-glucosidase inhibition comparative studies in vitro and in vivo demonstrated that the inhibitory activities of these compounds on all three sucrase, maltase and α-amylase were ranked as TA ≈ PGG > TeGG > TGG > 1GG, and their inhibitory activities increases with the increase in the number of GMs. Moreover, the hypoglycemic effects of 1,2,3,4,6-pentagalloylglucose (PGG) and tannic acid (TA) in vitro and in vivo were also confirmed to be equivalent to or even stronger than that of acarbose.
CONCLUSIONS: α-Glucosidase inhibitory activities in vitro and in vivo of GTs were positively correlated with the number of GTs, and the more the number, the stronger the activity. However, PGG with five GTs and TA with ten GTs showed almost identical α-glucosidase inhibitory activities, possibly due to the reduced binding force with the enzyme caused by spatial hindrance.

CRISPR-Cas12a is a powerful and programmable tool that has revolutionized the field of biosensing. However, the construction of a CRISPR-Cas12a-mediated portable system for on-site and quantitative detection of mercury ion (Hg2+) has yet to be explored. By integrating a target-triggered cascade toehold-mediated strand displacement reaction (TSDR) and CRISPR-Cas12a, we herein construct a portable on-site biosensor for the quantitative, sensitive, and selective detection of Hg2+ with a glucose meter. The Hg2+ initiates two cascade TSDRs through the T-Hg2+-T interaction to produce multiple double-stranded DNAs that can activate Cas12a\'s trans-cleavage activity. The Cas12a cleaves the sucrase-modified DNA on the electrode, resulting in the liberation of sucrase into the solution. The freed sucrase can catalyze sucrose to generate glucose, which can be quantitatively monitored by a glucometer. The developed portable biosensor provides a dynamic range of 5 orders of magnitude with a detection limit of 40 fM. This biosensor also displays excellent selectivity and stability for detecting Hg2+. Moreover, environmental water samples are utilized to further verify the robustness and effectiveness of the developed biosensor, highlighting its potential application in environmental monitoring and food safety analysis.

The combined effects of biochar and Chlorella under aquaculture wastewater irrigation in improving saline-alkali soil physicochemical properties, microbial communities, and rice yield, is not yet clear. This study utilized soil physicochemical indicators and gene sequencing to examine the effect of salinity stress, biochar and Chlorella under aquaculture wastewater irrigation on soil properties, bacterial community compositions, and rice production. Treatments included three factors in a randomized complete block design with three replications: (i) Biochar - 40 tons ha -1 (BW) versus no-biochar (BN); (ii) Salinity - 3‰ salinity (SH) versus 1‰ salinity (SL); and (iii) Chlorella - with 107 cells mL -1 Chlorella (CW) versus no-Chlorella (CN). The results revealed that increased salinity adversely affected the soil nutrients (TOC, NO3⁻-N, NH4+-N, Olsen-P), and enzyme activity (urease, sucrase, catalase), resulting in a 9.67% reduction in rice yield compared to SL treatment. However, the close correlation between alterations in soil bacterial communities, functions, and soil physicochemical properties, as well as rice yield, indicated that biochar and Chlorella promoted rice yield by enhancing the physicochemical properties of saline-alkali soil and bacterial community when irrigated with aquaculture wastewater: (1) addition of biochar increased the146.05% rice yield by increasing TOC content, the complexity of bacterial co-occurrence patterns, nitrogen fixation potential, and nitrification potential, (2) addition of Chlorella increased TOC, NO3⁻-N, NH4+-N, enhanced urease, sucrase, catalase activity, and nitrification potential to increased rice yield by 60.29%, and (3) compared with the treatment T3 (SHBNCN), the treatments with biochar (BW) and Chlorella (CW) increased the yield by 561.30% and 445.03% under 1‰ and 3‰ salinity, respectively. These findings provide novel perspectives on the capacity of biochar and Chlorella to improve saline-alkali soil properties and increase rice yield irrigated with aquaculture wastewater.

BACKGROUND: This study examined the changes in soil fertility in a maize cropping area when chemical fertilizer was partially replaced with straw or livestock manure over a 33-year period. Four treatments were included: (i) CK (no fertilizer application); (ii) NPK (only chemical fertilizer application); (iii) NPKM (chemical fertilizer partly replaced with livestock manure); (iv) NPKS (chemical fertilizer partly replaced with straw).
RESULTS: Soil organic carbon increased by 41.7% and 95.5% in the NPKS and NPKM treatments, respectively, over the 33-year trial compared with the initial concentration. However, soil organic carbon in NPK was significantly reduced by 9.8%. Soil total N, P and K increased in both NPKM and NPKS treatments compared to the original soil. Soil pH was significantly acidified from 7.6 to 5.97 in the NPK treatment during the experimental period. The NPKM and NPKS treatments buffered the acidification compared to NPK. Meta-analysis results showed that, compared with NPK, NPKM significantly raised soil bacteria and fungi populations by 38.7% and 58.6%; enhanced microbial biomass carbon and nitrogen by 66.3% and 63%, respectively; and increased sucrase, urease and catalase activities by 34.2%, 48.2% and 21.5%. NPKS significantly increased soil fungi and actinomycetes populations by 24.3% and 41.2%, respectively; enhanced microbial biomass carbon and nitrogen by 27.1% and 45%; and strengthened sucrase and urease activities by 36% and 20.3%, respectively.
CONCLUSIONS: Long-term chemical fertilizer application led to the deterioration of soil fertility and environment. Partial replacement of chemical fertilizers with organic materials could significantly amend and buffer such negative effects. © 2023 Society of Chemical Industry.

Isomaltodextrins (IMDs) are starch-based dietary fibers (DF) prepared enzymatically, which show great potential as a functional food ingredient. In this study, a series of novel IMDs with diverse structures were generated by 4,6-α-glucanotransferase GtfBΔN from Limosilactobacillus fermentum NCC 3057, combined with two α-1,2 and α-1,3 branching sucrases. Results indicated that α-1,2 and α-1,3 branching significantly improved the DF contents of α-1,6 linear products up to 60.9-62.8%. When altering the ratios of [sucrose]/[maltodextrin], IMDs containing 25.8-89.0% α-1,6 bonds, 0-59.6% α-1,2 bonds and 0-35.1% α-1,3 bonds and Mw ranged from 1967 to 4876 Da were obtained. Physicochemical property analysis showed that grafting with α-1,2 or α-1,3 single glycosyl branches can improve the solubility of the α-1,6 linear product, in which α-1,3 branched products were better. Moreover, α-1,2 or α-1,3 branching did no effect on the viscosity of the products but Mw did, the larger Mw the greater viscosity. In addition, α-1,6 linear and α-1,2 or α-1,3 branched IMDs all exhibited strong acid-heating stabilities, freeze-thaw stabilities, and good resistance to browning caused by the Maillard reaction. Branched IMDs showed excellent storage stabilities at room temperature for one year at a concentration of 60%, whereas 45% α-1,6 linear IMD precipitated quickly within 12 h. Most importantly, α-1,2 or α-1,3 branching remarkably increased the contents of resistant starch in the α-1,6 linear IMDs to 74.5-76.8%. These clear qualitative assessments demonstrated the outstanding processing and application properties of the branched IMDs and were expected to provide valuable perspectives toward the technological innovation of functional carbohydrates.