Exploring natural product biosynthesis in plants with mass spectrometry imaging.

The biosynthesis of natural products (NPs) is a complex dynamic spatial and temporal process that requires the collaboration of multiple disciplines to explore the underlying mechanisms. Mass spectrometry imaging (MSI) is a powerful technique for studying NPs due to its high molecular coverage and sensitivity without the need for labeling. To date, many analysts still use MSI primarily for visualizing the distribution of NPs in heterogeneous tissues, although studies have proved that it can provide crucial insights into the specialized spatial metabolic process of NPs. In this review we strive to bring awareness of the importance of MSI, and we advocate further exploitation of the spatial information obtained from MSI to establish metabolite-gene expression relationships.

Obesity-induced upregulation of miR-483-5p impairs the function and identity of pancreatic ß-cells.

AIM: To assess the expression and function of miR-483-5p in diabetic ß cells. METHODS: The expression of miR-483-5p was evaluated in the pancreatic islets of obesity mouse models by quantitative reverse transcription polymerase chain reaction. Dual-luciferase activity, and western blotting assays, were utilized for miR-483-5p target gene verification. Mice with ß cell-specific miR-483-5p downregulation were studied under metabolic stress (i.e. a high-fat diet) condition. Lineage tracing was used to determine ß-cell fate. RESULTS: miR-483-5p increased in the islets of obese mouse models. Expression levels of miR-483-5p were significantly upregulated with the treatment of high glucose and palmitate, in both MIN6 cells and mouse islets. Overexpression of miR-483-5p in ß cells results in impaired insulin secretion and ß-cell identity. Cell lineage-specific analyses revealed that miR-483-5p overexpression deactivated ß-cell identity genes (insulin, Pdx1 and MafA) and derepressed ß-cell dedifferentiation (Ngn3) genes. miR-483-5p downregulation in ß cells of high-fat diet-fed mice alleviated diabetes and improved glucose intolerance by enhancing insulin secretory capacity. These detrimental effects of miR-483-5p relied on its seed sequence recognition and repressed expression of its target genes Pdx1 and MafA, two crucial markers of ß-cell maturation. CONCLUSIONS: These findings indicate that the miR-483-5p-mediated reduction of mRNAs specifies ß-cell identity as a contributor to ß-cell dysfunction via the loss of cellular differentiation.

Reversing Metabolic Dysfunction-Associated Steatotic Liver Disease Promotes Healthy Pediatric Cardiovascular Structures: The PROC Study.

INTRODUCTION: We investigated the impact of metabolic dysfunction-associated steatotic liver disease (MASLD) on cardiovascular structure development in children. METHODS: We followed 1,356 children with the mean age of 6.6 years for 4.5 years in Beijing, China. We assessed the association of MASLD with cardiovascular structure (carotid intima-media thickness and left ventricular mass) outcomes at baseline and follow-up. RESULTS: Over follow-up, 59 children had persistent MASLD, 109 had incident MASLD (progression), and 35 had normalization of liver health. Children with MASLD normalization showed a significantly lower mean development in carotid intima-media thickness (0.161 vs 0.188 mm) and left ventricular mass (4.5 vs 12.4 g) than children with persistent MASLD. DISCUSSION: The control of MASLD was associated with improved cardiovascular structure development.

A multimodal pipeline for image correction and registration of mass spectrometry imaging with microscopy.

The integration of matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) and histology plays a pivotal role in advancing our understanding of complex heterogeneous tissues, which provides a comprehensive description of biological tissue with both wide molecule coverage and high lateral resolution. Herein, we proposed a novel strategy for the correction and registration of MALDI MSI data with hematoxylin & eosin (H&E) staining images. To overcome the challenges of discrepancies in spatial resolution towards the unification of the two imaging modalities, a deep learning-based interpolation algorithm for MALDI MSI data was constructed, which enables spatial coherence and the following orientation matching between images. Coupled with the affine transformation (AT) and the subsequent moving least squares algorithm, the two types of images from one rat brain tissue section were aligned automatically with high accuracy. Moreover, we demonstrated the practicality of the developed pipeline by projecting it to a rat cerebral ischemia-reperfusion injury model, which would help decipher the link between molecular metabolism and pathological interpretation towards microregion. This new approach offers the chance for other types of bioimaging to boost the field of multimodal image fusion.

Transcriptomic evidence of hypothalamus for maternal fructose exposure induced offspring hypertension through AT1R/TLR4 pathway.

Maternal fructose exposure during pregnancy and lactation has been shown to contribute to hypertension in offspring, with long-term effects on hypothalamus development. However, the underlying mechanisms remain unclear. In this study, we used the tail-cuff method to evaluate the effects of maternal fructose drinking exposure on offspring blood pressure levels at postpartum day 21 (PND21) and postpartum day 60 (PND60). We employed Oxford Nanopore Technologies (ONT) full-length RNA sequencing to investigate the developmental programming of the PND60 offspring's hypothalamus and confirmed the presence of the AT1R/TLR4 pathway using western blot and immunofluorescence. Our findings demonstrated that maternal fructose exposure significantly increased blood pressure in PND60 offspring but not in PND21 offspring. Additionally, we observed transcriptome-wide alterations in the hypothalamus of PND60 offspring following maternal fructose exposure. Overall, our study provides evidence that maternal fructose exposure during pregnancy and lactation may alter the transcriptome-wide of offspring hypothalamus and activate the AT1R/TLR4 pathway, leading to hypertension. These findings may have important implications for the prevention and treatment of hypertension-related diseases in offspring exposed to excessive fructose during pregnancy and lactation.

Alternative splicing affects synapses in the hippocampus of offspring after maternal fructose exposure during gestation and lactation.

Increased fructose over-intake is a global issue. Maternal fructose exposure during gestation and lactation can impair brain development in offspring. However, the effect on synapses is still unknown. For the diversification of RNA and biological functions, alternative splicing (AS) and alternative polyadenylation (APA) are essential. We constructed a maternal high-fructose diet model by administering 13% and 40% fructose water. The student's t-test analyzed the results of RT-qPCR. All other results were analyzed by one-way analysis of variance. The animal behavior experiment results revealed that conditioning and associative memory had been damaged. The proteins that form synapses were consistently low-expressed. In addition, compared with the control group, the Oxford Nanopore Technologies platform's full-length RNA-sequencing identified 298 different spliced genes (DSGs) and 51 differentially expressed alternative splicing (DEAS) genes in the 13% fructose group. 313 DSGs and 74 DEAS genes were in the 40% fructose group. Enrichment analysis based on these altered genes revealed some enlightening items and pathways. Our findings demonstrated the transcriptome mechanism underlying maternal fructose exposure during gestation and lactation and impaired synapse function during the transcripts' editing.

Effects of Maternal High-Fructose Diet on Long Non-Coding RNAs and Anxiety-like Behaviors in Offspring.

Increased fructose intake is an international issue. A maternal high-fructose diet during gestation and lactation could affect nervous system development in offspring. Long non-coding RNA (lncRNA) plays an important role in brain biology. However, the mechanism whereby maternal high-fructose diets influence offspring brain development by affecting lncRNAs is still unclear. Here, we administered 13% and 40% fructose water to establish a maternal high-fructose diet model during gestation and lactation. To determine lncRNAs and their target genes, full-length RNA sequencing was performed using the Oxford Nanopore Technologies platform, and 882 lncRNAs were identified. Moreover, the 13% fructose group and the 40% fructose group had differentially expressed lncRNA genes compared with the control group. Enrichment analyses and co-expression analyses were performed to investigate the changes in biological function. Furthermore, enrichment analyses, behavioral science experiments, and molecular biology experiments all indicated that the fructose group offspring showed anxiety-like behaviors. In summary, this study provides insight into the molecular mechanisms underlying maternal high-fructose diet-induced lncRNA expression and co-expression of lncRNA and mRNA.

Non-suicidal self-injury: A bibliometrics study and visualization analysis from 2002 to 2022.

Objective: To overview the research actuality and offer the hotspots and cutting-edge issues in the field of Non-suicidal Self-injury (NSSI) by using bibliometric analysis. Materials and methods: Publications related to NSSI from 2002 to 2022 were extracted from the Web of Science Core Collection (WoSCC) database. CiteSpace V 6.1.R2 and VOSviewer 1.6.18 were used to visually analyzed institutions, countries, journals, authors, references, and keywords in research on NSSI. Results: A total of 799 studies about NSSI were analyzed via CiteSpace and VOSviewer. The number of annual publications related to NSSI is fluctuating growth. The USA and Harvard University are the most productive country and institutions. In the case of journals and co-cited journals, Psychiatry Research are the most productive journal and also ranked highest among co-cited journals. Furthermore, Michael Kaess has published the most publications, and Matthew K. Nock is the most cited author. An article published by Swannell SV et al. shows the highest citation counts. After analysis, the most common keywords are harm, adolescents and prevalence. The gender difference, diagnosis, and dysregulation are frontier areas of NSSI research. Conclusion: This study analyzed the research of NSSI from multiple perspectives, and provides valuable information for researchers to capture the current status, hot spots, and frontier trends of NSSI.

Spatial segmentation of mass spectrometry imaging data featuring selected principal components.

Spatial segmentation aims to find homogeneous/heterogeneous subgroups of spectra or ion images in mass spectrometry imaging (MSI) data. The maps it generated inform researchers of vital characteristics of the data and thus provide the basis for strategizing further biological analysis. Dimensional reduction and clustering are two basic steps of segmentation. Due to the variations in the quality, resolution, density of spectral information, and sizes, not all datasets could be segmented ideally with combinations of different dimensional reduction and clustering algorithms. Here, we proposed a segmentation pipeline that utilized pattern compression by principal component analysis (PCA) and represented by principal components. Instead of preprocessed or raw MSI data, normalized principal components were used for the segmentation process. Multiple datasets of rat brains and mouse kidneys were tested, and the proposed segmentation pipeline presented the obvious advantage of easy-to-use and can be readily intergraded with other existing innovative pipelines.

Learning and memory impairment and transcriptomic profile in hippocampus of offspring after maternal fructose exposure during gestation and lactation.

Increased fructose intake is a global issue, especially in mothers. Maternal fructose exposure during gestation and lactation can affect learning and memory in offspring; however, the detailed mechanism is still unknown. The hippocampus is a mind locale liable for learning and memory. Here, we established a maternal high-fructose diet model by administering 13% and 40% fructose water, applied the Morris Water Maze test on postnatal day 60 offspring, and performed full-length RNA sequencing using the Oxford Nanopore Technologies platform to explore the changes in gene expression in the hippocampus. The results showed that learning and memory in offspring were negatively affected. Compared with the control group, 369 differentially expressed transcripts (DETs) were identified in the 13% fructose group, and 501 DETs were identified in the 40% fructose group. Gene Ontology enriched term and Kyoto Encyclopedia of Genes and Genomes enriched pathway analyses identified several terms and pathways related to brain development and cognitive function. Furthermore, we confirmed that the Wnt/ß-catenin signaling pathway was down-regulated and neuron degeneration was enhanced. In summary, our results indicate that maternal fructose exposure during gestation and lactation can impair learning and memory in offspring and affect brain function at the transcriptome level.

Mass spectrometry imaging and its potential in food microbiology.

Mass spectrometry imaging (MSI) is a tool capable of simultaneously providing in situ untargeted chemical information as well as the spatial distribution of vast molecular species with high efficiency. Over decades, this technique has been proven quite solid in many life science disciplines including microbiology, yet few food microbiologists have been using it. This review briefly introduces MSI technology and covers some interesting topics regarding sample preparations. Successful applications of MSI in disciplines similar to food microbiology will be described and perspectives of how MSI could benefit food microbiology and how it may be challenged will be given. This review aims to draw more attention from food microbiologists to this developing technology and subsequently inspire more actual applications.

Development and Pilot Testing a Self-Reported Pediatric PROMIS App for Young Children Aged 5-7 Years.

PURPOSE: The aims of this study are threefold. Firstly, Using the state of science PROMIS (Patient-Reported Outcomes Measurement Information System) methods to develop a smartphone application to monitor the emotional distress for young children aged 5-7 years old; Secondly, to test the usability of this application; and thirdly, to determine the level of agreement between reports by parents and young children's self-report. DESIGN AND METHODS: A multidisciplinary research team, made up of senior pediatric nurses and doctors, software engineers' team, and pediatric health researchers worked together to develop this application. Three phases of stakeholders and user studies were conducted. Phase 1 focused on prototype development; Phase 2 involved cognitive interview and usability testing; Phases 3 focused on the pilot testing of this application. RESULTS: We included the original parent proxy reporting version of Patient Reported Outcome Measurement Information System-emotional distress in the application, as well as self-reporting animated version for young children. After many rounds of modification, all participants felt that this application was easy to use and the animated items were easy to understand for young children aged 5-7 years. Correlations between parents-children reports are significant and moderate, parents underestimated child depression, and overestimated child anger and anxiety compared to child self-report. CONCLUSIONS: This smartphone application and its Web-based administration portal demonstrate good usability and are well accepted by young children aged 5-7 years, which can be used to promote young children's participation when reporting or assessing symptoms of young pediatric patients. PRACTICE IMPLICATIONS: Parent reports cannot be substituted for child reports and evaluations of pediatric patients' perspectives regarding treatment outcomes should be included in pediatric clinic. This animated application can be used as a smart measurement to investigate the symptoms for young children aged 5-7 years, so as to amplify young children's voice in clinical care.

Quantification of polyglutamyl 5-methyltetrahydrofolate, monoglutamyl folate vitamers, and total folates in different berries and berry juice by UHPLC-MS/MS.

Edible berries are good sources of several phytochemicals. However, the native folate levels in berries are not well known. The structure of native folates contains polyglutamyl chains, which reportedly jeopardize the bioavailability of native folates; further γ-glutamyl hydrolase (GGH) can deglutamylate polyglutamyl chains. In this study we use a validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to determine the distribution of polyglutamyl 5-methyltetrahydrofolate (5-CH3THF-Glun), folate vitamers, and total folates in different berries and to monitor changes in their concentration during processing of berries into juice. The pre-boiling treatment was optimized during extraction to stabilize the native polyglutamyl folates profile and facilitate folate extraction, which can replace the traditional di-enzyme treatment process. Additionally, the efficiency of commercially available human recombinant GGH was tested and it was found that a 10 µg GGH/mL extract at a pH of 6 could completely deconjugate polyglutamyl folates into monoglutamyl folates when incubated for 30 min. Pure human recombinant GGH with a higher catalytic efficiency and stable enzymatic properties was better than traditional folate conjugase for this purpose. From experimental analysis, it could be inferred that strawberries and blackberries contained the highest amount of total folates (93-118 µg/100 g), while the total folate contents in blueberries were the lowest. Most of the investigated berries are good to excellent folate sources. This study is the first time that 5-CH3THF-Glun and distribution of folate vitamers in various berries are quantitated. Further, this is the first study to show the application of recombinant pure GGH for the deconjugation of polyglutamyl folates for folate vitamers and total folates analysis.

High pressure processing and post-high pressure storage induce the change of polyglutamyl folate and total folate from different legumes.

The aim of this work was to study the effect of high pressure processing (HPP) and post-HPP cold storage on the distribution of polyglutamyl and monoglutamyl folate and the absolute concentration of total folate in green beans, yardlong beans and winged beans using a validated ultra-high performance liquid chromatography-tandem mass spectrometry method. The results showed that HPP led to the deglutamylation of polyglutamyl folate to monoglutamyl folate in all of the investigated beans. The degree of deglutamylation was increased with enhancing processing pressure and extending holding time. During HPP, significant loss of total folate was observed under 600 MPa/10 min treatment. Uniquely 300 MPa/5 min and 450 MPa/5 min could significantly release more folate from yardlong beans and green beans matrix. During the following cold-storage, the deglutamylation keep progressing. For those untreated beans, no significant deglutamylation and total folate loss was observed during cold storage for yardlong beans and green beans while there is slight change for the total folate in winged beans. For those HPP treated beans, total folate loss followed the first order kinetics over the storage. The rate constant of degradation was positively proportional to the applied pressure, holding time and the proportion of monoglutamyl folate. This research provided a reference for understanding the deglutamylation of polyglutamyl folate and folate loss during HPP treatment and further shelf life.

Quantification of Total Folate, Folate Species and Polyglutamyl Folate Distribution in Winged Beans (Psophocarus tetragonolobus (L) DC) from Different Cultivars and Growth Stages by Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry.

Winged beans are an important natural source of some micronutrients. This paper presents the first complete characterization of folate derivatives including polyglutamyl 5-methyltetrahydrofolate (5-CH3-H4PteGlun), folate species and total folate accumulating in pods and immature seeds of winged beans from 9 cultivars and 7 growth stages. 5-CH3-H4PteGlun and folate species were determined with a UHPLC-MS/MS method. Accurate determination of 5-CH3-H4PteGlun and folate species was optimized and validated according to EMA guidelines including method selectivity, sensitivity, linearity, accuracy, precision, matrix effect and carry-over. The level of total folate is in the range of 73-200 µg/100 g in the pods and 33-61 µg/100 g in the immature seeds. The predominant folate species in winged beans is 5-CH3-H4PteGlu1. 5-CH3-H4PteGlu5 is the major polyglutamyl folate derivative. The level of total folate is increased about 4 fold with advancing maturity. For pods, the chain length is increased with growth which shifts from 5-CH3-H4PteGlu1 in the early stage to 5-CH3-H4PteGlu5 and 5-CH3-H4PteGlu6 in the 7th stage. Our findings demonstrate that winged beans are good source of folate. The validated UHPLC-MS/MS method allows for the determination of 5-CH3-H4PteGlun and folate species from other vegetable matrices.

Gender differences in reward and punishment for monetary and social feedback in children: An ERP study.

Gender differences in feedback processing have been observed among adolescents and adults through event-related potentials. However, information on whether and how this feedback processing is affected by feedback valence, feedback type, and individual sensitivity in reward/punishment among children remains minimal. In this study, we used a guessing game task coupled with electroencephalography to investigate gender differences in feedback processing, in which feedback to reward and punishment was presented in the context of monetary and social conditions. Results showed that boys were less likely to switch their response after punishment, had generally less feedback-related negativity (FRN) amplitude, and longer FRN latency in monetary and punishment conditions than girls. Moreover, FRN for monetary punishment, which is related to individual difference in reward sensitivity, was observed only in girls. The study provides gender-specific evidence for the neural processing of feedback, which may offer educational guidance for appropriate feedback for girls and boys.

Social competition factor influences the neural response to rewards: an ERP study.

To study if the neural system responds the same or differently to the same rewards in different social competition conditions, event-related brain potentials (ERPs) were recorded as 18 participants engaged in two kinds of decision tasks. In the auction condition (the competition condition), participants were instructed to bid against their competitors and then informed the outcome (failing and gaining no money, or winning and gaining a certain amount of money); in the lottery condition (the neutral condition), subjects were asked to play a lottery against the computer and then informed the outcome (the same as in auction). Scalp ERPs revealed that, in the outcome phase, failing (rewards=0) in auction induced a larger late positive complex response (in the time window of 700-1200ms) than that in lottery; while winning (rewards>0) in lottery induced a larger late positive complex response (in the time window of 500-800ms) than that in auction. The present data suggest, when evaluating rewards, our neural systems care about not only rewards themselves, but also how the rewards have been obtained.