Functional identification of the calcineurin B-like protein PavCBL4 in modulating salt tolerance in sweet cherry

Abiotic stresses, such as high salinity, pose a significant threat to plant growth and development, reducing crop yield and quality. Calcineurin B-like (CBL) proteins serve as crucial calcium sensors in plant responses to diverse environmental stresses. However, the CBL family in sweet cherry has not been identified at the genome-wide level, and the regulatory role of CBL proteins in cherry plants’ salt response is unclear. Here, we identified 10 CBL family genes (PavCBLs) from the Prunus avium genome and cloned seven of them. We comprehensively analyzed PavCBL genes for collinearity, phylogenetic relationships, gene structure, and conserved motifs. Expression analysis revealed significant induction of transcription under abiotic stress, with PavCBL4 displaying the most substantial expression change. Additionally, we identified PavCBL4 as a PavSOS2 (Salt Overly Sensitive 2)-interacting protein through Y2H and Split-LUC assays. Subcellular localization analysis indicated that PavCBL4 is present in both the cytoplasm and nucleus. Functional assessment of PavCBL4 in the PavCBL4-overexpressing transgenic ‘Gisela 6’ plants showed its positive role in enhancing salt tolerance in cherry plants. Measurements of Na+ content and antioxidant enzyme activity under salt stress indicated that PavCBL4 functions positively by inhibiting Na+ accumulation and promoting ROS scavenging in response to salt stress. These findings lay the groundwork for a deeper understanding of the molecular mechanisms underlying PavCBL-mediated salt tolerance in sweet cherry

Effects of coated sodium butyrate on the growth performance, serum biochemistry, antioxidant capacity, intestinal morphology, and intestinal microbiota of broiler chickens

IntroductionThis study examined the impact of adding coated sodium butyrate (CSB) to the diet on the growth performance, serum biochemistry, antioxidant capacity, intestinal morphology, and cecal microbiota of yellow-feathered broiler chickens.MethodsIn this study, 240 yellow-feathered broiler chickens at 26 days old were divided into two groups: the control group (CON group) received a standard diet, and the experimental group (CSB group) received a diet with 0.5 g/kg of a supplement called CSB. Each group had 6 replicates, with 20 chickens in each replicate, and the experiment lasted for 36 days.ResultsCompared to the CON group, the CSB group showed a slight but insignificant increase in average daily weight gain during the 26–62 day period, while feed intake significantly decreased. The CSB group exhibited significant increases in serum superoxide dismutase, catalase, and total antioxidant capacity. Additionally, the CSB group had significant increases in total protein and albumin content, as well as a significant decrease in blood ammonia levels. Compared to the CON group, the CSB group had significantly increased small intestine villus height and significantly decreased jejunal crypt depth. The abundance of Bacteroidetes and Bacteroides in the cecal microbiota of the CSB group was significantly higher than that of the CON group, while the abundance of Proteobacteria, Deferribacteres, and Epsilonbacteraeota was significantly lower than that of the CON group.ConclusionThese results suggest that adding CSB to the diet can improve the growth performance and antioxidant capacity of yellow-feathered broiler chickens while maintaining intestinal health

Comparative Study of Different Pretreatment and Combustion Methods on the Grindability of Rice-Husk-Based SiO₂

The rice husk (RH) combustion pretreatment method plays a crucial role in the extraction of nanoscale SiO2 from RH as a silicon source. This study examined the effects of diverse pretreatment methods and combustion temperatures on the particle size distribution of nanoscale high-purity amorphous SiO2 extracted from rice husk ash (RHA) post RH combustion. The experiment was structured using the Taguchi method, employing an L9 (21 × 33) orthogonal mixing table. The median diameter (D50) served as the output response parameter, with the drying method (A), combustion temperature (B), torrefaction temperature (C), and pretreatment method (D) as the input parameters. The results showed the torrefaction temperature (C) as being the predominant factor affecting the D50, which decreased with an increasing torrefaction temperature (C). The optimal parameter combination was identified as A2B2C3D2. The verification test revealed that roasting could improve the abrasiveness of Rh-based silica and reduce the average particle size. Torrefaction at medium temperatures might narrow the size distribution range of RHA-SiO2. We discovered that the purity of silica increased with an increasing roasting temperature by evaluating the concentration of silica in the sample. The production of RHA with silica concentrations up to 92.3% was investigated. X-ray diffraction analysis affirmed that SiO2’s crystal structure remained unaltered across different treatment methods, consistently presenting as amorphous. These results provide a reference for extracting high-value products through RH combustion

Border-rows Effect of Rape (Brassica napus L.) Intercropping with Milk Vetch (Astragalus sinicus L.)

Border-rows effect is an important research content of intercropping system. Milk vetch-rape intercropping is one of the typical intercropping patterns in southern China. However, research on the border-rows effect is very little, which has affected the application and popularization of the milk vetch-rape intercropping system. In this study, two field experiments were conducted from 2018 to 2020. The effects of monoculture, intercropping and different border-rows on agronomic traits and yield of rape were studied. The results showed that milk vetch had a significant effect on the first border-row of rape. The first border-row of rape diameter of root and secondary effective branch number is significantly higher than second and third border-row, and the rape yield per plant in the first border-row of intercropping milk vetch was 135%, 328%, 257% and 147% higher than that in the second, third, fourth and fifth border-rows respectively. The intercropping with milk vetch enhanced the number of pods per plant in first border-row by increasing the agronomic traits of rape, such as plant height, stem diameter, primary effective branch number and secondary effective branch number. In conclusion, intercropping milk vetch significantly improved the agronomic traits of the first and second border-rows rape, increased the yield of rape. So it is recommended that milk vetch intercropping with two rows rape, which has important guiding significance for the application and promotion of milk vetch-rape intercropping in the future

Effects of Dietary <i>Bacillus coagulans</i> and Tributyrin on Growth Performance, Serum Antioxidants, Intestinal Morphology, and Cecal Microbiota of Growing Yellow-Feathered Broilers

This study investigated the impact of Bacillus coagulans (BC) and tributyrin (TB) supplementation on the growth performance, serum antioxidant capacity, intestinal morphology, and cecal microbiota of yellow-feathered broilers. Using a 2 × 2 factorial design, 480 broilers were randomly assigned to four experimental diets, comprising two levels of BC (0 and 1 g/kg) and two levels of TB (0 and 1 g/kg), over a 36-day period. A significant interaction was observed between BC and TB, impacting the average daily feed intake (ADFI) of broilers aged between 26 and 40 days (p p p < 0.05). The addition of BC and TB significantly enhanced the richness and diversity of cecal microbiota, with a notable interactive effect observed for the abundance of Faecalibacterium, Ruminococcus_torques_group, and Phascolarctobacterium. In conclusion, supplementation with BC and TB can effectively improve the growth performance, serum antioxidant capacity, intestinal morphology, and cecal microbiota composition of yellow-feathered broilers, indicating the presence of an interactive effect

Competitive adsorption and desorption of three antibiotics in distinct soil aggregate size fractions

Multiple antibiotics that are used in veterinary medicine coexist in soils, but their interaction and the effects on adsorption and desorption in soils have not been extensively studied. In this study, using batch experiments, we evaluated the adsorption and desorption of sulfadiazine (SDZ), tetracycline (TC), and norfloxacin (NFX) using four different soil aggregate size fractions and discovered that: (1) TC had the highest adsorption (76–98 %) and the lowest desorption in each tested system, whereas SDZ showed opposite adsorption and desorption ability, (2) the highest adsorption and the lowest desorption of all three tested antibiotics were observed with soil macroaggregates (250–2000 µm) in all the cases; in contrast, opposite adsorption and desorption ability were observed for soil clay ( binary system (56–84 %) > ternary system (50–78 %); however, desorption were in the reverse order. The Freundlich equation fitting and Brunauer–Emmett–Teller (BET) analysis further demonstrated that the adsorption competition between the tested antibiotics depended mainly on the specific surface area of each soil aggregate size fractions and its chemical properties. In conclusion, soil macroaggregates play a key role in the retention of antibiotics in soils, and the coexistence of multiple antibiotics greatly increases leaching risk

Comparison of the Metabolic Behaviors of Six Systemic Insecticides in a Newly Established Cell Suspension Culture Derived from Tea (<i>Camellia sinensis</i> L.) Leaves

The use of an <i>in vitro</i> cell suspension to study insecticide metabolism is a simpler strategy compared to using intact plants, especially for a difficult matrix such as tea. In this study, a sterile tea leaf callus was inoculated into B₅ liquid media with 2,4-dichlorophenoxyacetic acid (2,4-D, 1.0 mg L^–1) and Kinetin (KT, 0.1 mg L^–1). After 3–4 subcultures (28 days each), a good cell suspension was established. Utilizing these cultures, the metabolic behaviors of six insecticides, including two organophosphates (dimethoate, omethoate) and four neonicotinoids (thiamethoxam, imidacloprid, acetamiprid, and imidaclothiz) were compared. The results showed that thiamethoxam, dimethoate, and omethoate were easily metabolized by tea cells, with degradation ratios after 75 days of 55.3%, 90.4%, and 100%, respectively. Seven metabolites of thiamethoxan and two metabolites of dimethoate were found in treated cell cultures using mass-spectrometry, compared to only two metabolites for thiamethoxam and one for dimethoate in treated intact plants

Bovine extracellular vesicles contaminate human extracellular vesicles produced in cell culture conditioned medium when ‘exosome-depleted serum’ is utilised

Extracellular vesicles (EVs) are important intercellular communication messengers. Half of the published studies in the field are in vitro cell culture based in which bovine serum in various concentrations and forms is used to facilitate the production of extracellular vesicles. ‘Exosome depleted serum’ is the type of bovine serum most widely used in the production of human EVs. Herein, we demonstrate that, despite the initial caution raised in 2014 about the persistence of bovine EVs, ‘exosome depleted serum’ was still used in 46% of publications on human or rodent EVs between 2015 and 2019. Using nanoparticle tracking analysis combined with detergent lysis of vesicles as well as bovine CD9 ELISA, we show that there were approximately 5.33 x 107/mL of bovine EVs remaining in the ‘exosome depleted serum’. Importantly, the ‘exosome depleted serum’ was relatively enriched in small EVs by approximately 2.7-fold relative to the large EVs compared to that in the original serum. Specifically, the percentage of small EVs in total vesicles had increased from the original 48% in the serum before ultracentrifugation to 92% in the ‘exosome depleted serum’. Furthermore, the pervasive bovine EVs carried over by the ‘exosome depleted serum’, even when the lowest concentration (0.5%) was used in cell culture, resulted in a significant contamination of human EVs in cell culture conditioned medium. Our findings indicate that the use ‘exosome depleted serum’ in cell culture-based studies may introduce artefacts into research examining the function of human and rodent EVs, in particular those involving EV miRNA. Thus, we appeal to the researchers in the EV field to seriously reconsider the practice of using ‘exosome depleted serum’ in the production of human and other mammalian EVs in vitro.</p

Aptamer-guided extracellular vesicle theranostics in oncology

In the past decade, the study of exosomes, nanosized vesicles (50-150 nm) released into the extracellular space via the fusion of multivesicular bodies with the plasma membrane, has burgeoned with impressive achievements in theranostics applications. These nanosized vesicles have emerged as key players in homeostasis and in the pathogenesis of diseases owing to the variety of the cargos they can carry, the nature of the molecules packaged inside the vesicles, and the robust interactions between exosomes and target cells or tissues. Accordingly, the development of exosome-based liquid biopsy techniques for early disease detection and for monitoring disease progression marks a new era of precision medicine in the 21st century. Moreover, exosomes possess intrinsic properties - a nanosized structure and unique &quot;homing effects&quot; - that make them outstanding drug delivery vehicles. In addition, targeted exosome-based drug delivery systems can be further optimized using active targeting ligands such as nucleic acid aptamers. Indeed, the aptamers themselves can function as therapeutic and/or diagnostic tools based on their attributes of unique target-binding and non-immunogenicity. This review aims to provide readers with a current picture of the research on exosomes and aptamers and their applications in cancer theranostics, highlighting recent advances in their transition from the bench to the clinic