PubMed

Redox Biol. 2023 Sep 6;67:102875. doi: 10.1016/j.redox.2023.102875. Online ahead of print.ABSTRACTHigher consumption of broccoli (Brassica oleracea var. italica) is associated with a reduced risk of cardiometabolic diseases, neurological disorders, diabetes, and cancer. Broccoli is rich in various phytochemicals, including glucosinolates, and isothiocyanates. Moreover, it has recently reported the endogenous production of polysulfides, such as cysteine hydropersulfide (CysS2H) and glutathione hydropersulfide (GS2H), in mammals including humans, and that these bioactive substances function as potent antioxidants and important regulators of redox signaling in vivo. However, few studies have focused on the endogenous polysulfide content of broccoli and the impact of germination on the polysulfide content and composition in broccoli. In this study, we investigated the alternations in polysulfide biosynthesis in broccoli during germination by performing untargeted polysulfide omics analysis and quantitative targeted polysulfide metabolomics through liquid chromatography-electrospray ionization-tandem mass spectrometry. We also performed 2,2-diphenyl-1-picrylhydrazyl radical-scavenging assay to determine the antioxidant properties of the polysulfides. The results revealed that the total polysulfide content of broccoli sprouts significantly increased during germination and growth; CysS2H and cysteine hydrotrisulfide were the predominant organic polysulfide metabolites. Furthermore, we determined that novel sulforaphane (SFN) derivatives conjugated with CysS2H and GS2H were endogenously produced in the broccoli sprouts, and the novel SFN conjugated with CysS2H exhibited a greater radical scavenging capacity than SFN and cysteine. These results suggest that the abundance of polysulfides in broccoli sprouts contribute to their health-promoting properties. Our findings have important biological implications for the development of novel pharmacological targets for the health-promoting effects of broccoli sprouts in humans.PMID:37699321 | DOI:10.1016/j.redox.2023.102875

Aquat Toxicol. 2023 Sep 7:106688. doi: 10.1016/j.aquatox.2023.106688. Online ahead of print.ABSTRACTDue to clinical treatment and illegal use, psychoactive substances have been widely detected in the aquatic environment. In this study, we investigated the effects of the benzodiazepine drug flunitrazepam (FLZ) and its metabolite 7-aminoflunitrazepam (7-FLZ) on the gut-liver axis of zebrafish. Zebrafish were exposed to two concentrations of FLZ and 7-FLZ (0.05 and 1 μg/L) for 30 days. Results showed that both FLZ and 7-FLZ exposure altered the relative abundance of Proteobacteria at the phylum level, with significant differences observed at the genus level for pathogenic bacteria such as Paracoccus, Shewanella, and Aeromonas. Metabolomics results showed both exposures significantly interfered with nucleotide and amino acid metabolism. The imbalance of gut microbiota and metabolic disorder increased the level of malondialdehyde, which in turn heightened the permeability of the gut mucosal barrier. FLZ and 7-FLZ induced oxidative stress in the liver via the gut-liver axis, leading to decreased levels of glucose, total cholesterol, and triglyceride, as well as the down-regulation of glycolipid metabolism-related genes (PPARα, PPARγ, FABP2, Fabp11, PFKFB3, and LDHA). Metabolomics results revealed that FLZ and 7-FLZ significantly affected the biosynthesis of amino acids and arginine, and other metabolic pathways such as nucleotide, nicotinate and nicotinamide, and purine in the liver. Our results unveiled the mechanisms behind the toxicological effects of psychoactive substances on the gut-liver axis, providing valuable data for ecological and environmental risk assessments.PMID:37699776 | DOI:10.1016/j.aquatox.2023.106688

JCI Insight. 2023 Sep 12:e162100. doi: 10.1172/jci.insight.162100. Online ahead of print.ABSTRACTGermline de novo missense variants of the CACNA1D gene, encoding the pore-forming α1-subunit of Cav1.3 L-type Ca2+ channels (LTCCs), have been found in patients with neurodevelopmental and endocrine dysfunction, but their disease-causing potential is unproven. These variants alter channel gating enabling enhanced Cav1.3 activity, suggesting Cav1.3 inhibition as a potential therapeutic option. Here we provide direct proof of the disease-causing nature of such gating-modifying CACNA1D variants using mice (Cav1.3AG) containing the A749G variant reported de novo in a patient with autism and intellectual impairment. In heterozygous mutants native LTCC currents in adrenal chromaffin cells exhibited gating changes as predicted from heterologous expression. The A749G mutation induced aberrant excitability of dorsomedial striatum-projecting substantia nigra dopamine neurons and medium spiny neurons in the dorsal striatum. The phenotype observed in heterozygous mutants reproduced many of the abnormalities described within the human disease spectrum, including developmental delay, social deficit and pronounced hyperactivity without major changes in gross neuroanatomy. Despite an ~7-fold higher sensitivity of A749G-containing channels to the LTCC inhibitor isradipine, oral pretreatment over two days did not rescue the hyperlocomotion. Cav1.3AG mice confirm the pathogenicity of the A749G variant and point towards a pathogenetic role of altered signaling in the dopamine midbrain system.PMID:37698939 | DOI:10.1172/jci.insight.162100

JCI Insight. 2023 Sep 12:e162468. doi: 10.1172/jci.insight.162468. Online ahead of print.ABSTRACTSomatic gain-of-function mutations in the L-type calcium channel CaV1.3 (CACNA1D gene) cause adrenal aldosterone-producing adenomas and micronodules. De novo germline mutations are found in a syndrome of primary aldosteronism, seizures and neurologic abnormalities (PASNA) as well as in autism spectrum disease. Using CRISPR/Cas9, we here generated mice with a Cacna1d gain-of-function mutation found in both adenomas and PASNA syndrome (Cacna1dIle772Met/+). These mice show reduced body weight and increased mortality from weaning to approximately 100 days of age. Male mice do not breed, likely due to neuromotor impairment, and the offspring of female mice die perinatally, likely due to lack of maternal care. Mice generated by in vitro fertilization show elevated intracellular calcium in the aldosterone-producing zona glomerulosa, an elevated aldosterone:renin ratio and persistently elevated serum aldosterone on a high-salt diet as signs of primary aldosteronism. Anesthesia with ketamine and xylazine induces tonic-clonic seizures. Neurologic abnormalities include hyperlocomotion, impaired performance in the rotarod test, impaired nest building and slight changes in social behavior. Intracellular calcium in the zona glomerulosa, aldosterone levels and rotarod performance respond to treatment with the calcium channel blocker isradipine, with implications for the therapy of patients with aldosterone-producing lesions and with PASNA syndrome.PMID:37698934 | DOI:10.1172/jci.insight.162468

Nephrol Dial Transplant. 2023 Sep 12:gfad201. doi: 10.1093/ndt/gfad201. Online ahead of print.ABSTRACTBACKGROUND AND HYPOTHESIS: Hyperuricemia is prevalent in individuals with chronic kidney disease. Elevated serum uric acid concentrations have been considered an independent risk factor for the onset of chronic kidney disease. However, the relationship between serum uric acid concentrations and long-term health outcomes among patients with chronic kidney disease remains unclear.METHODS: We performed a prospective cohort study with nationally representative sample samples to investigate the relationship between serum uric acid concentrations and mortality risk among patients with chronic kidney disease. The weighted restricted cubic spline analyses combined with the multivariate-adjusted Cox proportional hazard models were used to examine a non-linear relationship multivariate-adjusted on all-cause, cardiovascular disease, and cancer mortality.RESULTS: The 6642 patients participating in National Health and Nutrition Examination Survey 1999-2018 were enrolled. During 656 885 person-months of follow-up time, 2619 all-cause deaths were recorded, including 1030 cardiovascular disease deaths and 458 cancer deaths. Our study presented J-shaped non-linear relationships between serum uric acid concentrations and all-cause and cardiovascular disease mortality with inflection points at 311.65 μmol/L and 392.34 μmol/L, respectively. When serum uric acid concentration was higher than those inflection points, every increase of 50 μmol/L serum uric acid was associated with 11.7% and 17.0% greater multivariable-adjusted hazard ratios of all-cause and cardiovascular disease mortality, respectively. In addition, a negative linear correlation with cancer mortality was detected.CONCLUSION: These findings suggested that maintaining appropriate serum uric acid concentrations may improve long-term health outcomes among chronic kidney disease patients. The corresponding inflection points of J-shaped non-linear relationships were 311.65 and 392.34 μmol/L for all-cause and cardiovascular disease mortality. Further clinical trials are required to investigate uric acid lowering targets.PMID:37698875 | DOI:10.1093/ndt/gfad201

Metabolomics. 2023 Sep 12;19(9):82. doi: 10.1007/s11306-023-02046-2.ABSTRACTINTRODUCTION: The objective of this study was to explore potential novel biomarkers for moderate to severe lower urinary tract symptoms (LUTS) using a metabolomics-based approach, and statistical methods with significant different features than previous reported.MATERIALS AND METHODS: The patients and the controls were selected to participate in the study according to inclusion/exclusion criteria (n = 82). We recorded the following variables: International prostatic symptom score (IPSS), prostate volume, comorbidities, PSA, height, weight, triglycerides, glycemia, HDL cholesterol, and blood pressure. The study of 41 plasma metabolites was done using the nuclear magnetic resonance spectroscopy technique. First, the correlations between the metabolites and the IPSS were done using Pearson. Second, significant biomarkers of LUTS from metabolites were further analysed using a multiple linear regression model. Finally, we validated the findings using partial least square regression (PLS).RESULTS: Small to moderate correlations were found between IPSS and methionine (-0.301), threonine (-0.320), lactic acid (0.294), pyruvic acid (0.207) and 2-aminobutyric-acid (0.229). The multiple linear regression model revealed that only threonine (p = 0.022) was significantly associated with IPSS, whereas methionine (p = 0.103), lactic acid (p = 0.093), pyruvic acid (p = 0.847) and 2-aminobutyric-acid (p = 0.244) lost their significance. However, all metabolites lost their significance in the PLS model.CONCLUSION: When using the robust PLS-regression method, none of the metabolites in our analysis had a significant association with lower urinary tract symptoms. This highlights the importance of using appropriate statistical methods when exploring new biomarkers in urology.PMID:37698748 | DOI:10.1007/s11306-023-02046-2

Stress Biol. 2023 Sep 12;3(1):39. doi: 10.1007/s44154-023-00118-w.ABSTRACTRice blast, caused by Magnaporthe oryzae, is one of the most destructive rice diseases. Developing blast-resistant rice cultivars represents the most economical and environmentally friend strategy for managing the disease. In our previous study, an isobaric tags for relative and absolute quantitation (iTRAQ)-based comparative protein quantification was carried out to investigate the resistance gene Piz-t gene-mediated resistance response to infection in two contrasting rice genotypes of the Piz-t transgenic Nipponbare line (NPB-Piz-t) and its wild-type Nipponbare (NPB). Here, from the comparisons of differentially expressed proteins (DEPs) of NPB-Piz-t to the avirulent isolate KJ201 (KJ201-Piz-t)and the virulent isolate RB22 (RB22-Piz-t) with mock-treated NPB-Piz-t (Mock-Piz-t), NPB to the virulent isolate KJ201(KJ201-NPB) and RB22 (RB22-NPB) with mock-treated NPB (Mock-NPB), 1, 1, and 6 common DEPs were, respectively, identified at 24, 48 and 72 h post-inoculation (hpi) in the susceptible comparisons of RB22-Pizt/Mock-Piz-t, KJ201-NPB/Mock-NPB, and RB22-NPB/Mock-NPB, involving in gi|54,290,836 and gi|59,800,021 were identified in the resistance comparison KJ201-Piz-t/Mock-Piz-t at 48 and 72 hpi respectively. Moreover, four genes of Os01g0138900 (gi|54,290,836), Os04g0659300 (gi|59,800,021), Os09g0315700 (gi|125,563,186) or Os04g0394200 (gi|21,740,743) were knocked out or overexpressed in NPB using gene over-expression and CRISPR/Cas9 technology, and results verified that the Os01g0138900 obviously affected the rice blast resistance. Further, expression and targeted metabolomics analysis illuminated the resistance response of cysteine-containing substances as gi|59,800,021 under blast infection. These results provide new targets for basal resistance gene identification and open avenues for developing novel rice blast resistant materials.PMID:37698658 | DOI:10.1007/s44154-023-00118-w

mSystems. 2023 Sep 12:e0068323. doi: 10.1128/msystems.00683-23. Online ahead of print.ABSTRACTOral diseases, such as periodontal disease and dental caries, have a high risk of recurrence and are considered to be related to dysbiosis of the oral microbiome and metabolome. It is, therefore, important to understand the state of the oral microbiome and metabolome after disease treatment to prevent recurrence. The current study sought to clarify whether oral dysbiosis improves the following remission of symptoms by dental treatment. To this end, the salivary microbiome and metabolome of healthy participants were compared to those with periodontal disease, dental caries, or both (concurrent). Saliva was collected as mouth-rinsed water and the microbiome was measured using 16S rRNA gene targeted sequencing and metabolome using capillary electrophoresis-time-of-flight mass spectrometry. Comparisons with healthy participants were performed before and after treatment, and several months after transition to self-care. Dental treatment significantly improved the oral health condition of each group; several months after treatment, oral health did not deteriorate. However, even after remission, the salivary microbiome of the two groups (oral disease and healthy) differed significantly. Additionally, after remission, significant differences, compared with healthy participants, remained in the relative abundances of disease-related bacteria and nitrate-reducing bacteria. After remission, significant differences were observed in the salivary metabolome in the healthy group in terms of threonate and pyrimidine metabolism-related component concentrations, which are assumed to reflect the high relative abundance of periodontal disease-related bacteria in the microbiome. Oral microbiome dysbiosis persisted even after dental treatment-induced disease remission with a sustained increased risk of disease when compared with healthy participants. IMPORTANCE We characterized the oral conditions, salivary microbiome, and metabolome after dental treatment by investigating the state after treatment completion and transition to self-care. Dental treatment improved oral health conditions, resulting in oral disease remission; however, the imbalanced state of the salivary microbiome continued even after remission. Although the results of this study are preliminary, owing to the small number of participants in each group when compared to larger cohort studies, they indicate that the risk of disease may remain higher than that of healthy participants, thereby demonstrating the importance of removing dental plaque containing disease-related bacteria using appropriate care even after treatment completion. We also identified bacterial species with relative abundances that differed from those of healthy participants even after remission of symptoms, which may indicate that the maturation of certain bacterial species must be controlled to improve the oral microbiome and reduce the risk of disease recurrence.PMID:37698410 | DOI:10.1128/msystems.00683-23

FASEB J. 2023 Oct;37(10):e23184. doi: 10.1096/fj.202300840RR.ABSTRACTExercise is a major beneficial contributor to muscle metabolism, and health benefits acquired by exercise are a result of molecular shifts occurring across multiple molecular layers (i.e., epigenome, transcriptome, and proteome). Identifying robust, across-molecular level targets associated with exercise response, at both group and individual levels, is paramount to develop health guidelines and targeted health interventions. Sixteen, apparently healthy, moderately trained (VO2 max = 51.0 ± 10.6 mL min-1 kg-1 ) males (age range = 18-45 years) from the Gene SMART (Skeletal Muscle Adaptive Responses to Training) study completed a longitudinal study composed of 12-week high-intensity interval training (HIIT) intervention. Vastus lateralis muscle biopsies were collected at baseline and after 4, 8, and 12 weeks of HIIT. DNA methylation (~850 CpG sites) and proteomic (~3000 proteins) analyses were conducted at all time points. Mixed models were applied to estimate group and individual changes, and methylome and proteome integration was conducted using a holistic multilevel approach with the mixOmics package. A total of 461 proteins significantly changed over time (at 4, 8, and 12 weeks), whilst methylome overall shifted with training only one differentially methylated position (DMP) was significant (adj.p-value < .05). K-means analysis revealed cumulative protein changes by clusters of proteins that presented similar changes over time. Individual responses to training were observed in 101 proteins. Seven proteins had large effect-sizes >0.5, among them are two novel exercise-related proteins, LYRM7 and EPN1. Integration analysis showed bidirectional relationships between the methylome and proteome. We showed a significant influence of HIIT on the epigenome and more so on the proteome in human muscle, and uncovered groups of proteins clustering according to similar patterns across the exercise intervention. Individual responses to exercise were observed in the proteome with novel mitochondrial and metabolic proteins consistently changed across individuals. Future work is required to elucidate the role of these proteins in response to exercise.PMID:37698381 | DOI:10.1096/fj.202300840RR

J Agric Food Chem. 2023 Sep 12. doi: 10.1021/acs.jafc.3c04007. Online ahead of print.ABSTRACTPlants activate direct and indirect defense mechanisms in response to perceived herbivore invasion, which results in negative consequences for herbivores. Tetranychus cinnabarinus is a polyphagous generalist herbivore that inflicts substantial agricultural and horticultural damage. Our study revealed that mite feeding significantly increased jasmonic acid (JA) in the eggplant. The damage inflicted by the mites decreased considerably following the artificial application of JA, thereby indicating that JA initiated the defense response of the eggplant against mites. The transcriptomic and metabolomic analyses demonstrated the activation of the JA-coumarin pathway in response to mite feeding. This pathway protects the eggplant by suppressing the reproductive capacity and population size of the mites. The JA and coumarin treatments suppressed the vitellogenin gene (TcVg6) expression level. Additionally, RNA interference with TcVg6 significantly reduced the egg production and hatching rate of mites. In conclusion, the JA-coumarin pathway in the eggplant decreases the egg-hatching rate of mites through suppression of TcVg6.PMID:37698370 | DOI:10.1021/acs.jafc.3c04007

Biol Rev Camb Philos Soc. 2023 Sep 12. doi: 10.1111/brv.13015. Online ahead of print.ABSTRACTAquatic invertebrates play a pivotal role in (eco)toxicological assessments because they offer ethical, cost-effective and repeatable testing options. Additionally, their significance in the food chain and their ability to represent diverse aquatic ecosystems make them valuable subjects for (eco)toxicological studies. To ensure consistency and comparability across studies, international (eco)toxicology guidelines have been used to establish standardised methods and protocols for data collection, analysis and interpretation. However, the current standardised protocols primarily focus on a limited number of aquatic invertebrate species, mainly from Arthropoda, Mollusca and Annelida. These protocols are suitable for basic toxicity screening, effectively assessing the immediate and severe effects of toxic substances on organisms. For more comprehensive and ecologically relevant assessments, particularly those addressing long-term effects and ecosystem-wide impacts, we recommended the use of a broader diversity of species, since the present choice of taxa exacerbates the limited scope of basic ecotoxicological studies. This review provides a comprehensive overview of (eco)toxicological studies, focusing on major aquatic invertebrate taxa and how they are used to assess the impact of chemicals in diverse aquatic environments. The present work supports the use of a broad-taxa approach in basic environmental assessments, as it better represents the natural populations inhabiting various ecosystems. Advances in omics and other biochemical and computational techniques make the broad-taxa approach more feasible, enabling mechanistic studies on non-model organisms. By combining these approaches with in vitro techniques together with the broad-taxa approach, researchers can gain insights into less-explored impacts of pollution, such as changes in population diversity, the development of tolerance and transgenerational inheritance of pollution responses, the impact on organism phenotypic plasticity, biological invasion outcomes, social behaviour changes, metabolome changes, regeneration phenomena, disease susceptibility and tissue pathologies. This review also emphasises the need for harmonised data-reporting standards and minimum annotation checklists to ensure that research results are findable, accessible, interoperable and reusable (FAIR), maximising the use and reusability of data. The ultimate goal is to encourage integrated and holistic problem-focused collaboration between diverse scientific disciplines, international standardisation organisations and decision-making bodies, with a focus on transdisciplinary knowledge co-production for the One-Health approach.PMID:37698089 | DOI:10.1111/brv.13015

Am J Physiol Lung Cell Mol Physiol. 2023 Sep 12. doi: 10.1152/ajplung.00139.2023. Online ahead of print.ABSTRACTBackground: Antenatal steroid therapy is standard of care for women at imminent risk of preterm delivery. Current dosing regimens employ supra-pharmacological doses to achieve extended fetal steroid exposures. Objective: We aimed to determine the lowest fetal plasma betamethasone concentration sufficient to achieve functional preterm lung maturation. Study design: Ewes with singles fetuses underwent surgery to install a fetal jugular catheter. Adopting a step-wise design, ewes were randomised to either a saline-only group (Negative Control Group; n=9), or one of four betamethasone-treatment groups. Each betamethasone group fetus received a fetal intravenous infusion to target a constant plasma betamethasone levels of either: i) 2 ng/ml (2 ng/ml Positive Control Group, n=9); ii) 1 ng/ml, (1 ng/ml Group, n=10); iii) 0.5 ng/ml (0.5 ng/ml Group, n=10); or iv) 0.25 ng/ml Group (0.25 ng/ml Group, n=10). Fetuses were infused for 48 hours, delivered and ventilated. The Positive Control Group, Negative Control Group and mid-point 0.5 ng/ml Group animals were tested first. An interim analysis informed the final betamethasone group tested. Results: Positive Control Group animals had large, statistically significant improvements in respiratory function. Based on an interim analysis, the 1.0 ng/ml Group was studied in favour of the 0.25 ng/ml Group. Treatment efficacy was progressively lost at plasma betamethasone concentrations lower than 2 ng/ml. Conclusion: We demonstrated that the acute respiratory benefit conveyed by antenatal steroid exposure in the fetal sheep is progressively lost when constant fetal plasma betamethasone concentrations are reduced below a targeted value of 2 ng/ml.PMID:37697929 | DOI:10.1152/ajplung.00139.2023

Analyst. 2023 Sep 12. doi: 10.1039/d3an01021j. Online ahead of print.ABSTRACTSurface-enhanced Raman scattering (SERS) is a powerful technique for detecting trace amounts of analytes. However, the performance of SERS substrates depends on many variables including the enhancement factor, morphology, consistency, and interaction with target analytes. In this study, we investigated, for the first time, the use of electrospray deposition (ESD) combined with a novel ambient focusing DC ion funnel to deposit a high density of gold nanoparticles (AuNPs) to generate large-area, uniform substrates for highly sensitive SERS analysis. We found that the combination of ambient ion focusing with ESD facilitated high-density and intact deposition of non-spherical NPs. This also allowed us to take advantage of a polydisperse colloidal solution of AuNPs (consisting of nanospheres and nanorods), as confirmed by finite-difference time domain (FDTD) simulations. Our SERS substrate exhibited excellent capture capacity for model analyte molecules, namely 4-aminothiophenol (4-ATP) and Rhodamine 6G (R6G), with detection limits in the region of 10-11 M and a relative standard deviation of <6% over a large area (∼500 × 500 μm2). Additionally, we assessed the quantitative performance of our SERS substrate using the R6G probe molecule. The results demonstrated excellent linearity (R2 > 0.99) over a wide concentration range (10-4 M to 10-10 M) with a detection limit of 80 pM.PMID:37697928 | DOI:10.1039/d3an01021j

J Appl Toxicol. 2023 Sep 12. doi: 10.1002/jat.4532. Online ahead of print.ABSTRACTHuobahua, namely, Tripterygium hypoglaucum (Levl.) Hutch, known as a traditional Chinese herbal medicine, especially its underground parts, has been widely developed into several Tripterygium agents for the treatment of rheumatoid arthritis and other autoimmune diseases. It has sparked wide public concern about its safety, such as multi-organ toxicity. However, the toxic characteristics and damage mechanism of Huobahuagen extract (HBHGE) remain unclear. In the present study, subchronic oral toxicity study of HBHGE (10.0 g crude drug/kg/day for 12 weeks) was performed in male rats. Hematological, serum biochemical, and histopathological parameters, urinalysis, and plasma metabolic profiling were assessed. The single-dose subchronic toxicity results related to HBHGE exhibited obvious toxicity to the testis and epididymis of male rats. Furthermore, plasma metabolomics analysis suggested that a series of metabolic disorders were induced by oral administration of HBHGE, mainly focusing on amino acid (glutamate, phenylalanine, and tryptophan) metabolisms, pyrimidine metabolism, glutathione metabolism, and steroid hormone biosynthesis. Moreover, it appeared that serum testosterone in male rats treated with HBHGE for 12 weeks, decreased significantly, and was susceptible to the toxic effects of HBHGE. Taken together, conventional pathology and plasma metabolomics for preliminarily exploring subchronic toxicity and underlying mechanism can provide useful information about the reduction of toxic risks from HBHGE and new insights into the development of detoxification preparations.PMID:37697829 | DOI:10.1002/jat.4532

Hum Reprod. 2023 Sep 11:dead181. doi: 10.1093/humrep/dead181. Online ahead of print.ABSTRACTSTUDY QUESTION: Is the abundance of certain biochemical compounds in human cumulus cells (CCs) related to oocyte quality?SUMMARY ANSWER: Malonate, 5-oxyproline, and erythronate were positively associated with pregnancy potential.WHAT IS KNOWN ALREADY: CCs are removed and discarded prior to ICSI, thereby constituting an interesting biological material on which to perform molecular analysis aimed to predict oocyte developmental competence. Mitochondrial DNA content and transcriptional analyses in CC have been shown to provide a poor predictive value of oocyte competence, but the untargeted analysis of biochemical compounds (metabolomics) has been unexplored.STUDY DESIGN, SIZE, DURATION: CCs were obtained from three groups of cumulus-oocyte complexes (COCs) of known developmental potential: oocytes not developing to blastocyst following ICSI (Bl-); oocytes developing to blastocyst but failing to establish pregnancy following embryo transfer (P-); and oocytes developing to blastocyst able to establish a pregnancy (P+). Metabolomics analyses were performed on 12 samples per group, each sample comprising the CC recovered from a single COC.PARTICIPANTS/MATERIALS, SETTING, METHODS: Human CC samples were obtained from IVF treatments. Only unfrozen oocytes and embryos not submitted to preimplantation genetic testing were included in the analysis. Metabolomics analysis was performed by ultra-high performance liquid chromatography-tandem mass spectroscopy.MAIN RESULTS AND THE ROLE OF CHANCE: The analysis identified 98 compounds, five of which were differentially abundant (P < 0.05) between groups: asparagine, proline, and malonate were less abundant in P- compared to Bl-, malonate and 5-oxoproline were less abundant in P- group compared to P+, and erythronate was less abundant in Bl- group compared to P+. No significant association between the abundance of the compounds identified and donor age or BMI was noted.LIMITATIONS, REASONS FOR CAUTION: Data dispersion and the lack of coherence between developmental groups preclude the direct use of metabolic markers in clinical practice, where the uterine environment plays a major role in pregnancy outcome. The abundance of other compounds not detected by the analysis may be associated with oocyte competence. As donors were lean (only two with BMI > 30 kg/m2) and young (<34 years old), a possible effect of obesity or advanced age on the CC metabolome could not be determined.WIDER IMPLICATIONS OF THE FINDINGS: The abundance of malonate, 5-oxyproline, and erythronate in CC was significantly higher in COCs ultimately establishing pregnancy, providing clues on the pathways required for oocyte competence. The untargeted analysis uncovered the presence of compounds that were not expected in CC, such as β-citrylglutamate and the neurotransmitter N-acetyl-aspartyl-glutamate, which may play roles in chromatin remodeling and signaling, respectively.STUDY FUNDING/COMPETING INTEREST(S): Research was supported by the Industrial Doctorate Project IND2017/BIO-7748 funded by Madrid Region Government. The authors declare no competing interest.TRIAL REGISTRATION NUMBER: N/A.PMID:37697661 | DOI:10.1093/humrep/dead181

Bioinformatics. 2023 Sep 2;39(9):btad450. doi: 10.1093/bioinformatics/btad450.ABSTRACTMOTIVATION: Several applications in constraint-based modelling can be mathematically formulated as cardinality optimization problems involving the minimization or maximization of the number of nonzeros in a vector. These problems include testing for stoichiometric consistency, testing for flux consistency, testing for thermodynamic flux consistency, computing sparse solutions to flux balance analysis problems and computing the minimum number of constraints to relax to render an infeasible flux balance analysis problem feasible. Such cardinality optimization problems are computationally complex, with no known polynomial time algorithms capable of returning an exact and globally optimal solution.RESULTS: By approximating the zero-norm with nonconvex continuous functions, we reformulate a set of cardinality optimization problems in constraint-based modelling into a difference of convex functions. We implemented and numerically tested novel algorithms that approximately solve the reformulated problems using a sequence of convex programs. We applied these algorithms to various biochemical networks and demonstrate that our algorithms match or outperform existing related approaches. In particular, we illustrate the efficiency and practical utility of our algorithms for cardinality optimization problems that arise when extracting a model ready for thermodynamic flux balance analysis given a human metabolic reconstruction.AVAILABILITY AND IMPLEMENTATION: Open source scripts to reproduce the results are here https://github.com/opencobra/COBRA.papers/2023_cardOpt with general purpose functions integrated within the COnstraint-Based Reconstruction and Analysis toolbox: https://github.com/opencobra/cobratoolbox.PMID:37697651 | DOI:10.1093/bioinformatics/btad450

Proteomics Clin Appl. 2023 Sep 11:e2200107. doi: 10.1002/prca.202200107. Online ahead of print.ABSTRACTBACKGROUND: Chronic subdural hematoma (CSDH) is one of the most common neurosurgical diseases with atypical manifestations. The aim of this study was to utilize urine metabolomics to explore potential biomarkers for the diagnosis and prognosis of CSDH.METHODS: Seventy-seven healthy controls and ninety-two patients with CSDH were enrolled in our study. In total, 261 urine samples divided into the discovery group and validation group were analyzed by LC-MS. The statistical analysis and functional annotation were applied to discover potential biomarker panels and altered metabolic pathways.RESULTS: A total of 53 differential metabolites were identified in this study. And the urinary metabolic profiles showed apparent separation between patients and controls. Further functional annotation showed that the differential metabolites were associated with lipid metabolism, fatty acid metabolism, amino acid metabolism, biotin metabolism, steroid hormone biosynthesis, and pentose and glucuronate interconversions. Moreover, one panel of Capryloylglycine, cis-5-Octenoic acid, Ethisterone, and 5,6-DiHETE showed good predictive performance in the diagnosis of CSDH, with an AUC of 0.89 in discovery group and an AUC of 0.822 in validation group. Another five metabolites (Trilobinol, 3'-Hydroxyropivacaine, Ethisterone, Arginyl-Proline, 5-alpha-Dihydrotestosterone glucuronide) showed the levels of them returned to a healthy state after surgery, showing good possibility to monitor the recovery of CSDH patients.CONCLUSION AND CLINICAL RELEVANCE: The findings of the study revealed urine metabolomic differences between CSDH and controls. The potentially diagnostic and prognostic biomarker panels of urine metabolites were established, and functional analysis demonstrated deeper metabolic disorders of CSDH, which might conduce to improve early diagnose of CSDH clinically.PMID:37697649 | DOI:10.1002/prca.202200107

FASEB J. 2023 Oct;37(10):e23185. doi: 10.1096/fj.202300941R.ABSTRACTSensory neurons in the dorsal root ganglia (DRG) convey somatosensory and metabolic cues to the central nervous system and release substances from stimulated terminal endings in peripheral organs. Sex-biased variations driven by the sex chromosome complement (XX and XY) have been implicated in the sensory-islet crosstalk. However, the molecular underpinnings of these male-female differences are not known. Here, we aim to characterize the molecular repertoire and the secretome profile of the lower thoracic spinal sensory neurons and to identify molecules with sex-biased insulin sensing- and/or insulin secretion-modulating activity that are encoded independently of circulating gonadal sex hormones. We used transcriptomics and proteomics to uncover differentially expressed genes and secreted molecules in lower thoracic T5-12 DRG sensory neurons derived from sexually immature 3-week-old male and female C57BL/6J mice. Comparative transcriptome and proteome analyses revealed differential gene expression and protein secretion in DRG neurons in males and females. The transcriptome analysis identified, among others, higher insulin signaling/sensing capabilities in female DRG neurons; secretome screening uncovered several sex-specific candidate molecules with potential regulatory functions in pancreatic β cells. Together, these data suggest a putative role of sensory interoception of insulin in the DRG-islet crosstalk with implications in sensory feedback loops in the regulation of β-cell activity in a sex-biased manner. Finally, we provide a valuable resource of molecular and secretory targets that can be leveraged for understanding insulin interoception and insulin secretion and inform the development of novel studies/approaches to fathom the role of the sensory-islet axis in the regulation of energy balance in males and females.PMID:37695721 | DOI:10.1096/fj.202300941R

Thorax. 2023 Sep 11:thorax-2022-219725. doi: 10.1136/thorax-2022-219725. Online ahead of print.ABSTRACTBACKGROUND: We recently conducted a double-blinded randomised controlled trial showing that fish-oil supplementation during pregnancy reduced the risk of persistent wheeze or asthma in the child by 30%. Here, we explore the mechanisms of the intervention.METHODS: 736 pregnant women were given either placebo or n-3 long-chain polyunsaturated fatty acids (LCPUFAs) in the third trimester in a randomised controlled trial. Deep clinical follow-up of the 695 children in the trial was done at 12 visits until age 6 years, including assessment of genotype at the fatty acid desaturase (FADS) locus, plasma fatty acids, airway DNA methylation, gene expression, microbiome and metabolomics.RESULTS: Supplementation with n-3 LCPUFA reduced the overall risk of non-atopic asthma by 73% at age 6 (relative risk (RR) 0.27 (95% CI 0.06 to 0.85), p=0.042). In contrast, there was no overall effect on asthma with atopic traits (RR 1.42 (95% CI 0.63 to 3.38), p=0.40), but this was significantly modified by maternal FADS genotype and LCPUFA blood levels (interaction p<0.05), and supplementation did reduce the risk of atopic asthma in the subgroup of mothers with FADS risk variants and/or low blood levels of n-3 LCPUFA before the intervention (RR 0.31 (95% CI 0.11 to 0.75), p=0.016). Furthermore, n-3 LCPUFA significantly reduced the number of infections (croup, gastroenteritis, tonsillitis, otitis media and pneumonia) by 16% (incidence rate ratio 0.84 (95% CI 0.74 to 0.96), p=0.009).CONCLUSIONS: n-3 LCPUFA supplementation in pregnancy showed protective effects on non-atopic asthma and infections. Protective effects on atopic asthma depended on maternal FADS genotype and n-3 LCPUFA levels. This indicates that the fatty acid pathway is involved in multiple mechanisms affecting the risk of asthma subtypes and infections.TRIAL REGISTRATION NUMBER: NCT00798226.PMID:37696621 | DOI:10.1136/thorax-2022-219725

Chemosphere. 2023 Sep 9:140120. doi: 10.1016/j.chemosphere.2023.140120. Online ahead of print.ABSTRACTEmerging epidemiological evidence indicates potential associations between gestational perfluorobutane sulfonate (PFBS) exposure and adverse metabolic outcomes in offspring. However, the underlying mechanisms remain unclear. Our study aimed to investigate PFBS exposure effects during pregnancy and lactation on rat offspring lipid profiles and the possible underlying mechanisms. Although the biochemical index difference including total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), alanine amino transaminase (ALT), aspartate amino transferase (AST), and fasting blood glucose between exposed groups and the control group was not significant, transcriptome analyses showed that the differentially expressed genes (DEGs) in the 50 mg/kg/day PFBS exposure group were significantly related to protein digestion and absorption, peroxisome proliferator activated-receptor (PPAR) signaling pathway, xenobiotic metabolism by cytochrome P450, glycine, serine and threonine metabolism, β-alanine metabolism, bile secretion, unsaturated fatty acid (FA) biosynthesis, and alanine, aspartate and glutamate metabolism. Untargeted metabolomics analyses identified 17 differential metabolites in the 50 mg/kg/day PFBS exposure group. Among these, phosphatidylserine [PS (18:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z))], lysoPE (18:1(11Z)/0:0), and PS (14:0/20:4(5Z,8Z,11Z,14Z)) were significantly correlated with phospholipid metabolism disorders. Correlation analysis indicated the DEGs, including FA binding protein (Fabp4), spermine oxidase (Smox), Fabp2, acyl-CoA thioesterase 5 (Acot5), sarcosine dehydrogenase (Sardh), and amine oxidase, copper-containing 3 (Aoc3) that significantly enriched in xenobiotic metabolism by cytochrome P450 and glycine, serine, and threonine metabolism signaling pathways were highly related to the differential metabolite pantetheine 4'-phosphate. Pantetheine 4'-phosphate was significantly negatively associated with non-high-density lipoprotein (non-HDL) and TC levels. Collectively, our study indicated that maternal PFBS exposure at a relatively low level could alter gene expression and metabolic molecules in lipid metabolism-related pathway series in rat offspring, although the effects on metabolic phenotypes were not significant within the limited observational period, using group-wise and trend analyses.PMID:37696479 | DOI:10.1016/j.chemosphere.2023.140120