Integration of Untargeted Metabolomics and Object-Oriented Data-Processing Protocols to Characterize Acerola Powder Composition as Functional Food Ingredient.

Acerola powder has been experiencing a surge in demand as a functional food ingredient, particularly due to its usage in vitamin C supplements. However, limited research has been conducted on its other bioactive compounds. In this study, we employed metabolomics and object-oriented data-processing protocols to comprehensively characterize acerola powder. To ensure maximum coverage of metabolomics, we selected a 50% methanol aqueous solution as the extraction solvent and utilized the HSS T3 column for chromatography analysis. Through this approach, we successfully identified a total of 175 compounds in acerola powder, encompassing amino acids and peptides, polyphenols, organic acids, and various other compounds. Additionally, we measured the total phenolic content (TPC) and assessed the antioxidant activity of acerola powder. Furthermore, we analyzed the differential composition of acerola fruit and juice powder, identifying polyphenols and lipids as primary markers in fruit powder, while peptides emerged as key markers in juice powder. Notably, two specific peptides, Thr-Trp and Val-Tyr, were identified as antioxidant peptides. Overall, our study provides novel composition data for acerola powder, shedding light on its potential as a functional food ingredient. These findings contribute to the development and utilization of acerola powder in the formulation of functional food products.

X-Ray2EM: Uncertainty-Aware Cross-Modality Image Reconstruction from X-Ray to Electron Microscopy in Connectomics.

Comprehensive, synapse-resolution imaging of the brain will be crucial for understanding neuronal computations and function. In connectomics, this has been the sole purview of volume electron microscopy (EM), which entails an excruciatingly difficult process because it requires cutting tissue into many thin, fragile slices that then need to be imaged, aligned, and reconstructed. Unlike EM, hard X-ray imaging is compatible with thick tissues, eliminating the need for thin sectioning, and delivering fast acquisition, intrinsic alignment, and isotropic resolution. Unfortunately, current state-of-the-art X-ray microscopy provides much lower resolution, to the extent that segmenting membranes is very challenging. We propose an uncertainty-aware 3D reconstruction model that translates X-ray images to EM-like images with enhanced membrane segmentation quality, showing its potential for developing simpler, faster, and more accurate X-ray based connectomics pipelines.

Strategy for anthocyanins production: From efficient green extraction to novel microbial biosynthesis.

Anthocyanins are widely distributed in nature and exhibit brilliant colors and multiple health-promoting effects; therefore, they are extensively incorporated into foods, pharmaceuticals, and cosmetic industries. Anthocyanins have been traditionally produced by plant extraction, which is characterized by high expenditure, low production rates, and rather complex processes, and hence cannot meet the increasing market demand. In addition, the emerging environmental issues resulting from traditional solvent extraction technologies necessitate a more efficient and eco-friendly alternative strategy for producing anthocyanins. This review summarizes the efficient approach for green extraction and introduces a novel strategy for microbial biosynthesis of anthocyanins, emphasizing the technological changes in production.

Cloning and functional verification of PhAEP gene, a key enzyme for biosynthesis of heterophyllin A in Pseudostellaria heterophylla / 中国中药杂志

This paper aimed to study the role of asparagine endopeptidase(AEP) gene in the biosynthesis mechanism of cyclic peptide compounds in Pseudostellaria heterophylla. The transcriptome database of P. heterophylla was systematically mined and screened, and an AEP gene, tentatively named PhAEP, was successfully cloned. The heterologous function verification by Nicotiana benthamiana showed that the expression of the gene played a role in the biosynthesis of heterophyllin A in P. heterophylla. Bioinformatics analysis showed that the cDNA of PhAEP was 1 488 bp in length, encoding 495 amino acids with a molecular weight of 54.72 kDa. The phylogenetic tree showed that the amino acid sequence encoded by PhAEP was highly similar to that of Butelase-1 in Clitoria ternatea, reaching 80%. The sequence homology and cyclase active site analysis revealed that the PhAEP enzyme may specifically hydrolyse the C-terminal Asn/Asp(Asx) site of the core peptide in the HA linear precursor peptide of P. heterophylla, thereby participating in the ring formation of the linear precursor peptide. The results of real-time quantitative polymerase chain reaction(RT-qPCR) showed that the expression level of PhAEP was the highest in fruits, followed by in roots, and the lowest in leaves. The heterophyllin A of P. heterophylla was detected in N. benthamiana that co-expressed PrePhHA and PhAEP genes instantaneously. In this study, the PhAEP gene, a key enzyme in the biosynthesis of heterophyllin A in P. heterophylla, has been successfully cloned, which lays a foundation for further analysis of the molecular mechanism of PhAEP enzyme in the biosynthesis of heterophyllin A in P. heterophylla and has important significance for the study of synthetic biology of cyclic peptide compounds in P. heterophylla.

Skin transcriptome reveals the periodic changes in genes underlying cashmere (ground hair) follicle transition in cashmere goats.

BACKGROUND: Cashmere goats make an outstanding contribution to the livestock textile industry and their cashmere is famous for its slenderness and softness and has been extensively studied. However, there are few reports on the molecular regulatory mechanisms of the secondary hair follicle growth cycle in cashmere goats. In order to explore the regular transition through the follicle cycle and the role of key genes in this cycle, we used a transcriptome sequencing technique to sequence the skin of Inner Mongolian cashmere goats during different months. We analyzed the variation and difference in genes throughout the whole hair follicle cycle. We then verified the regulatory mechanism of the cashmere goat secondary hair follicle growth cycle using fluorescence quantitative PCR. RESULTS: The growth cycle of cashmere hair could be divided into three distinct periods: a growth period (March-September), a regression period (September-December), and a resting period (December-March). The results of differential gene analyses showed that March was the most significant month. Cluster analysis of gene expression throughout the whole growth cycle further supported the key nodes of the three periods of cashmere growth, and the differential gene expression of keratin corresponding to the ground haircashmere growth cycle further supported the results from tissue slices. Quantitative fluorescence analysis showed that KAP3-1, KRTAP 8-1, and KRTAP 24-1 genes had close positive correlation with the cashmere growth cycle, and their regulation was consistent with the growth cycle of cashmere. CONCLUSION: The growth cycle of cashmere cashmere could be divided into three distinct periods: a growth period (March-September), a regression period (September-December) and a resting period (December-March). March was considered to be the beginning of the cycle. KAP and KRTAP showed close positive correlation with the growth cycle of secondary hair follicle cashmere growth, and their regulation was consistent with the cashmere growth cycle. But hair follicle development-related genes are expressed earlier than cashmere growth, indicating that cycle regulation could alter the temporal growth of cashmere. This study laid a theoretical foundation for the study of the cashmere development cycle and provided evidence for key genes during transition through the cashmere cycle. Our study provides a theoretical basis for cashmere goat breeding.

Effects of Low Temperature at Booting Stage on Sucrose Metabolism and Endogenous Hormone Contents in Winter Wheat Spikelet.

Low spring temperatures often occur during the winter wheat booting stage, when the young ears are very sensitive to cold. In this study, we used two wheat varieties differing in cold sensitivity (sensitive variety Yangmai 18 and tolerant variety Yannong 19) to examine the effect of low temperature on wheat grain number at booting stage. Low temperature stress was simulated in an artificial climate chamber at 4°C for 60 h in 2016 and at 2, 0, or -2°C for 24 h in morphological assays, showing that the development of wheat spikelets was inhibited and floret growth was delayed following low temperature stress. However, an increase in the sucrose content of young panicles was also observed, and the activity of enzymes involved in sucrose metabolism was dynamically altered. Sucrose phosphate synthase activity was enhanced, and sucrose synthase activity significantly increased after treatment at 4 and 2°C, respectively. However, activities of sucrose synthase and invertase decreased with a reduction in temperature. Gene expression assays further revealed downregulation of TaSuS1 expression and upregulation of TaSuS2, while expression of CWINV was inhibited. Moreover, phytohormone content assays showed an increase in the content of abscisic acid in young wheat ears, but a decrease in the content of auxin and gibberellins. The grain number per spike and 1000-grain weight also showed a downward trend following low temperature stress. Overall, these findings suggest that low temperature at booting induces abscisic acid accumulation in winter wheat, altering the activity of the enzymes involved in sucrose metabolism, which leads to an accumulation of sucrose in the young ears, thereby having a negative effect on wheat production.

Molecular engineering mechanism of organic photoactive layer by alkyl chains, 4-butoxyphenyl and cyanogroup.

The three organic dye molecules (JY31, JY32 and JY33) were applied to the photoactive layer in solar cells. Photophysical and photochemical characteristic have been investigated with natural bond orbital (NBO), frontier molecular orbital, ionization potentials, electron affinities, absorption properties, reorganization energies, static first hyperpolarizability, emission characteristics, IR spectra, charge density difference; the influence of alkyl chains and 4-butoxyphenyl on properties were revealed; Subsequently, three new molecules JY33-1, JY33-2 and JY33-3 were designed by inserting the electron withdrawing group -CN into the acceptor part of JY33 in order to understand molecular engineering mechanism. The results show that the three original molecules have relatively high molar extinction coefficients, and the molecule of JY33 with a 4-butoxyphenyl group enables a bathochromic shift in absorption spectrum and is beneficial to improve the hole transport, injection capacity and ICT properties as well as better energy levels matching. The current study provides an effective channel for manipulating performance in materials design of solar cells.

Design, Electron Transfer Process, and Opto-Electronic Property of Solar Cell Using Triphenylamine-Based D-π-A Architectures.

A series of D-π-A type dyes were designed based on the experimentally synthesized A1 by introducing different functional groups on the donor and π-spacer, and the optical and electrical properties were calculated by using density functional theory (DFT) and time-dependent DFT (TD-DFT). P1⁻P6 present highest light harvesting efficiency (LHE), driving force of electron injection ( Δ G i n j e c t ), reorganization energy ( Δ G r e g ) and e V O C . These critical parameters have a close relationship with the short-circuit current density ( J S C ) and open-circuit photovoltage ( V O C ), and lead to P1⁻P6 will exhibit higher efficiency. D4 also exhibit superior properties in the driving force of electron injection ( Δ G i n j e c t ), reorganization energy ( Δ G r e g ), which will lead to a higher short-circuit current density ( J S C ). We hope that these results will be helpful for experiments to synthesize new and highly efficient dyes.

Incremental diagnostic value of neck vessel wall imaging technique with T1?weighted three?dimensional variable?flip?angle turbo spin?echo before revascularization in patients with carotid atherosclerotic disease / 中华放射学杂志

Objective To determine the feasibility of neck vessel wall imaging technology with three?dimensional variable?flip?angle turbo spin?echo (3D T1w?SPACE) for the detection of carotid atherosclerotic disease before revascularization. Methods Thirty?one patients who underwent carotid endarterectomy (CEA) and fifty?three patients who underwent carotid stenting (CAS) were enrolled prospectively. Neck vessel wall imaging examination were performed in all patients whilecarotid artery DSA were performed in all CAS patients. Quantitative measurements including stenosis, lesion length, and the presence or absence of plaque ulceration obtained with 3D T1w?SPACE and DSA were independently determined. And images of the 3D T1w?SPACE were compared with corresponding histology to identify major plaque components including intraplaque hemorrhage (IPH), lipid rich necrotic core (LRNC), and calcification (CA). The consistency rate, sensitivity, specificity, positive predictive value and negative predictive value were used to assess diagnostic value. Bland?Altman plots, intraclass correlation coefficient (ICC), and Cohen Kappa were determined. Results DSA was served as the reference standard. There was an excellent correlation between 3D T1w?SPACE and DSA images in measuring stenosis (r=0.984, P<0.01) and luminal stenosis [ICC=0.98 (95% confidence interval: 0.96-0.99)]. Bland?Altman plots showed that the two examinations were in good consistency in evaluating the extent of stenosis. Sensitivity (89.5%) and specificity (95.1%) was high in 3D T1w?SPACE images compared to DSA for the detection of ulcers. The consistency rate between 3D T1w?SPACE images and histological results for IPH, LRNC and CA detection were 85.7%, 82.1% and 92.9%, respectively. Sensitivity and specificity were 90.0% and 75.0% for IPH;83.3% and 80.0% for LRNC; 91.3% and 100.0% for CA respectively. However, lesion length measurements by using 3D T1w?SPACE were longer than those measured by using DSA (P<0.01).Conclusion Neck vessel wall imaging technology with 3D T1w?SPACE is a noninvasive and accurate technique for the diagnosis of carotid artery atherosclerotic disease before revascularization.

Investigation of Integrated Pharmacokinetics of Active Components in Moutan Cortex and Its Correlation with Pharmacodynamics / 中国实验方剂学杂志

Objective:To study on the correlation between integrated pharmacokinetics and pharmacodynamic effects of five active components(oxidized paeoniflorin,paeoniflorin,quercetin,gallic acid, paeonol) in Moutan Cortex. Method:Rats were divided into blank group,model group(syndrome of blood-heat and blood stasis) and drug-administered group.The concentration of five active components in serum were detected with UPLC-MS at different time points after being administrated ethanol extract of Moutan Cortex.The integrated concentrations were calculated according to area under the curve(AUC) self-defined weighting coefficiency.At the same time,the enzyme-linked immunosorbent assay(ELISA) was used to determine the contents of thromboxane B2(TXB2) and 6-keto-prostaglandin F1α(6-keto-PGF1α) in serum at different time points,and then correlation between pharmacodynamics and integrated pharmacokinetics of these five active ingredients was analyzed. Result:At different time points(0.083,0.25,0.5,0.75,1,2,3,4,6,8,10,12 h),the integrated plasma concentrations of these five active ingredients in Moutan Cortex(158.65,174.60,220.13,227.23,244.31,251.51,404.28,654.39,472.62,355.04, 231.56,199.40 mg·L-1) had a good correlation with concentration of TXB2(264.44,261.03,284.93,273.30,264.04, 278.90,274.83,303.58,260.03,264.78,264.40,256.62 μg·L-1) and value of TXB2/6-keto-PGFlα(4.50,4.47,3.66,3.37, 3.29,3.66,3.71,4.30,3.63,3.65,3.75,3.66). Conclusion:There is a good correlation between the dynamic changes in vivo of active components from Moutan Cortex and pharmacodynamic effects of activating blood circulation of this herb.

A fluidic circuit based, high-efficiency and large-scale single cell trap.

Single cell traps have important applications in biological cell manipulation and analysis. This paper describes a novel single cell trap design and device with a matrix of cell trap units inspired by an equivalent resistive electric circuit. This fluidic device follows the least flow resistance path principle of such devices allowing deterministic single cell trapping with high efficiency and flexibility for large scale cell patterning.

Study of a Microfluidic Chip Integrating Single Cell Trap and 3D Stable Rotation Manipulation.

Single cell manipulation technology has been widely applied in biological fields, such as cell injection/enucleation, cell physiological measurement, and cell imaging. Recently, a biochip platform with a novel configuration of electrodes for cell 3D rotation has been successfully developed by generating rotating electric fields. However, the rotation platform still has two major shortcomings that need to be improved. The primary problem is that there is no on-chip module to facilitate the placement of a single cell into the rotation chamber, which causes very low efficiency in experiment to manually pipette single 10-micron-scale cells into rotation position. Secondly, the cell in the chamber may suffer from unstable rotation, which includes gravity-induced sinking down to the chamber bottom or electric-force-induced on-plane movement. To solve the two problems, in this paper we propose a new microfluidic chip with manipulation capabilities of single cell trap and single cell 3D stable rotation, both on one chip. The new microfluidic chip consists of two parts. The top capture part is based on the least flow resistance principle and is used to capture a single cell and to transport it to the rotation chamber. The bottom rotation part is based on dielectrophoresis (DEP) and is used to 3D rotate the single cell in the rotation chamber with enhanced stability. The two parts are aligned and bonded together to form closed channels for microfluidic handling. Using COMSOL simulation and preliminary experiments, we have verified, in principle, the concept of on-chip single cell traps and 3D stable rotation, and identified key parameters for chip structures, microfluidic handling, and electrode configurations. The work has laid a solid foundation for on-going chip fabrication and experiment validation.

EFFECT OF INTASPINAL IMPLANTATION OF MICROGENE pSVPoMcat GENETICALLY MODIFIED SCHWANN CELLS ON SPINAL CORD INJURY REPAIR IN RATS / 解放军医学杂志

To observe the effect of intaspinal implantation of Schwann cells (SC) genetically modified with microgene pSVPoMcat on spinal cord injury (SCI) repair.120 SD rats were used to establish the hemisected spinal cord injury model at T 8 level,and they were divided randomly into three groups: genetically modified SC implantation group (group A),normal SC implantation group (group B) and control group without cell implantation (group C).One week after the operation ,combined behavioral score(CBS) and the cortical somatasensory evoked potential (GFAP) were measured and the expression of glial fibrillary acidic protein(GFAP) was examined by in situ hybridization and immunocytochemistry.Three months after the operation, all the rats were scanned with MRI and then were sacrificed.Neurofilament (NF) was examined with imunohistocytochemistry staining by using NF monoclonal antibody. Following were the results:(1) In group A,the number of cells expressed GFAP in injured sites was less than that in groups B and C.(2) MRI scanning showed that the SCI region almost recovered in group A but did not recover in group B.There was a malacie focus in SCI region in group C.This was corroborated by the NF staining.(3) The amplitudes of potential in the latent period in group A and B showed a tendency to recover,and it was consistent with CBS.The results suggested that the implantation of genetically modified SC with microgene pSVPoMcat could inhibit GFAP expression and promote the functional recovery of spinal cord injury in rats.