Pathway Metabolites Research Articles

Targeted and untargeted urinary metabolomics of alkaptonuria patients using ultra high-performance liquid chromatography-tandem mass spectrometry.

Alkaptonuria (AKU) is a rare autosomal-recessive disease which is characterized through black urine and ochronosis. It is caused by deficiency of the enzyme Homogentisate 1,2-dioxygenase in the Phenylalanine/Tyrosine degradation pathway which leads to the accumulation of Homogentisic acid (HGA). Urine was provided by AKU patients and healthy controls. Several different methods were developed in this study each with a specific goal.Firstly, a simple and inexpensive RP-UHPLC-UV method for routine monitoring of HGA as a key metabolite employing a Phenylhexyl stationary phase chemistry. Validation was performed in accordance to FDA guidelines and method selectivity was further evaluated via on-line high-resolution sampling 2D-LC-QToF-MS, coupling the Phenylhexyl phase in the first dimension with a C18 phase in the second dimension. Secondly, a targeted and accurate RP-UHPLC-MRM-QTRAP assay, providing quantitative analysis of the relevant pathway metabolites based on a Phenylhexyl stationary phase, and lastly an untargeted HILIC-UHPLC-QToF-MS/MS method with SWATH (sequential window acquisition of all theoretical mass spectra) acquisition employing a sulfobetaine-type HILIC-Z superficially porous particle column, with the aim of uncovering more details about the metabolic profile of this genetic disorder. By untargeted analysis 204 metabolites could be detected and annotated in positive and negative ESI mode in total. Two separate LC methods were employed, differing in their conditions depending on the ionization mode (20 mM ammonium formate as buffer additive adjusted to a pH = 3.5 with formic acid in ESI+ mode and 20 mM ammonium acetate adjusted to a pH = 7.5 with acetic acid in ESI- mode). By effectively combining the aforementioned methods, a comprehensive workflow was developed, allowing the effective analysis of both patient and control urine samples.

Oxidative and Excitatory Neurotoxic Stresses in CRISPR/Cas9-Induced Kynurenine Aminotransferase Knockout Mice: A Novel Model for Despair-Based Depression and Post-Traumatic Stress Disorder.

Memory and emotion are especially vulnerable to psychiatric disorders such as post-traumatic stress disorder (PTSD), which is linked to disruptions in serotonin (5-HT) metabolism. Over 90% of the 5-HT precursor tryptophan (Trp) is metabolized via the Trp-kynurenine (KYN) metabolic pathway, which generates a variety of bioactive molecules. Dysregulation of KYN metabolism, particularly low levels of kynurenic acid (KYNA), appears to be linked to neuropsychiatric disorders. The majority of KYNA is produced by the aadat (kat2) gene-encoded mitochondrial kynurenine aminotransferase (KAT) isotype 2. Little is known about the consequences of deleting the KYN enzyme gene. In CRISPR/Cas9-induced aadat knockout (kat2-/-) mice, we examined the effects on emotion, memory, motor function, Trp and its metabolite levels, enzyme activities in the plasma and urine of 8-week-old males compared to wild-type mice. Transgenic mice showed more depressive-like behaviors in the forced swim test, but not in the tail suspension, anxiety, or memory tests. They also had fewer center field and corner entries, shorter walking distances, and fewer jumping counts in the open field test. Plasma metabolite levels are generally consistent with those of urine: antioxidant KYNs, 5-hydroxyindoleacetic acid, and indole-3-acetic acid levels were lower; enzyme activities in KATs, kynureninase, and monoamine oxidase/aldehyde dehydrogenase were lower, but kynurenine 3-monooxygenase was higher; and oxidative stress and excitotoxicity indices were higher. Transgenic mice displayed depression-like behavior in a learned helplessness model, emotional indifference, and motor deficits, coupled with a decrease in KYNA, a shift of Trp metabolism toward the KYN-3-hydroxykynurenine pathway, and a partial decrease in the gut microbial Trp-indole pathway metabolite. This is the first evidence that deleting the aadat gene induces depression-like behaviors uniquely linked to experiences of despair, which appear to be associated with excitatory neurotoxic and oxidative stresses. This may lead to the development of a double-hit preclinical model in despair-based depression, a better understanding of these complex conditions, and more effective therapeutic strategies by elucidating the relationship between Trp metabolism and PTSD pathogenesis.

ApWD40a, a Member of the WD40-Repeat Protein Family, Is Crucial for Fungal Development, Toxin Synthesis, and Pathogenicity in the Ginseng Alternaria Leaf Blight Fungus Alternaria panax.

Alternaria panax, the primary pathogen that causes ginseng Alternaria leaf blight disease, can lead to a 20-30% reduction in ginseng yield. WD40 repeat-containing proteins are evolutionarily conserved proteins with diverse functions between different organisms. In this study, we characterized the roles of a WD40 repeat-containing protein in A. panax. The deletion of ApWD40a impaired the mycelial growth, reduced the sporulation, and significantly decreased the efficiency in utilizing various carbon sources. The ΔApwd40a mutant showed increased sensitivity to osmotic stress and metal ion stress induced by sorbitol, NaCl, and KCl, but decreased the sensitivity to a cell wall stress factor (SDS) and oxidative stress factors (paraquat and H2O2). Pathogenicity assays performed on detached ginseng leaves and roots revealed that the disruption of ApWD40a significantly decreased the fungal virulence through attenuating melanin and mycotoxin production by A. panax. A comparative transcriptome analysis revealed that ApWD40a was involved in many metabolic and biosynthetic processes, including amino acid metabolism, carbon metabolism, sulfate metabolic pathways, and secondary metabolite pathways. In particular, a significantly upregulated gene that encoded a sulfate permease 2 protein in ΔApwd40a, named ApSulP2, was deleted in the wild-type strain of A. panax. The deletion of ApSulP2 resulted in reduced biomass under sulfate-free conditions, demonstrating that the sulfate transport was impaired. Taken together, our findings highlight that ApWD40a played crucial roles in different biological processes and the pathogenicity of A. panax through modulating the expressions of genes involved in various primary and secondary metabolic processes.

Constitutive loss of kynurenine-3-monooxygenase changes circulating kynurenine metabolites without affecting systemic energy metabolism.

Kynurenic acid (KYNA) and quinolinic acid (QUIN) are metabolites of the kynurenine pathway of tryptophan degradation with opposing biological activities in the central nervous system. In the periphery, KYNA is known to positively affect metabolic health, whereas the effects of QUIN remain less explored. Interestingly, metabolic stressors, including exercise and obesity, differentially change the balance between circulating KYNA and QUIN. Here, we hypothesized that chronically elevated levels of circulating KYNA and reduced levels of QUIN would manifest as differences in whole-body energy metabolism. To test this, we used a mouse model lacking the enzyme kynurenine 3-monooxygenase (KMO), thus shunting kynurenine away from QUIN synthesis and towards KYNA production. KMO-deficient and wild-type littermate male and female mice were evaluated under chow and high-fat diets. Comprehensive kynurenine pathway metabolite profiling in plasma showed that the loss of KMO elicits robust changes in circulating levels of kynurenine metabolites. This included a 45-fold increase in kynurenine, a 26-fold increase in KYNA, and a 99% decrease in QUIN levels, depending on the diet. However, despite these changes, loss of KMO did not significantly impact whole-body energy metabolism or change the transcriptomic profile of subcutaneous adipose tissue on either diet. With KMO inhibitors being considered as therapeutic candidates for various disorders, this work shows that chronic systemic KMO inhibition does not have widespread metabolic effects. Our data also indicates that the beneficial effects of KYNA on metabolism may depend on its acute, intermittent elevation in circulation, akin to transient exercise-induced signals that mediate improved metabolic health.

Mid-life anti-inflammatory metabolites are inversely associated with long-term cardiovascular disease events.

Preclinical data have shown that low levels of metabolites with anti-inflammatory properties may impact metabolic disease processes. However, the association between mid-life levels of such metabolites and long-term ASCVD risk is not known. We characterised the plasma metabolomic profile (1228 metabolites) of 1852 participants (58.1±7.5 years old, 69.6% female, 43.6% self-identified as Black) enrolled in the Heart Strategies Concentrating on Risk Evaluation (Heart SCORE) study. Logistic regression was used to assess the impact of metabolite levels on ASCVD risk (nonfatal MI, revascularisation, and cardiac mortality). We additionally explored the effect of genetic variants neighbouring ASCVD-related genes on the levels of metabolites predictive of ASCVD events. The Atherosclerosis Risk in Communities (ARIC) study (n=4790; 75.5±5.1 years old, 57.4% female, 19.5% self-identified as Black) was used as an independent validation cohort. In fully adjusted models, alpha-ketobutyrate [AKB] (OR 0.62 [95% CI, 0.49-0.80]; p<0.001), and 1-palmitoyl-2-linoleoyl-GPI [OR, 0.62, 95% CI, 0.47-0.83; p<0.001], two metabolites in amino acid and phosphatidylinositol lipid pathways, respectively, showed a significant protective association with incident ASCVD risk in both Heart SCORE and ARIC cohorts. Three plasmalogens and a bilirubin derivative, whose levels were regulated by genetic variants neighbouring FADS1 and UGT1A1, respectively, exhibited a significant protective association with ASCVD risk in the Heart SCORE only. Higher mid-life levels of AKB and 1-palmitoyl-2-linoleoyl-GPI metabolites may be associated with lower risk late-life ASCVD events. Further research can determine the causality and therapeutic potential of these metabolites in ASCVD. This study was funded by the Pennsylvania Department of Health (ME-02-384). The department specifically disclaims responsibility for any analyses, interpretations, or conclusions. Additional funding was provided by National Institutes of Health (NIH) grant R01HL089292 and UL1 TR001857 (Steven Reis). Further, NIH funded R01HL141824 and R01HL168683 were used for the ARIC study validation (Bing Yu).

A 2OGD multi-gene cluster encompasses functional and tissue specificity that direct furanocoumarin and pyranocoumarin biosynthesis in citrus.

Furanocoumarins (FCs) are plant defence compounds derived from the phenylpropanoid pathway via the coumarin umbelliferone that harbour some therapeutic benefits yet are the underlying cause of 'grapefruit-drug interactions' in humans. Most of the pathway genes have not been identified in citrus. We employed a genetic/Omics approach on citrus ancestral species and F1 populations of mandarin × grapefruit and mandarin × pummelo. Enzyme specificity was characterized by Invivo 2-oxoglutarate-dependent dioxygenase family (2OGD) activity assays. We identified a 2OGD multi-gene cluster involved in coumarin/FC/pyranocoumarin biosynthesis; Species lacking FCs in leaves/fruit were homozygous for a 655-base solo-LTR frame-disrupting insertion within one dual specificity C2'H/F6'H encoding 2OGD gene, demonstrating that integrity of this gene is fully correlated with the capacity to biosynthesize metabolites of the extended FC pathway in leaves/fruit. A second 2OGD is the prominent gene expressed in citrus roots, which contain a unique pattern of extended FC pathway metabolites, including the predominant pyranocoumarins. A third 2OGD gene encodes a single activity F6'H, which appears to be induced at the transcript level by citrus pathogens. The results provide insights into the genetic basis underlying the difference between citrus fruit FC producers (grapefruit and pummelo) and nonproducers (mandarin and orange) and provide a gene target to breed for FC-free varieties by marker-assisted breeding or genome editing.

Associations of urinary caffeine metabolites with sex hormones: comparison of three statistical models.

The association between urinary caffeine and caffeine metabolites with sex hormones remains unclear. This study used three statistical models to explore the associations between urinary caffeine and its metabolites and sex hormones among adults. We selected the participants aged ≥18 years in the National Health and Nutrition Examination Survey (NHANES) data 2013-2014 as our study subjects. We performed principal components analysis (PCA) to investigate the underlying correlation structure of urinary caffeine and its metabolites. Then we used these principal components (PCs) as independent variables to conduct multiple linear regression analysis to explore the associations between caffeine metabolites and sex hormones (E2, TT, SHBG). We also fitted weighted quantile sum (WQS) regression, and Bayesian kernel machine regression (BKMR) methods to further assess these relationships. In the PCA-multivariable linear regression, PC2 negatively correlates with E2: β = -0.01, p-value = 0.049 (male population). In the WQS regression model, the WQS indices were associated with SHBG and TT both in male (SHBG: WQS index = -0.11, p < 0.001; TT: WQS index = -0.10, p < 0.001) and female (SHBG: WQS index = -0.10, p < 0.001; TT: WQS index = -0.04, p < 0.001) groups. Besides, the WQS index was significantly associated with E2 in females (p < 0.05). In the BKMR model, despite no significant difference in the overall association between caffeine metabolites and the sex hormones (E2, TT, SHBG), there was nonetheless a declining trend in the male population E2 group, in the male and female population SHBG groups also observed a downward trend. When considering the results of these three models, the whole-body burden of caffeine metabolites, especially the caffeine metabolites in the PC2 metabolic pathway was significantly negatively associated with E2 in males. Considering the advantages and disadvantages of the three statistical models, we recommend applying diverse statistical methods and interpreting their results together.

Mechanisms and applications of bacterial luciferase and its auxiliary enzymes.

Bacterial luciferase (LuxAB) catalyzes the conversion of reduced flavin mononucleotide (FMNH⁻), oxygen, and a long-chain aldehyde to oxidized FMN, the corresponding acid and water with concomitant light emission. This bioluminescence reaction requires the reaction of a flavin reductase such as LuxG (in vivo partner of LuxAB) to supply FMNH⁻ for the LuxAB reaction. LuxAB is a well-known self-sufficient luciferase system because both aldehyde and FMNH⁻ substrates can be produced by the associated enzymes encoded by the genes in the lux operon, allowing the system to be auto-luminous. This makes it useful for in vivo applications. Structural and functional studies have long been performed in efforts to gain a better understanding of the LuxAB reaction. Recently, continued exploration of the LuxAB reaction have elucidated the mechanisms of C4a-hydroperoxyflavin formation and identified key catalytic residues such as His44 that facilitates the generation of flavin intermediates important for light generation. Advancements in protein engineering and synthetic biology have improved the bioluminescence properties of LuxAB. Various applications of LuxAB for bioimaging, bioreporters, biosensing in metabolic engineering and real-time monitoring of aldehyde metabolites in biofuel production pathways have been developed during the last decade. Challenging issues such as achieving red-shifted emissions, optimizing the signal intensity and identifying mechanisms related to the generation of light-emitting species remain to be explored. Nevertheless, LuxAB continues to be a promising tool for diverse biotechnological and biomedical applications.

Targeted and non-targeted metabolic analysis of chlorpromazine in grass carp as well as the in-silico and metabolomics toxicity assessment.

Chlorpromazine (CPZ) is an abused sedative that is extensively metabolized in organisms. However, the metabolic pathway of CPZ in aquatic organisms is still unclear. In this study, CPZ metabolites was analyzed in grass carp exposed to CPZ in the raising water using ultrahigh-performance liquid chromatography coupled with quadrupole Orbitrap mass spectrometry (UHPLC-Q-Orbitrap MS). Thirteen CPZ metabolites were identified, including 11 previously reported and 2 newly identified metabolites (M9 and M13), and 5 known metabolites were confirmed using authentic standards. The molecular structures and transformation pathways of CPZ metabolites were putatively deduced, which mainly included oxygenation, demethylation, dechlorination and carboxylation reactions. Quantitative analysis of CPZ and its metabolites were also performed, and CPZ sulfoxide had a higher content as an important characteristic metabolite. In addition, in-silico toxicity prediction reminded that some metabolites possess ecotoxicity and developmental toxicities similar to, or even higher, than CPZ. Moreover, metabolomics results indicated that CPZ exposure could cause metabolic disorder in the endogenous metabolome of grass carp.

Comparative Metabolomics in Liver Reveals Eggshell Translucency Formation Using LC-MS Analysis

Eggshell translucency is the common problem on eggshell that become more severe with age. They are important because it influence consumer preferences and the economic value of eggs. The reason for the eggshell translucency is currently believed to be the eggshell membrane (ESM). In this study, we screened translucency eggs and normal eggs and used metabolomics to study liver metabolism in different eggshell translucency and discuss important liver lipids and phosphatidyl metabolites. Liver samples were taken for Liquid Chromatograph Mass Spectrometer (LC-MS) during the formation of eggshell membranes in hens, that is, when eggs form eggshell membranes in the oviduct isthmus. The results showed that we identified two essential metabolic pathways through differential metabolite pathway analysis, which were glycine, serine, and threonine metabolism related to amino acids metabolism and the PPAR metabolic pathway related to lipid metabolism. Furthermore, this study helps us understand the process of translucency egg production in poultry. This laid the foundation for in-depth research on eggshell translucency. These results may and provide support for future breeding.

Ventricular septal defects in cats – a retrospective study

Significant deterioration in semen quality is associated with increased male infertility. Alterations in the sperm proteome resulting from infertility are not well understood. The aim of this study was to examine the connection between the proteome and the quality of spermatozoa in fertile and infertile dogs. The research was conducted on 11 male dogs, including 3 German shepherds, 3 Turkish Kangal, 2 Cane Corso, and 3 Golden Retrievers, aged 4 and 6 years (4.81 ± 0.75). Four of the 11 dogs constituted the infertile study group because of conception failure observed in at least 3 (3-5) matings with different fertile females within 1 year, and seven dogs from different owners constituted the fertile control group. Semen was manually manipulated and collected from all dogs in the presence of a female dog in heat. Ejaculates were used for spermatological examination and protein analysis. Computer-assisted semen analysis (CASA) was used to evaluate semen samples. In spermatological parameters, motility, kinematic parameters, morphology and plasma membrane integrity were evaluated. Gel electrophoresis (SDS-PAGE) was used to analyze proteins in sperm. The semen quality of infertile dogs was significantly lower. Motility, progressive motility, viability and plasma membrane integrity were statistically significantly lower in the infertile group than in the fertile group (P &lt; 0.05). In morphological parameters, acrosome, head, middle part, tail and total defects were found to be statistically significantly higher in sterile dogs than in fertile dogs (P &lt; 0.05). In the study, a total of 8 protein bands with molecular weights (MW) ranging from 11.0 to 245.0 kDa were identified in the protein analysis of semen. Variations in the sperm proteome composition were shown to be dependent on fertility. These proteins were associated with important metabolite pathways. Additionally, correlations of these bands with various spermatological parameters were revealed. The result of the study suggests that seminal plasma proteins play a role in semen quality and may be potential fertility biomarkers.

Characterization of Fertile and Infertile Dog Seminal Plasma Proteins and Their Correlation with Semen Quality: Preliminary Study

Significant deterioration in semen quality is associated with increased male infertility. Alterations in the sperm proteome resulting from infertility are not well understood. The aim of this study was to examine the connection between the proteome and the quality of spermatozoa in fertile and infertile dogs. The research was conducted on 11 male dogs, including 3 German shepherds, 3 Turkish Kangal, 2 Cane Corso, and 3 Golden Retrievers, aged 4 and 6 years (4.81 ± 0.75). Four of the 11 dogs constituted the infertile study group because of conception failure observed in at least 3 (3-5) matings with different fertile females within 1 year, and seven dogs from different owners constituted the fertile control group. Semen was manually manipulated and collected from all dogs in the presence of a female dog in heat. Ejaculates were used for spermatological examination and protein analysis. Computer-assisted semen analysis (CASA) was used to evaluate semen samples. In spermatological parameters, motility, kinematic parameters, morphology and plasma membrane integrity were evaluated. Gel electrophoresis (SDS-PAGE) was used to analyze proteins in sperm. The semen quality of infertile dogs was significantly lower. Motility, progressive motility, viability and plasma membrane integrity were statistically significantly lower in the infertile group than in the fertile group (P &lt; 0.05). In morphological parameters, acrosome, head, middle part, tail and total defects were found to be statistically significantly higher in sterile dogs than in fertile dogs (P &lt; 0.05). In the study, a total of 8 protein bands with molecular weights (MW) ranging from 11.0 to 245.0 kDa were identified in the protein analysis of semen. Variations in the sperm proteome composition were shown to be dependent on fertility. These proteins were associated with important metabolite pathways. Additionally, correlations of these bands with various spermatological parameters were revealed. The result of the study suggests that seminal plasma proteins play a role in semen quality and may be potential fertility biomarkers.

A NEW ROLE FOR KYNURENINE METABOLISM IN THE AGE-ASSOCIATED DECLINE OF THE BACTERIAL IMMUNE RESPONSE

Abstract Tryptophan metabolism through the kynurenine pathway becomes dysregulated during normal aging and is implicated in age-associated disease, including chronic inflammation, atherosclerosis, neurodegeneration, and cancer. Kynurenine pathway enzymes and metabolites influence a range of molecular processes critical to healthy aging, including regulation of inflammatory and immune responses. Kynurenine metabolism is active in immune cells and activated in response to proinflammatory cytokine signaling. We previously determined that elevating physiological levels of the kynurenine pathway metabolite 3-hydroxyanthranilic acid (3HAA) via either direct supplementation or inhibition of the enzyme that degrades 3HAA, 3HAA dioxygenase (HAAO), extends lifespan and delays age-associated health decline in both Caenorhabditis elegans and mice. Published work suggests that 3HAA is anti-inflammatory in mammals, for example by reducing the ratio of activated to regulatory T cells and inhibiting pathological activation of macrophages. In recent work, we find that elevating physiological 3HAA can beneficially enhance the immune response of C. elegans to bacterial pathogens during aging. 3HAA is sufficient to kill bacterial and inhibit bacterial growth in culture. When HAAO is inhibited in C. elegans, 3HAA accumulates in lysosome related organelles (LROs) in the intestinal cells, the same subcellular compartment that contains engulfed bacteria. LROs are a cellular repository for both iron and zinc, and we further find that iron chelation or zinc supplementation dramatically enhances the bactericidal properties of 3HAA. Here we present a mechanistic model in which age-dependent accumulation of 3HAA in intestinal LROs combines with zinc to enhance bacterial resistance with age in C. elegans.

Dietary Influences on Gut Microbiota and Their Role in Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD).

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a major contributor to liver-related morbidity, cardiovascular disease, and metabolic complications. Lifestyle interventions, including diet and exercise, are first line in treating MASLD. Dietary approaches such as the low-glycemic-index Mediterranean diet, the ketogenic diet, intermittent fasting, and high fiber diets have demonstrated potential in addressing the metabolic dysfunction underlying this condition. The development and progression of MASLD are closely associated with taxonomic shifts in gut microbial communities, a relationship well-documented in the literature. Given the importance of diet as a primary treatment for MASLD, it is important to understand how gut microbiota and their metabolic byproducts mediate favorable outcomes induced by healthy dietary patterns. Conversely, microbiota changes conferred by unhealthy dietary patterns such as the Western diet may induce dysbiosis and influence steatotic liver disease through promoting hepatic inflammation, up-regulating lipogenesis, dysregulating bile acid metabolism, increasing insulin resistance, and causing oxidative damage in hepatocytes. Although emerging evidence has identified links between diet, microbiota, and development of MASLD, significant gaps remain in understanding specific microbial roles, metabolite pathways, host interactions, and causal relationships. Therefore, this review aims to provide mechanistic insights into the role of microbiota-mediated processes through the analysis of both healthy and unhealthy dietary patterns and their contribution to MASLD pathophysiology. By better elucidating the interplay between dietary nutrients, microbiota-mediated processes, and the onset and progression of steatotic liver disease, this work aims to identify new opportunities for targeted dietary interventions to treat MASLD efficiently.

A positively charged carbon dot complex improves the bioactivity of Isaria fumosorosea against Plutella xylostella (Linnaeus)

BackgroundNanomaterials compounded with entomopathogenic fungi have been proven to be effective in insect pest management. This study reports the synthesis and bioactivity of Isaria fumosorosea-based nanoparticles by compounding the fungus with differentially (positively or negatively) charged carbon dots. Initially, negatively charged carbon dots (N-CDs) and carbon dots (P-CDs) were synthesized via a one-step hydrothermal method. The N-CDs and P-CDs were then individually compounded with I. fumosorosea to develop N-CD@I. fumosorosea and P-CD@I. fumosorosea nanocomposites.ResultsCharacterization of the nanoparticles revealed that positively or negatively charged carbon dots were attached to I. fumosorosea by electrostatic bonding. Finally, the virulence of both types of nanoparticles was observed in Plutella xylostella. The bioassay results indicated that the highest P. xylostella mortality (92.7 ± 2.04%) was associated with the P-CD@I. fumosorosea treatment. The results of nontargeted metabolomic analysis revealed that different treatments affected the metabolism of P. xylostella by interfering with the riboflavin metabolism pathway by downregulating the expression of two main metabolites of the riboflavin metabolism pathway (2-5-diamino-6-pyrimidin-4-one and 7-hydroxy-6-methyl-8-ribiotin), although the extent of the expression downregulation varied among the different treatments.ConclusionOverall, this work provides insight into the mechanism by which positively charged nanomaterials improve the virulence of entomopathogenic fungi. This work provides a new direction for the design and application of nanomaterials for insect pest management.Graphical

Integrated proteomics and metabolomics network analysis across different delivery modes in human pregnancy: a pilot study

BackgroundDelivery mode has been linked to child health, e.g., allergic disease. However, it remains unclear whether protein and metabolite differences across different delivery modes may underlie child development.MethodsA cohort comprising 16 spontaneous onset vaginal delivery (VD), 16 prelabor cesarean delivery on maternal request (CS), and 8 intrapartum cesarean section (Intra_CS) women were analyzed using label-free proteomic and untargeted metabolomics assays on amniotic fluid and cord blood samples, respectively. We used weighted gene co-expression network analyses (WGCNA) to identify modules of highly correlated proteins or metabolites that associated with delivery modes and related clinical traits. KEGG enrichment analyses were performed to investigate the biological function of the identified modules. Integrative multiomics analysis was employed to examine the biological interplay between proteomic and metabolic interactions.ResultsCompared to the CS group, the proteomic and metabolomic profiles were similar between the Intra_CS and VD groups in our study. We did not identify any enriched protein or metabolite pathways related to immune development that could influence the risk of allergic diseases in offspring across different delivery modes. However, we identified seven protein modules correlated with the duration from the rupture of the membranes to full dilation of the cervix, with the actin cytoskeleton module significantly enriched. A metabolic module in cord blood that correlated with VD was enriched in subclasses including C21 steroids, steroid sulfates, and oxysterols. Integrative analysis of proteomic and metabolomic data suggested pathways related to mode of delivery and duration of labor, encompassing the actin cytoskeleton, NADP metabolic process, nicotinate, and nicotinamide metabolism in amniotic fluid, and the steroid hormone biosynthesis pathway in cord blood.ConclusionsDifferences in steroid hormones and the actin cytoskeleton pathway according to proteomics and metabolomics in amniotic fluid and cord blood were more indicative of the labor process. These findings could guide future studies on delivery-associated biochemical pathways.

Whole Genome Identification and Integrated Analysis of Long Non-Coding RNAs Responding ABA-Mediated Drought Stress in Panax ginseng C.A. Meyer.

Panax ginseng C.A. Meyer is a perennial herb that is used worldwide for a number of medical purposes. Long non-coding RNAs (lncRNAs) play a crucial role in diverse biological processes but still remain poorly understood in ginseng, which has limited the application of molecular breeding in this plant. In this study, we identified 17,478 lncRNAs and 3106 novel mRNAs from ginseng by high-throughput illumine sequencing. 50 and 257 differentially expressed genes (DEGs) and DE lncRNAs (DELs) were detected under drought + ABA vs. drought conditions, respectively. The DEGs and DELs target genes main enrichment is focused on the "biosynthesis of secondary metabolites", "starch and sucrose metabolism", and "carbon metabolism" pathways under drought + ABA vs. drought conditions according to KEGG pathway enrichment analysis, suggesting that these secondary metabolites biosynthesis pathways might be crucial for ABA-mediated drought stress response in ginseng. Together, we identified drought stress response lncRNAs in ginseng for the first time and found that the target genes of these lncRNAs mainly regulate the biosynthesis of secondary metabolites pathway to response to drought stress. These findings also open up a new visual for molecular breeding in ginseng.

Microbial diversity and metabolomics analysis of colon contents exposed to cadmium and polystyrene microplastics.

Cadmium and microplastics, common pollutants, can accumulate in the body, impacting the intestinal barrier and harming livestock breeding. In order to explore the damage mechanism of cadmium and cadmium combined microplastic on the colon of mice, 60 mice were divided into three groups: The control group (0.2 mL of saline), cadmium group (Cd group, 0.2 mL of 4.8 mg/kg/d CdCl2) and mixed group (Mix group, 0.2 mL of mixed solution containing 4.8 mg/kg/d CdCl2 and 10.0 mg/d MPs) were fed for 42 d. The changes of colon histopathology were observed, and the changes of microbial diversity and metabolomics of colon contents were analyzed. Pathological sections of the colon showed abnormal mucosal hyperemia with mixed exposure compared to cadmium exposure. Microbial diversity analysis showed increased abundances of Enterococcus, Adlercreutzia, and Bifidobacterium in the Cd and Mix groups, with Dubosiella being the most significantly increased. Metabolomic analysis indicated significant differences in nucleotide and purine metabolism between the Cd and control groups, and in linoleic acid and bile acid metabolism between the Mix and control groups. The ABC transporter metabolites increased with Cd exposure, while the PPAR pathway metabolites were enriched with MPs exposure. Correlation analysis highlighted several key findings: Pasteurella exhibited a notably negative association with pantothenate. Conversely, Enterococcus demonstrated a significant positive link with palmitoylcarnitine. Additionally, both Adlercreutzia and norank_f_Eggerthellaceae showed a positive correlation with azelaic acid. These findings suggest that Cd and MPs disrupt intestinal microbiota and metabolic pathways, providing insights into potential treatments for such exposures.

Green Radish Polysaccharides Ameliorate Hyperlipidemia in High-Fat-Diet-Induced Mice via Short-Chain Fatty Acids Production and Gut Microbiota Regulation.

The objective of this study was to examine the hypolipidemic effect and potential mechanism of action of green radish polysaccharide (GRP) in hyperlipidemic mice. We found that in mice fed a high-fat diet, supplementing with GRP reduced body weight and liver index, significantly improved serum lipid levels and markers of liver damage, and mitigated oxidative stress and inflammation. Mechanistically, in these hyperlipidemic mice, the size of fat cells was reduced by GRP, and the abnormal accumulation of lipid droplets was reduced. We also found that GRP regulates the composition of the intestinal microbiota, including the ratio of Firmicutes to Mycobacteria F/B and the levels of Blautia spp., which have been shown to alleviate liver damage and treat hyperlipidemia. Metabolite pathway analysis using the Kyoto Encyclopedia of Genes and Genomes identified the glycolysis/glycolytic metabolism and propionate metabolism pathways as potential targets for GRP in the amelioration of hyperlipidemia.

The Association of Fructose Metabolism With Anesthesia/Surgery-Induced Lactate Production.

In elderly individuals, excessive lactate levels in the brain may be associated with the development of cognitive impairment after surgery, including delayed neurocognitive recovery (dNCR). Since the origin of this increased lactate is unknown, here we assessed associations between metabolic pathways and postoperative dNCR. This study included 43 patients (≥60 years old) who had surgery under general anesthesia. We also used a mouse model in which 20-month-old mice were exposed to sevoflurane to induce postoperative dNCR, while control mice were exposed to 40% oxygen. Mice in the control group and anesthesia/surgery group were injected with fructose or glucose intracerebroventricularly, or fructose metabolism inhibitor intraperitoneally. Barnes maze test and Y maze were used to measure cognitive function in mice. Metabolomics was used to measure metabolites in the serum of patients and the brains of mice after anesthesia/surgery. Isotope labeling and metabolic flux were used to analyze flow and distribution of specific metabolites in metabolic pathways. Among 43 patients, 17 developed dNCR. Metabolomics showed significantly decreased postoperative serum fructose 1-phosphate levels in dNCR compared to nondNCR patients (mean difference [×104] = -0.164 ± 0.070; P = .024). Similar results were found in the brains of mice (mean difference = -1.669 ± 0.555; *P = .014). Isotope labeling and metabolic flux experiments in mice showed fructose but not glucose entered glycolysis, increasing lactate levels in the brain after anesthesia/surgery (P < .05). Administration of intraperitoneal fructose inhibitors to mice effectively inhibited increased lactate levels in the brain (mean difference =96.0 ± 4.36, P = .0237) and cognitive dysfunction after anesthesia/surgery (mean difference =69.0 ± 3.94, P = .0237). In a small subsample, we also found anesthesia/surgery increased interleukin-6 (IL-6) levels in the brains of mice (mean difference =88.3 ± 3.44, P = .0237) and that IL-6 may function upstream in fructose activation. These results suggest that anesthesia/surgery activates fructose metabolism, producing excessive lactate in the brain that is associated with postoperative cognitive impairment. Fructose metabolism is thus a potential therapeutic target for dNCR.