Choline Dehydrogenase Contributes to Salt Tolerance in Dunaliella through Betaine Synthesis.

In Dunaliella tertiolecta, a microalga renowned for its extraordinary tolerance to high salinity levels up to 4.5 M NaCl, the mechanisms underlying its stress response have largely remained a mystery. In a groundbreaking discovery, this study identifies a choline dehydrogenase enzyme, termed DtCHDH, capable of converting choline to betaine aldehyde. Remarkably, this is the first identification of such an enzyme not just in D. tertiolecta but across the entire Chlorophyta. A 3D model of DtCHDH was constructed, and molecular docking with choline was performed, revealing a potential binding site for the substrate. The enzyme was heterologously expressed in E. coli Rosetta (DE3) and subsequently purified, achieving enzyme activity of 672.2 U/mg. To elucidate the role of DtCHDH in the salt tolerance of D. tertiolecta, RNAi was employed to knock down DtCHDH gene expression. The results indicated that the Ri-12 strain exhibited compromised growth under both high and low salt conditions, along with consistent levels of DtCHDH gene expression and betaine content. Additionally, fatty acid analysis indicated that DtCHDH might also be a FAPs enzyme, catalyzing reactions with decarboxylase activity. This study not only illuminates the role of choline metabolism in D. tertiolecta's adaptation to high salinity but also identifies a novel target for enhancing the NaCl tolerance of microalgae in biotechnological applications.

Functional Characterization of a CruP-Like Isomerase in Dunaliella.

Dunaliella bardawil is a marine unicellular green algal that produces large amounts of ß-carotene and is a model organism for studying the carotenoid synthesis pathway. However, there are still many mysteries about the enzymes of the D. bardawil lycopene synthesis pathway that have not been revealed. Here, we have identified a CruP-like lycopene isomerase, named DbLyISO, and successfully cloned its gene from D. bardawil. DbLyISO showed a high homology with CruPs. We constructed a 3D model of DbLyISO and performed molecular docking with lycopene, as well as molecular dynamics testing, to identify the functional characteristics of DbLyISO. Functional activity of DbLyISO was also performed by overexpressing gene in both E. coli and D. bardawil. Results revealed that DbLyISO acted at the C-5 and C-13 positions of lycopene, catalyzing its cis-trans isomerization to produce a more stable trans structure. These results provide new ideas for the development of a carotenoid series from engineered bacteria, algae, and plants.

Giant Deformation Induced Staggered-Layer Structure Promoting the Thermoelectric and Mechanical Performance in n-Type Bi2(Te, Se)3.

Bismuth telluride has long been recognized as the most promising near-room temperature thermoelectric material for commercial application; however, the thermoelectric performance for n-type Bi2(Te, Se)3-based alloys is far lagging behind that of its p-type counterpart. In this work, a giant hot deformation (GD) process is implemented in an optimized Bi2Te2.694Se0.3I0.006+3 wt%K2Bi8Se13 precursor and generates a unique staggered-layer structure. The staggered-layered structure, which is only observed in severely deformed crystals, exhibits a preferential scattering on heat-carrying phonons rather than charge-carrying electrons, thus resulting in an ultralow lattice thermal conductivity while retaining high-weight carrier mobility. Moreover, the staggered-layer structure is located adjacent to the van der Waals gap in Bi2(Te, Se)3 lattice and is able to strengthen the interaction between anion layers across the gap, leading to obviously improved compressive strength and Vickers hardness. Consequently, a high peak figure of merit ZT of ≈ 1.3 at 423 K, and an average ZT of ≈ 1.2 at 300-473 K can be achieved in GD sample. This study demonstrates that the GD process can successfully decouple the electrical and thermal transports with simultaneously enhanced mechanic performance.

Variations of the fungal microbiome in Corydalis Rhizoma with different collection areas, processing methods, and storage conditions.

Corydalis Rhizoma (CR, Yanhusuo in Chinese) has been widely used as an analgesic in herbal medicine and functional food. Cases of fungal and mycotoxin contamination in CR have been reported. In this study, the composition and diversity of fungal microbiome in CR samples from four herbal markets and two processing methods were investigated by DNA metabarcoding. Variations of the fungal microbiome in CR during cold and conventional storage were monitored. Results showed that Aspergillus was the dominant genus and saprotroph was the dominant trophic mode. Six potential toxigenic fungi, namely, Aspergillus fumigatus, Aspergillus ostianus, Aspergillus terreus, Penicillium citrinum, Penicillium oxalicum, and Trichothecium roseum, were detected. Differences in fungal composition and diversity among various groups based on collection areas and processing methods were also observed. Moreover, the relative abundance of dominant genera in CR samples stored at different temperatures was significantly different and changed with storage time. This study is the first to reveal the influence of collection areas, processing methods, and storage conditions on the fungal microbiome in CR, which was expected to provide a basis for control strategies of fungal contamination in the industrial chain of CR.

DNA metabarcoding analysis of fungal community on surface of four root herbs.

Objective: Angelicae Sinensis Radix (ASR, Danggui in Chinese), Cistanches Herba (CH, Roucongrong in Chinese), Ginseng Radix et Rhizoma (PG, Renshen in Chinese), and Panacis Quinquefolii Radix (PQ, Xiyangshen in Chinese), widely used as medicine and dietary supplement around the world, are susceptible to fungal and mycotoxin contamination. In this study, we aim to analyze their fungal community by DNA metabarcoding. Methods: A total of 12 root samples were collected from three main production areas in China. The samples were divided into four groups based on herb species, including ASR, CH, PG, and PQ groups. The fungal community on the surface of four root groups was investigated through DNA metabarcoding via targeting the internal transcribed spacer 2 region (ITS2). Results: All the 12 samples were detected with fungal contamination. Rhizopus (13.04%-74.03%), Aspergillus (1.76%-23.92%), and Fusarium (0.26%-15.27%) were the predominant genera. Ten important fungi were identified at the species level, including two potential toxigenic fungi (Penicillium citrinum and P. oxalicum) and eight human pathogenic fungi (Alternaria infectoria, Candida sake, Hyphopichia burtonii, Malassezia globosa, M. restricta, Rhizopus arrhizus, Rhodotorula mucilaginosa, and Ochroconis tshawytschae). Fungal community in ASR and CH groups was significantly different from other groups, while fungal community in PG and PQ groups was relatively similar. Conclusion: DNA metabarcoding revealed the fungal community in four important root herbs. This study provided an important reference for preventing root herbs against fungal and mycotoxin contamination.

Enantioselective effects of chiral prothioconazole and its metabolites: Oxidative stress in HepG2 cells and lysozyme activity.

Chiral pesticides may exhibit enantioselectivity in terms of bioconcentration, environmental fate, and reproductive toxicity. Here, chiral prothioconazole and its metabolites were selected to thoroughly investigate their enantioselective toxicity and mechanisms at the molecular and cellular levels. Multispectral techniques revealed that the interaction between chiral PTC/PTCD and lysozyme resulted in the formation of a complex, leading to a change in the conformation of lysozyme. Meanwhile, the effect of different conformations of PTC/PTCD on the conformation of lysozyme differed, and its metabolites were able to exert a greater effect on lysozyme compared to prothioconazole. Moreover, the S-configuration of PTCD interacted most strongly with lysozyme. This conclusion was further verified by DFT calculations and molecular docking as well. Furthermore, the oxidative stress indicators within HepG2 cells were also affected by chiral prothioconazole and its metabolites. Specifically, S-PTCD induced more substantial perturbation of the normal oxidative stress processes in HepG2 cells, and the magnitude of the perturbation varied significantly among different configurations (P > 0.05). Overall, chiral prothioconazole and its metabolites exhibit enantioselective effects on lysozyme conformation and oxidative stress processes in HepG2 cells. This work provides a scientific basis for a more comprehensive risk assessment of the environmental behaviors and effects caused by chiral pesticides, as well as for the screening of highly efficient and less biotoxic enantiomeric monomers.

DNA Metabarcoding Reveals the Fungal Community on the Surface of Lonicerae Japonicae Flos, an Edible and Medicinal Herb.

Lonicerae Japonicae Flos (LJF) has been globally applied as an herbal medicine and tea. A number of reports recently revealed fungal and mycotoxin contamination in medicinal herbs. It is essential to analyze the fungal community in LJF to provide an early warning for supervision. In this study, the fungal community in LJF samples was identified through DNA metabarcoding. A total of 18 LJF samples were collected and divided based on the collection areas and processing methods. The results indicated that Ascomycota was the dominant phylum. At the genus level, Rhizopus was the most abundant, followed by Erysiphe and Fusarium. Ten pathogenic fungi were detected among the 41 identified species. Moreover, Rhizopus, Fusarium, and Aspergillus had lower relative abundances in LJF samples under oven drying than under other processing methods. This work is expected to provide comprehensive knowledge of the fungal community in LJF and a theoretical reference for enhanced processing methods in practical manufacturing.

Enantioselective effect of chiral prothioconazole on the conformation of bovine serum albumin.

As a typical chiral triazole fungicide, the enantioselective toxicity of prothioconazole to environmental organisms is of increasing concern. Herein, the binding mechanism of chiral PTCs to BSA was investigated by multi-spectral technique and molecular docking. Fluorescence titration and fluorescence lifetime experiments fully established that quenching BSA fluorescence by chiral PTCs is static quenching and could spontaneously bind to BSA. Hydrophobic interactions dominate the binding process of chiral PTCs to BSA. Differently, although both chiral PTCs and BSA have a primary binding site, the difference in chiral isomerism leads to a stronger binding ability of S-PTC than R-PTC. Both configurations of PTC can change the conformation of BSA and induce changes in the microenvironment around its amino acid residues, and the effect of S-PTC is more significant. Overall, S-PTC exhibited a more substantial effect on BSA structure relative to R-PTC. That is, S-PTC may lead to more potent potential toxicological effects on environmental organisms. This study provides a comprehensive assessment of the environmental behavior of chiral pesticides and their potential toxicity to environmental organisms at the molecular level and provides a theoretical basis for the screening of highly effective and biologically less toxic enantiomers of chiral pesticides.

A reversible CHEF-based NIR fluorescent probe for sensing Hg2+ and its multiple application in environmental media and biological systems.

Hg2+ poses a great threat to human health and the environment due to its bioaccumulation and permanent damage. Herein, a reversible CHEF-based near-infrared fluorescent probe 2-(3-((E)-4-((E)-4-(diethylamino)-2- hydroxybenzylidene)amino)styryl)-5,5-dimethylcyclohex-2-en-1-ylidene)propanedinitrile (DHEY) capable of specifically recognizing Hg2+ was constructed. DHEY exhibits advantages of large Stokes shift (157 nm), excellent selectivity, high sensitivity (LOD = 3.2 µg/L), and fast response efficiency (<3 min). Interestingly, DHEY can also realize rapid and effective detection of Hg2+ after being recycled 7 times. The successful recovery of trace Hg2+ in different environmental water samples fully demonstrates the potential of DHEY for actual applications. In particular, DHEY enables real-time observation of the distribution and translocation pattern of exogenous Hg2+ in HeLa cells and zebrafish. This work provides important theoretical support for investigating the fate of heavy metal ions in the environment using fluorescence techniques.

Potential toxic effects of sulfonamides antibiotics: Molecular modeling, multiple-spectroscopy techniques and density functional theory calculations.

Sulfonamide antibiotics (SAs) are widely used in medicine, animal husbandry and aquaculture, and excessive intake of SAs may pose potential toxicity to organisms. The toxicological mechanisms of two classical SAs, sulfamerazine (SMR) and sulfamethoxazole (SMT), were investigated by molecular docking, DFT and multi-spectroscopic techniques using HSA and BSA as model proteins. The quenching of HSA/BSA endogenous fluorescence by SMR was higher than that by SMT due to the stronger binding effect of the pyrimidine ring on HSA/BSA compared to the oxazole ring, and that result was consistent with that predicted by DFT calculations. Thermodynamic parameters show that the binding of SAs to HSA/BSA is an exothermic process that proceeds spontaneously (ΔG < 0). Marker competition experiments illustrate that the binding site of SMR/SMT on serum albumin is located in subdomain IIIA. The combination of SAs and HSA/BSA is mainly realized by hydrogen bond and hydrophobic interaction, and the concept is also supported by molecular modeling. The reduced α-helix content of HSA/BSA induced by SMR/SMT indicates a greater stretching of the protein α-helix structure of the SMR/SMT-HSA/BSA. The results could provide useful toxicological information on the hazards of SAs in response to growing concern that SAs may pose a toxic threat to organisms.

Characterization of the Fungal Community in Fritillariae Cirrhosae Bulbus through DNA Metabarcoding.

Fritillariae Cirrhosae Bulbus (FCB) is a well-known and precious traditional Chinese medicine with a medicinal history spanning thousands of years. In recent years, it has been reported that fungal and mycotoxin contamination influenced the safety and quality of FCB. It is essential to systematically study the fungal community for the early warning of fungal and mycotoxin contamination in this herb. A total of 15 FCB samples were collected from five provinces in China, and the fungal communities in the FCB samples were analyzed via amplifying the internal transcribed spacer 2 region through the Illumina Miseq PE300 platform. Furthermore, we compared the differences in fungal community in five groups based on collection areas. Results showed that Ascomycota (41.58-99.66%) and Mucoromycota (0-57.42%) were dominant at the phylum level. Eurotiomycetes (8.49-63.93%), Eurotiales (8.49-63.53%), and Aspergillaceae (8.49-63.51%) were the most abundant at the class, order, and family levels. Aspergillus (8.49-63.41%), Rhizopus (0-57.42%), Fusarium (0-22.81%), Cladosporium (0.16-9.14%), and Alternaria (0.06-17.95%) were the main genera in FCB samples. A total of 34 fungal taxa were identified at the species level, including five potentially toxigenic fungi namely Penicillium brevicompactum, P. citrinum, P. oxalicum, Trichothecium roseum, and Aspergillus restrictus. The differences in fungal community between the five groups were observed. Our findings provide references for the safe utilization and quality improvement of FCB.

Analysis of Fungal Microbiomes in Edible Medicinal Morindae Officinalis Radix and Alpiniae Oxyphyllae Fructus Using DNA Metabarcoding.

Morindae Officinalis Radix (MOR) and Alpiniae Oxyphyllae Fructus (AOF) have been widely used as dietary supplements and traditional herbal medicines for centuries. Fungal and mycotoxin contamination in MOR and AOF has been reported recently. In this study, fungi in MOR and AOF are first investigated using DNA metabarcoding, and the differences in fungal microbiome between moldy and non-moldy samples are analyzed. The results show that Ascomycota is the most prevailing fungus at the phylum level in MOR and AOF with relative abundances of 49.53-94.32% and 14.81-81.85%, respectively. Penicillium (1.86-76.14%), Cladosporium (1.82-56.65%), and Trichoderma (0.12-19.71%) are the dominant genera in MOR. Penicillium (0.27-56.06%), Papiliotrema (0.04-51.71%), and Cladosporium (3.08-44.41%) are the dominant genera in AOF. Two potential toxigenic fungi were detected, namely, Trichoderma atroviride and Fusarium equiseti. Moreover, the differences in fungal communities between moldy and non-moldy samples were monitored. In conclusion, DNA metabarcoding can be used to assess the fungal microbiome in edible medicinal herbs, thereby providing a basis for ensuring food safety and drug efficacy.

Investigation of fungal contamination in medicinal and edible Lycii Fructus through DNA metabarcoding.

AIMS: Lycii Fructus (LF) is considered as a 'superfood' due to its health benefits and delicious tastes, which has gained popularity worldwide. However, LF is also a proper host for fungal growth due to its abundant nutrients. Fungal contamination seriously affects the quality and safety of LF and poses threats to consumer health. METHODS AND RESULTS: In this study, a total of 15 LF samples were collected from five provinces in China, and were divided into five groups based on the collection areas. Fungal contamination in LF was investigated by targeting the internal transcribed spacer 2 region using Illumina Miseq PE300 platform, and the differences of fungal community in groups based on collection areas were compared. Results showed that the fungal contamination was detected in all the 15 LF samples. Ascomycota, Dothideomycetes, Pleosporales and Pleosporaceae were dominant at the phylum, class, order and family levels, respectively. At the genus level, Alternaria, Cladosporium and Fusarium were the three dominant genera. In all, 24 fungal species were identified. Among which, two species, namely Penicillium oxalicum and Trichothecium roseum, were potentially toxigenic. CONCLUSIONS: All 15 LF samples were detected with fungal contamination. The differences of fungal community in LF samples collected from different areas were observed. DNA metabarcoding was demonstrated as an efficient method to monitor the fungal contamination in LF. SIGNIFICANCE AND IMPACT OF THE STUDY: This work comprehensively reveals the fungal diversity and composition in LF and provides early warning for potential mycotoxin contamination.

Illumina-Based Analysis Yields New Insights Into the Fungal Contamination Associated With the Processed Products of Crataegi Fructus.

Crataegi Fructus, a medicinal and edible herb in China, has been considered a popular dietary supplement globally. It is used for the treatment of dyspepsia and chronic heart failure according to the Chinese Pharmacopoeia (2020). However, fungal contamination in Crataegi Fructus affects its quality and safety, thus preventing its global promotion. In this study, we comprehensively studied the fungal community in processed products of Crataegi Fructus by high-throughput sequencing. A total of 21 Crataegi Fructus samples were collected from five provinces in China, and the samples were divided into five groups based on collection areas, as well as into three groups based on processing methods. We then targeted the internal transcribed spacer 2 sequence through the Illumina Miseq PE300 platform to investigate fungal composition and diversity. Results showed that all 21 samples were detected with fungal contamination, and Ascomycota was dominant at the phylum level. In the groups based on collection areas, Dothideomycetes, Pleosporaceae, and Alternaria were dominant at the class, family, and genus levels, respectively. In the groups based on processing methods, Dothideomycetes, Aspergillaceae, and Alternaria were the most abundant at the class, family, and genus levels, respectively. Differences in fungal communities between various groups were also observed. Furthermore, a total of 115 species were identified, among which seven were potential toxigenic, namely, Trichothecium roseum, Alternaria tenuissima, Aspergillus carbonarius, Penicillium brevicompactum, Aspergillus fumigatus, Rhizopus microspores, and Pichia fermentans. In conclusion, this study reveals great fungal richness and diversity of Crataegi Fructus, providing references for the prevention and control of fungal contamination of Crataegi Fructus in practical production.

Identification and poisoning diagnosis of Aconitum materials using a genus-specific nucleotide signature.

Aconitum genus generally contains hypertoxic alkaloids. Poisoning incidents due to the improper ingestion of Aconitum materials frequently occur around the world. DNA barcoding is considered as a powerful tool for species identification, but complete sequences of conventional DNA barcodes are sometimes unattainable from food and highly processed products due to severe DNA degradation. Therefore, a shorter molecular marker will be more profitable for the authentication and poisoning diagnosis of Aconitum materials. In this study, 1246 psbA-trnH sequences and chloroplast genomes representing 183 taxa of Aconitum were collected, and a 23-bp nucleotide signature unique to Aconitum genus (5'-TATATGAGTCATTGAAGTTGCAG-3') was developed. The nucleotide signature was conserved and universal within Aconitum while divergent among other genera. The specific molecular signature was then successfully applied to the detection of processed Aconitum ingredients. To further evaluate the application potential of nucleotide signature in completely unknown mixture samples, boiled food mixtures, containing different ratios of Aconitum materials, were sequenced by high-throughput sequencing technology. The results showed that the nucleotide signature sequence could be directly extracted from raw sequencing data, even at a low DNA concentration of 0.2 ng/µl. Consequently, the 23-bp genus-specific nucleotide signature represents a significant step forward in the use of DNA barcoding to identify processed samples and food mixtures with degraded DNA. This study undoubtedly provides a new perspective and strong support for the identification and detection of Aconitum-containing products, which can be further introduced to the diagnosis of food poisoning.

Detection of Adulteration and Pesticide Residues in Chinese Patent Medicine Qipi Pill Using KASP Technology and GC-MS/MS.

Chinese patent medicines (CPMs) are of great value for the prevention and treatment of diseases. However, adulterants and pesticide residues in CPMs have become the "bottleneck" impeding the globalization of traditional Chinese medicine. In this study, 12 batches of commercially available Qipi pill (a famous CPM recorded in Chinese Pharmacopeia) from different manufacturers were investigated to evaluate their authenticity and quality safety. Considering the severely degraded DNA in CPMs, kompetitive allele specific PCR (KASP) technology combined with DNA mini-barcodes was proposed for the quality regulation of a large number of products in CPM market. The residues of four kinds of pesticides including pentachloronitrobenzene (PCNB), hexachlorocyclohexane (HCH), aldrin, and dichlorodiphenyltrichloroethane (DDT) were quantified using gas chromatography and tandem mass spectrometry (GC-MS/MS). The results indicated that in two of the 12 batches of Qipi pill, the main herbal ingredient Panax ginseng was completely substituted by P. quinquefolius, and one sample was partially adulterated with P. quinquefolius. The PCNB residue was detected in 11 batches of Qipi pill, ranging from 0.11 to 0.46 mg/kg, and the prohibited pesticide HCH was present in four samples. Both adulteration and banned pesticides were found in two CPMs. This study suggests that KASP technology combined with DNA mini-barcodes can be used for the quality supervision of large sample size CPMs with higher efficiency but lower cost. Our findings also provide the insight that pesticide residues in CPMs should be paid more attention in the future.

Plastid genome data provide new insights into the phylogeny and evolution of the genus Epimedium.

INTRODUCTION: Epimedium L., the largest herbaceous genus of Berberidaceae, is one of the most taxonomically difficult representatives. The classification and phylogenetic relationships within Epimedium are controversial and unresolved. OBJECTIVES: For the first time, we systematically studied the phylogeny and evolution of Epimedium based on plastid genome (plastome) data for better understanding this enigmatic genus. METHODS: We explored the molecular phylogeny, assessed the infrageneric classification, estimated the divergence times, and inferred the ancestral states for flower traits of Epimedium based on 45 plastomes from 32 species. RESULTS: The Epimedium plastome length ranged from 156,635 bp to 159,956 bp. Four types of plastome organization with different inverted repeat boundary changes were identified. Phylogenetic analysis revealed a strong support for the sister relationship of sect. Macroceras and sect. Diphyllon but did not provide a distinct route for petal evolution in sect. Diphyllon. Disharmony between phylogenetic relationships and traditional classification of sect. Diphyllon was observed. Results from divergence time analysis showed that Epimedium diverged in the early Pleistocene (~2.11 Ma, 95% HPD = 1.88-2.35 Ma). Ancestral character state reconstructions indicated transitions from long spur (large-flowered group) to other petal types (small-flowered group) in Epimedium. CONCLUSION: These findings provide new insights into the relationships among Epimedium species and pave the way for better elucidation of the classification and evolution of this genus.

Cross-frequency and iso-frequency estimation of functional corticomuscular coupling after stroke.

Functional corticomuscular coupling (FCMC) between the brain and muscles has been used for motor function assessment after stroke. Two types, iso-frequency coupling (IFC) and cross-frequency coupling (CFC), are existed in sensory-motor system for healthy people. However, in stroke, only a few studies focused on IFC between electroencephalogram (EEG) and electromyogram (EMG) signals, and no CFC studies have been found. Considering the intrinsic complexity and rhythmicity of the biological system, we first used the wavelet package transformation (WPT) to decompose the EEG and EMG signals into several subsignals with different frequency bands, and then applied transfer entropy (TE) to analyze the IFC and CFC relationship between each pair-wise subsignal. In this study, eight stroke patients and eight healthy people were enrolled. Results showed that both IFC and CFC still existed in stroke patients (EEG → EMG: 1:1, 3:2, 2:1; EMG → EEG: 1:1, 2:1, 2:3, 3:1). Compared with the stroke-unaffected side and healthy controls, the stroke-affected side yielded lower alpha, beta and gamma synchronization (IFC: beta; CFC: alpha, beta and gamma). Further analysis indicated that stroke patients yielded no significant difference of the FCMC between EEG → EMG and EMG → EEG directions. Our study indicated that alpha and beta bands were essential to concentrating and maintaining the motor capacities, and provided a new insight in understanding the propagation and function in the sensory-motor system.

Multiscale multivariate transfer entropy and application to functional corticocortical coupling.

Objective. Complex biological systems consist of multi-level mechanism in terms of within- and cross-subsystems correlations, and they are primarily manifested in terms of connectivity, multiscale properties, and nonlinearity. Existing studies have each only explored one aspect of the functional corticocortical coupling (FCCC), which has some limitations in portraying the complexity of multivariable systems. The present study investigated the direct interactions of brain networks at multiple time scales.Approach. We extended the multivariate transfer entropy (MuTE) method and proposed a novel method, named multiscale multivariate transfer entropy (MSMVTE), to explore the direct interactions of brain networks across multiple time scale. To verify this aim, we introduced three simulation models and compared them with multiscale transfer entropy (MSTE) and MuTE methods. We then applied MSMVTE method to analyze FCCC during a unilateral right-hand steady-state force task.Main results. Simulation results showed that the MSMVTE method, compared with MSTE and MuTE methods, better detected direct interactions and avoid the spurious effects of indirect relationships. Further analysis of experimental data showed that the connectivity from left premotor/sensorimotor cortex to right premotor/sensorimotor cortex was significantly higher than that of opposite directionality. Furthermore, the connectivities from central motor areas to both sides of premotor/sensorimotor areas were higher than those of opposite directionalities. Additionally, the maximum coupling strength was found to occur at a specific scale (3-10).Significance. Simulation results confirmed the effectiveness of the MSMVTE method to describe direct relationships and multiscale characteristics in complex systems. The enhancement of FCCC reflects the interaction of more extended activation in cortical motor regions. Additionally, the neurodynamic process of brain depends not only on emergent behavior at small scales, but also on the constraining effects of the activity at large scales. Taken together, our findings provide a basis for better understanding dynamics in brain networks.

Assessment of the Microbiome and Potential Aflatoxin Associated With the Medicinal Herb Platycladus orientalis.

Platycladi Semen, which is derived from the dried ripe seed of Platycladus orientalis, has been used for the treatment of insomnia and constipation in China for 2000 years. However, it is susceptible to fungal and aflatoxin contamination under proper humidity and temperature during storage. Although aflatoxin contamination in Platycladi Semen has been reported preliminarily, few studies have been conducted on fungal infection and aflatoxin contamination simultaneously. Thus, this work aims to provide an in-depth understanding of fungal contamination in Platycladi Semen, and information on aflatoxin contamination. We focused on a comparison of the difference in fungal diversity between aflatoxin-contaminated and aflatoxin-free Platycladi Semen samples. First, aflatoxin levels in 11 Platycladi Semen samples, which were collected from local herbal markets in Shandong, Anhui, and Hebei provinces throughout China, were determined by IAC-HPLC-FLD, and positive confirmation of detected samples was performed by LC-MS/MS. The samples were divided into two groups, based on production or non-production of aflatoxin. We then used the Illumina MiSeq PE250 platform, and targeted the internal transcribed spacer two sequences to analyze the diversity and composition of the fungal microbiome, as well as to assess the presence of potential mycotoxin-producing fungi. Results showed that five samples were contaminated with aflatoxins, one of which exceeded the legal limits of Chinese Pharmacopeia Commission (2015). At the phylum level, the Ascomycota was the most dominant in all tested samples, with a relative abundance of 83.04-99.46%. Aspergillus (27.88-97.28%), Xerochrysium (0-28.49%), and Xeromyces (0-22.24%) were the three predominant genera. Furthermore, differences in fungal composition between the aflatoxin-contaminated and aflatoxin-free groups, as well as between different provinces were observed. A total of 74 species were identified, and four potential mycotoxin-producing fungi were detected in all samples, namely Aspergillus flavus, Aspergillus fumigatus, Fusarium poae, and Penicillium steckii. In conclusion, we report the great diversity of fungi associated with Platycladi Semen, highlight the risk to consumers of ingesting potent aflatoxin, and provide a reference for the safe application and quality improvement of Platycladi Semen.