Proteomic Analysis of Milk Fat Globule Membrane Protein Modulation of Differently Expressed Proteins in Lactobacillus plantarum under Bile Salt Stress.

Tolerance to bile stress is a crucial property for lactic acid bacteria (LAB) to survive in the gastrointestinal tract and exert their beneficial effects. Whey powder enriched with milk fat globule membrane proteins (M-WPI) as a functional component is protective for strains under stress conditions. The current study investigated the key mechanisms of action involved in Lactobacillus plantarum (L. plantarum) CGMCC 23701 survival in the presence of bile and the protective mechanism of M-WPI. According to proteomic analysis (proteomics), there could be several reasons for the greater protective effect of M-WPI. These include promoting the synthesis of fatty acids and peptidoglycans to repair the structure of the cell surface, regulating the metabolism of carbohydrates and amino acids to release energy and produce a range of precursors, enabling the expression of the repair system to repair damaged DNA, and promoting the expression of proteins associated with the multidrug efflux pump, which facilitates the exocytosis of intracellular bile salts. This study helps us to better understand the changes in proteome of L. plantarum CGMCC 23701 under bile salt stress and M-WPI protection, which will provide a new method for the protection and development of functional LAB.

A preliminary report on the exploration of salivary bacterial diversity by the multiplex SNaPshot assay.

Salivary bacterial community composition is associated with the host's internal and environmental factors, which have potential applications in forensic practice. The 16S rRNA gene sequencing is the most commonly used strategy for detecting salivary bacterial diversity; however, its platforms are not compatible with capillary electrophoresis (CE) platforms commonly used for forensic applications. Therefore, we attempted to detect the salivary bacterial diversity using a single nucleotide polymorphism (SNP) assay. Salivary bacterial diversity varies among diverse geographic locations, making it a potential supplementary biomarker for forensic geographic sourcing. To evaluate the performance of the multiplex SNaPshot assay, saliva samples from three geographic locations in China were analyzed using the multiplex SNaPshot assay and 16S rRNA gene sequencing. We screened SNPs from two high-relative-abundance salivary genera (Streptococcus and Veillonella) to construct a multiplex SNaPshot system that can be used on the CE platform. The stability and sensitivity of the multiplex SNaPshot system were also tested. A random forest classification model was used to classify samples from different regions to explore the ability of salivary bacteria to discriminate between geographic sources. Six bacterial SNPs were screened and a multiplex SNaPshot system was constructed. The stability results showed that the typing of salivary stains that were placed indoors for different days was not affected in this study. Two-thirds of mocked salivary stain samples showed more than 90% of typing results obtained for salivary stain samples with an input of 0.1 µl saliva. The results of principal coordinate analysis based on salivary bacterial diversity showed significant differences between samples from the three different geographic locations. The accuracy of the random forest classification was 66.67% based on the multiplex SNaPshot assay and 83.33% based on the 16S rRNA gene sequencing. In conclusion, this is the first attempt to detect salivary bacterial diversity using a multiplex SNaPshot bacterial SNP assay. The geographic difference in human salivary bacterial community composition was significant, as revealed by the multiplex SNaPshot assay; however, its performance in discriminating geographic sources was lower than that of 16S rRNA gene sequencing. This strategy based on bacterial SNP loci may favor the detection of human bacterial diversity in common forensic laboratories but requires further exploration in larger sample sizes and more bacterial SNP loci.

Reconstructing bifurcation diagrams of chaotic circuits with reservoir computing.

Model-free reconstruction of bifurcation diagrams of Chua's circuits using the technique of parameter-aware reservoir computing is investigated. We demonstrate that (1) reservoir computer can be utilized as a noise filter to restore the system dynamics from noisy signals; (2) for a single Chua circuit, a machine trained by the noisy time series measured at several sampling states is capable of reconstructing the whole bifurcation diagram of the circuit with a high precision; and (3) for two coupled chaotic Chua circuits with mismatched parameters, the machine trained by the noisy time series measured at several coupling strengths is able to anticipate the variation of the synchronization degree of the coupled circuits with respect to the coupling strength over a wide range. Our studies verify the capability of the technique of parameter-aware reservoir computing in learning the dynamics of chaotic circuits from noisy signals, signifying the potential application of this technique in reconstructing the bifurcation diagram of real-world chaotic systems.

Effects of the Species and Growth Stage on the Antioxidant and Antifungal Capacities, Polyphenol Contents, and Volatile Profiles of Bamboo Leaves.

Bamboo leaves contain high concentrations of various biologically active compounds, such as polyphenols and volatiles, making them attractive as raw resources for antioxidant additives in the food industry. Here, we investigated the total phenolic content (TPC) and total flavonoid content (TFC) of four bamboo leaf extracts from two species (Phyllostachys edulis and Chimonocalamus delicatus) at two growth stages (first and second years). Antioxidant capacity was determined based on the radical-scavenging capacity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+). We also assessed the antifungal capacity based on mycelial growth inhibition of Colletotrichum musae (C. musae), Botrytis cinerea (B. cinereain), and Alternaria alternata (A. alternata). Pearson's correlation coefficients showed that the TPC was significantly (p < 0.01) negatively correlated with the half-maximal inhibitory concentrations against DPPH and ABTS+, whereas the TFC was positively correlated with C. musae and B. cinereain growth inhibition, which suggest that TPC and TFC might be the major contributors to the antioxidant and antifungal capacities of bamboo leaves, respectively. The volatile organic compounds (VOCs) of bamboo leaves were also analyzed using gas chromatography-ion mobility spectrometry. The VOCs included twenty-four aldehydes, eleven alcohols, four furans, seven esters, fifteen terpenes, three ketones, one pyrazine, and thirty unidentified compounds. Principal component analysis, partial least squares discriminant analysis, and hierarchical cluster analysis were performed to assess the differences in the volatile profiles of the four bamboo leaf samples, from which 23 discriminatory VOCs with variable importance in the projection values > 1 were screened, and part of them were impacted by species or growth stage. These findings provide a theoretical foundation for the use of bamboo leaves.

Sarcomatoid hepatocellular carcinoma mimics hepatic abscess on contrast-enhanced ultrasound.

Sarcomatoid hepatocellular carcinoma (SHCC) is a rare subtype of hepatocellular carcinoma characterized by abdominal pain or persistent fever with an inflammatory reaction. Here, we report a case of SHCC mimicking hepatic abscess described by not only ultrasonography but also computer tomography. SHCC is a rare subtype of hepatocellular carcinoma characterized by epithelial and mesenchymal tumor features with sarcomatoid morphology. Here, we report a case of SHCC described by ultrasonography and computer tomography as well as confirmed by pathological examination.

A high-throughput droplet digital PCR system aiming eight DNA methylation targets for age prediction.

The droplet digital Polymerase Chain Reaction (ddPCR) has garnered recognition for its distinctive attribute of absolute quantification. And it has found practical utility in age prediction through DNA methylation profiles. However, a prevalent limitation in current ddPCR methodologies is the restricted capacity to detect only two targets concurrently in most instruments, leading to high costs, sample wastage, and labor-intensive procedures. To address the limitations, a novel high-throughput ddPCR system allowing for the simultaneous detection of eight targets was developed. Through the implementation of a new 8-plex ddPCR assay, coupled with comprehensive linear regression analyses involving primers and probes ratios, diverse inputs of single CpG sites with distinct primers and probes, and varying plex assay configurations, stable DNA methylation values for four CpGs and stable measurement precisions for distinct multiplex systems were consistently observed. These findings pave the way for advancing the field of chemistry science by enabling more efficient and cost-effective methods. Furthermore, the comparative validation of ddPCR and SNaPshot demonstrated a remarkable concordance in results, and the system also displayed well in the field of various aspects, including species specificity, DNA input, and aged samples. In this study, the recommended input of bisulfite-converted DNA was determined to be 10-50 ng due to the double-positive droplets. Notably, the Pearson correlation coefficient squared values of four CpGs were 0.4878 (ASPA), 0.4832 (IGSF1), 0.6881 (COL1A1), and 0.6475 (MEIS1-AS3). And the testing set exhibited a mean absolute error of 4.5923 years, indicating the robustness and accuracy of the age-predictive model.

Explore variation of salivary bacteria across time and geolocations.

Saliva is an informative body fluid that can be found at various crime scenes, and the salivary bacterial community has been revealed it is a potential auxiliary target for forensic identification. However, the variation of salivary bacterial community composition across time and geolocation needs to be explored. The study was designed to be carried out during the winter vacation that was across about 50 days and eight geographic locations. The high throughput sequencing was performed with the V3-V4 region of the16S rRNA gene to explore salivary bacterial community composition. An overall slight fluctuation of the salivary bacteria was observed, which primarily occurred in the relative abundance of the salivary bacterial taxa. The results of principal coordinate analysis and hierarchical clustering showed samples were clustered by the individuals. All individuals could be correctly identified with the random forest model. In summation, although the relative abundance of salivary bacteria varied across the changes of time and geolocation, the individualized characteristic of salivary bacteria remained steady, which is beneficial for the salivary bacterial application in personal identification.

Atomic Insights into the S Poisoning Effects of Single-Atom Catalysts in Li-S Batteries.

Here, we paved a new way to evaluate the susceptibility of M-N4-based single-atom catalysts (SACs) to sulfur poisoning in Li-S batteries. The strong binding strength of M-S in Ti-N4- and V-N4-based SACs is attributed to the high bond order, pronounced d-p hybridization, and differential charge density. However, overly strong binding strength of M-S can create a high energy barrier that prevents the detachment of covered S atoms and induces sulfur poisoning. This can significantly impede the exposure of catalysts to residual reactants during subsequent charge-discharge cycles. Moreover, the sulfur poisoning will dramatically depress the overall catalytic performance of the SACs during the subsequent charge-discharge process, indicating that some compromise should be made between the high catalytic performance and the sulfur poisoning in designing SACs for Li-S batteries.

A forensic population database of autosomal STR and X-STR markers in the Qiang ethnic minority of China.

The Qiang ethnic group is one of the oldest ethnic groups in China and is the most active ethnic group among all the populations along the Tibetan-Yi corridor. They have had a profound impact nationally and internationally. The paternal and maternal genetic feature of the Qiang ethnic group has been revealed, leaving the question of the genetic characteristics from autosomes and X chromosome not answered. The aim of this study was to explore the potential of 36 A-STR (Microreader™ 36A ID System) and 19 X-STR (Microreader™ 19X System) for application in the Qiang population and to elucidate their genetic diversity in southwest China. The cumulative probability of exclusion (CPE) for autosomal STRs is 1-1.3814 × 10-15 and the mean paternity exclusion chance (MEC) for X-STRs is 1-1.7323 × 10-6. Forensic parameters suggest that the STRs analyzed here are well-suited for forensic applications. The results of phylogenetic, interpopulation differentiation, and principal coordinates analysis (PCoA) indicate that the Qiang people have extensive connections with ethnic minorities in China, supporting the view that the Qiang people are the oldest group in the entire Sino-Tibetan language family. The Qiang appeared genetically more associated with most ethnic groups in China, especially the Han. The calculation of random matching probability (RMP) was improved by Fst correction of allele frequencies to make RMP more accurate and reasonable. This study can fill in the gaps in the Qiang STR reference database, providing valuable frequency data for forensic applications and evidence for the Qiang's genetic pattern as an important ancestral position in the Sino-Tibetan populations.

Functional of tongue sole (Cynoglossus semilaevis) gamma-interferon-inducible lysosomal thiol reductase with implications in innate immune reponse depend on CXXC active site.

The enzyme gamma-interferon-inducible lysosomal thiol reductase (GILT) plays an important role in promoting the processing and presentation of major histocompatibility complex (MHC) class II-restricted antigens. It is also involved in MHC I-restricted antigens catalyzing disulfide bond reduction in fishes' adaptive immunity. The open reading frame of tongue sole (Cynoglossus semilaevis) GILT (tsGILT) gene is 771 bp long, encoding 257 amino acids, with a calculated molecular weight of 28.465 kDa and isoelectric point (pI) of 5.35. After induction with lipopolysaccharide, the expression of tsGILT mRNA was upregulated in spleen and kidney and recombinant tsGILT protein transferred to late endosomes and lysosomes in HeLa cells. The refolded tsGILT was capable of catalyzing the reduction of the interchain disulfide bonds against an IgG substrate depend on the active site CXXC motif at residues 75-78. The process of immune response to bacteria challenge needs GILT to catalyze the reduction of disulfide bond and unfolding native protein antigens, promoting their hydrolysis by proteases. Whether a single mutation or a double mutation of active site CXXC at residues75-78, the 3D structure of tsGILT protein has undergone major changes and lost its activity of catalyzing the reduction of the interchain disulfide bonds.

Extraction and characterisation of pigment from Yanzhiguo [Prunus napaulensis (Ser.) Steud.].

Yanzhiguo [Prunus napaulensis (Ser.) Steud] belongs to Rosaceae family and is consumed as wild fruit, pulp and juice. However, its potential for extracting natural pigment has not yet been explored. Herein, the components in the fresh Yanzhiguo pulp were preliminarily analyzed by liquid chromatography coupled to mass spectrometry. And, the optimal pre-treatment conditions were established for further extraction of Yanzhiguo pigment based on the a* value. Then, by combining the data from single-factor experiments and response surface methodology, the optimal extraction process was established as: 35% EtOH, a liquid-solid ratio of 200:1 mL g-1, an extraction time of 65 min, and an extraction temperature of 100 °C. Moreover, it was found that the a* value and yield had high fitness except when extracted into ethanol (EtOH) with different concentrations. Meanwhile, our result demonstrated Yanzhiguo pigment had high stability in general environments with carmine (a synthetic pigment) as control, except for extreme environments such as direct (hot) sunlight, high temperature (75 °C) and strong alkaline (pH ≥ 11). Also, Yanzhiguo pigment exhibited good antioxidant activity. Our results contribute to more information on Yanzhiguo pigment and promote its application by providing efficient extraction technology.

The combined effects of ultrasound and plasma-activated water on microbial inactivation and quality attributes of crayfish during refrigerated storage.

In this study, a decontamination technology combining ultrasound (US) and plasma-activated water (PAW) was developed to better preserve crayfish. First, the decontamination efficacy of US, PAW and their combinations (UP) on crayfish was quantified after 0, 20, 40, or 60 min of treatments. The total viable count (TVC) was reduced by 0.27-0.77 Log CFU/g after individual US or PAW treatments, while a TVC reduction of 1.17 Log CFU/g was achieved after 40 min of UP treatment. Besides, the changes in psychrotrophic bacteria, lactic acid bacteria, yeasts and molds followed a similar trend to TVC. UP treatments normally resulted in more significant reductions in the natural microbiota of crayfish than US or PAW treatments. Furthermore, the microbial quality, physicochemical properties and sensory properties of crayfish after different treatments were assessed during storage at 4 °C for 12 days. According to TVC and total volatile basic nitrogen (TVB-N) values, the control group became unacceptable from 4 days, US or PAW groups became unacceptable from 6 days, while UP group extended the storage time to 8-10 days. During storage, thiobarbituric acid reactive substances (TBARS) values of all the groups were maintained below 0.5 mg/kg, among which the control group exhibited the highest value (0.39 mg/kg). Moreover, UP treatment effectively retarded the deterioration in color and texture properties of crayfish. Fourier transform infrared (FTIR) spectroscopy analysis indicated that UP treatment decreased the α-helix contents and increased the ß-sheet contents of crayfish proteins, while the structural changes were not evident at the end of storage. Low-field nuclear magnetic resonance (LF-NMR) analysis revealed that UP treatment reduced the water migration and enhanced the stability of bond water in crayfish. In addition, E-nose analysis revealed the protection of UP treatment on the sensory properties of crayfish during storage. This study demonstrated that the combinations of US and PAW treatments effectively accelerated the decontamination of crayfish and contributed to better storage quality.

Transportation of whey protein-derived peptides using Caco-2 cell model and identification of novel cholesterol-lowering peptides.

Background: The increasing morbidity and mortality of cardiovascular disease have become a major factor in human death. Serum cholesterol is considered to be an important risk factor for inducing coronary heart disease, atherosclerosis and other cardiovascular diseases. To screen intestinal absorbable functional small peptides with cholesterol-lowering activity by enzymatic hydrolysis of whey protein and develop cholesterol-based functional food that may become a substitute for chemically synthesized drugs, providing new ideas for diseases caused by high cholesterol. Objective: This study aimed to evaluate the cholesterol-lowering activity of intestinal absorbable whey protein-derived peptides hydrolyzed by alkaline protease, trypsin and chymotrypsin, respectively. Method: The whey protein hydrolysates acquired by enzymatic hydrolysis under optimal conditions were purified by a hollow fiber ultrafiltration membrane with a molecular weight cutoff of 10 kDa. The fractions obtained by Sephadex G-10 gel filtration chromatography were transported through a Caco-2 cell monolayer. The transported peptides were detected in the basolateral aspect of Caco-2 cell monolayers using ultra- performance liquid chromatography-tandem mass spectrometry (UPLC-MS). Results: His-Thr-Ser-Gly-Tyr (HTSGY), Ala-Val-Phe-Lys (AVFK) and Ala-Leu-Pro-Met (ALPM) were unreported peptides with cholesterol-lowering activity. The cholesterol-lowering activities of the three peptides did not change significantly during simulated gastrointestinal digestion. Conclusion: This study not only provides theoretical support for the development of bioactive peptides that can be directly absorbed by the human body, but also provides new treatment ideas for hypercholesterolemia.

A bibliometric analysis of microbial forensics from 1984 to 2022: progress and research trends.

Microbial forensics is a rapidly evolving discipline that has gained significant momentum in recent years. The study evaluated relevant results over the last four decades from 1984 to 2022 all over the world, aiming to analyze the growing trends and research orientations of microbial forensics. Using "microbial forensics" as the search topic in the Web of Science Core Collection, the systematic retrieval identified 579 documents relevant to the field and draw many statistical tables and maps to make the retrieval results visible. According to further bibliometric analysis, there are an increasing number of publications related to microbial forensics from the overall trend, with the highest number of publications recorded in 2021. In terms of the total number of articles, the USA and China were both the leading contributors to the field among 40 countries. The field has developed rapidly in recent years based on the development of next-generation sequencing. Over the course of its development, there are rich keywords in the research of scholars, which focus on diversity and identification. Moreover, despite the early hot topic being PCR (the use of PCR to probe microorganisms), in recent years, the topics, markers, and the potential application of microorganisms in forensic practice have become hot, which also indicates the future research directions of microbial forensic.

Characterization of loss of chromosome Y in peripheral blood cells in male Han Chinese patients with schizophrenia.

BACKGROUND: Schizophrenia (SCZ) has a global prevalence of 1% and increases the risk of mortality, reducing life expectancy. There is growing evidence that the risk of this disorder is higher in males than in females and it tends to develop in early adulthood. The Y chromosome is thought to be involved in biological processes other than sex determination and spermatogenesis. Studies have shown that loss of chromosome Y (LOY) in peripheral blood cells is associated with a variety of diseases (including cancer) and increased all-cause mortality. An analysis of the relationship between LOY and schizophrenia is warranted. METHODS: A total of 442 Chinese males (271 patients with schizophrenia vs. 171 controls) were included in this study. The copy numbers of the Y and X chromosomes were detected by positive droplets targeting the amelogenin gene (AMEL) on the Y chromosome and X chromosome (AMELY and AMELX, respectively), using droplet digital PCR (ddPCR). The LOY percentage was defined as the difference between the concentration of AMELX and the concentration of AMELY divided by the concentration of AMELX, denoted as (X - Y)/X. RESULTS: In the Han Chinese population, the LOY percentage was higher in the schizophrenia group than in the control group (p < 0.05), although there was no significant difference in the presence of LOY between the two groups. A strong correlation was found between the average of the disease duration and the average of the LOY percentage (R2 = 0.506, p = 0.032). The logistic regression analysis implied that the risk of LOY increases by 0.058 and 0.057 per year according to age at onset and duration of disease, respectively (ponset = 0.013, pduration = 0.017). CONCLUSIONS: In the Han Chinese population, the LOY percentage of the disease group was significantly different from that of the control group. The age of onset and duration of schizophrenia might be risk factors for LOY in peripheral blood cells. A larger sample size and expanded clinical information are needed for more in-depth and specific analyses.

High-Performance, Degradable, Self-Healing Bio-Based Nanocomposite Coatings with Antibacterial and Antioxidant Properties.

The purpose of this study is to obtain a bio-based coating with good functional activity and self-healing ability, demonstrating its potential in food, materials, and other application fields. Plastic coatings can cause serious environmental pollution. It was a good solution to replace plastic coatings with degradable coatings. However, the development of degradable coatings in the fields of food and materials was limited due to their insufficient antibacterial ability and weak comprehensive properties. Therefore, chitosan nanoparticles (NPs) loaded with gallic acid (GA) were self-assembled with gelatin (GE) to prepare high-performance, degradable, self-healing bio-based nanocomposite coatings with antibacterial and antioxidant properties. The oxygen permeability of GE nanocomposite coatings decreased gradually with the addition of NPs, and the barrier properties increased significantly. At the same time, due to the excellent antioxidant and antibacterial ability of GA, the antioxidant effect of the nanocomposite coatings increased by 119%, and the antibacterial rate against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) increased by 32% and 58%, respectively, compared with the pure GE coatings. In addition, the nanocomposite coatings can be repaired within 24 h after being scratched at room temperature. Finally, GA coated with chitosan nanoparticles can significantly delay the escape of GA, and the retardation of gallic acid release exceeded 89% in simulated solutions after 24 h immersion, extending the service life of the nanocomposite coatings.

Effect of postharvest 1-methylcyclopropene application on reactive oxygen species scavenging and sucrose metabolism in Gynura bicolor DC.

After harvest, the metabolism of Gynura bicolor DC (G. bicolor) is vigorous, resulting in sugar scarcity and reactive oxygen species (ROS) accumulation, thus aggravating the quality deterioration. 1-Methylcyclopropene (1-MCP) shows crucial effect in alleviating the postharvest metabolism of vegetables and fruits. This research aimed to evaluate the effect of 1-MCP on ROS scavenging and sucrose metabolism in G. bicolor. In this research, G. bicolor was treated with 10 µL L-1 1-MCP for 12 h, followed by storage at 20 ± 2 °C and 90 ± 5% relative humidity in darkness for 7 days. During storage, the increases in the respiration rate, electrolytic leakage, weight loss rate, ROS levels, and membrane lipid oxidation were effectively inhibited by 1-MCP. Moreover, starch and hexose degradation was decreased in the 1-MCP group, as were sucrose synthesis and catabolism. Correlation analysis indicated that sugar starvation was associated with respiration, activities regulation of CAT, SOD, and enzymes involved in sucrose metabolism were associated with the levels of hydrogen peroxide at the early storage. In conclusion, 1-MCP delayed postharvest quality deterioration of G. bicolor by alleviating respiration, inducing oxidative stress to enhance ROS scavenging, and inhibiting sucrose metabolism.

Developing a male-specific age predictive model based on Y-CpGs for forensic analysis.

In forensic work, predicting the age of the criminal suspect or victim could provide beneficial clues for investigation. Epigenetic age estimation based on age-correlated DNA methylation has been one of the most widely studied methods of age estimation. However, almost all available epigenetic age prediction models are based on autosomal CpGs, which are only applicable to single-source DNA samples. In this study, we screened the available methylation data sets to identify loci with potential to meet the objectives of this study and then established a male-specific age prediction model based on 2 SNaPshot systems that contain 13 Y-CpGs and the mean absolute deviation (MAD) values were 4-6 years. The multiplex methylation SNaPshot systems and age-predictive model have been validated for sensitivity (the DNA input could be as low as 0.5 ng) and male specificity. They are supposed to have feasibility in forensic practice. In addition, it demonstrated that the method was also applicable to bloodstains, which were commonly found at crime scenes. The results showed good performance (the training set: R2 = 0.9341, MAD = 4.65 years; the test set: R2 = 0.8952, MAD = 5.73 years) in case investigation for predicting male age. For mixtures, when the male to female DNA ratio is 1:1, 1:10, the deviation between the actual age and the predicted age obtained by the model was less than 8 years, which offers great hope for future prediction of the age of males in mixtures and will be a powerful tool for special cases, such as sexual assault. Furthermore, the work provides a basis for the application of Y-CpGs in forensic science.