PubMed

Plants (Basel). 2023 Oct 1;12(19):3461. doi: 10.3390/plants12193461.ABSTRACT(1) Background: Cytoplasmic male sterility (CMS) is important for exploiting heterosis. Soybean (Glycine max L.) has a low outcrossing rate that is detrimental for breeding sterile lines and producing hybrid seeds. Therefore, the molecular mechanism controlling the outcrossing rate should be elucidated to increase the outcrossing rate of soybean CMS lines; (2) Methods: The male-sterile soybean lines JLCMS313A (with a high outcrossing rate; HL) and JLCMS226A (with a low outcrossing rate; LL) were used for a combined analysis of the transcriptome (RNA-seq) and the targeted phenol metabolome; (3) Results: The comparison between HL and LL detected 5946 differentially expressed genes (DEGs) and 81 phenolic metabolites. The analysis of the DEGs and differentially abundant phenolic metabolites identified only one common KEGG pathway related to flavonoid biosynthesis. The qRT-PCR expression for eight DEGs was almost consistent with the transcriptome data. The comparison of the cloned coding sequence (CDS) regions of the SUS, FLS, UGT, and F3H genes between HL and LL revealed seven single nucleotide polymorphisms (SNPs) only in the F3H CDS. Moreover, five significant differentially abundant phenolic metabolites between HL and LL were associated with flavonoid metabolic pathways. Finally, on the basis of the SNPs in the F3H CDS, one derived cleaved amplified polymorphic sequence (dCAPS) marker was developed to distinguish between HL and LL soybean lines; (4) Conclusions: The flavonoid biosynthesis pathway may indirectly affect the outcrossing rate of CMS sterile lines in soybean.PMID:37836201 | DOI:10.3390/plants12193461

Plants (Basel). 2023 Sep 22;12(19):3359. doi: 10.3390/plants12193359.ABSTRACTRice is a crucial global food crop, but it lacks a natural tolerance to high salt levels, resulting in significant yield reductions. To gain a comprehensive understanding of the molecular mechanisms underlying rice's salt tolerance, further research is required. In this study, the transcriptomic and metabolomic differences between the salt-tolerant rice variety Lianjian5 (TLJIAN) and the salt-sensitive rice variety Huajing5 (HJING) were examined. Transcriptome analysis revealed 1518 differentially expressed genes (DEGs), including 46 previously reported salt-tolerance-related genes. Notably, most of the differentially expressed transcription factors, such as NAC, WRKY, MYB, and EREBP, were upregulated in the salt-tolerant rice. Metabolome analysis identified 42 differentially accumulated metabolites (DAMs) that were upregulated in TLJIAN, including flavonoids, pyrocatechol, lignans, lipids, and trehalose-6-phosphate, whereas the majority of organic acids were downregulated in TLJIAN. The interaction network of 29 differentially expressed transporter genes and 19 upregulated metabolites showed a positive correlation between the upregulated calcium/cation exchange protein genes (OsCCX2 and CCX5_Ath) and ABC transporter gene AB2E_Ath with multiple upregulated DAMs in the salt-tolerant rice variety. Similarly, in the interaction network of differentially expressed transcription factors and 19 upregulated metabolites in TLJIAN, 6 NACs, 13 AP2/ERFs, and the upregulated WRKY transcription factors were positively correlated with 3 flavonoids, 3 lignans, and the lipid oleamide. These results suggested that the combined effects of differentially expressed transcription factors, transporter genes, and DAMs contribute to the enhancement of salt tolerance in TLJIAN. Moreover, this study provides a valuable gene-metabolite network reference for understanding the salt tolerance mechanism in rice.PMID:37836098 | DOI:10.3390/plants12193359

Animals (Basel). 2023 Sep 30;13(19):3076. doi: 10.3390/ani13193076.ABSTRACTThe hair follicle (HF) is the fundamental unit for fleece and cashmere production in cashmere goats and is crucial in determining cashmere yield and quality. The mechanisms regulating HF development in cashmere goats during the embryonic period remain unclear. Growing evidence suggests that HF development involves complex developmental stages and critical events, and identifying the underlying factors can improve our understanding of HF development. In this study, samples were collected from embryonic day 75 (E75) to E125, the major HF developmental stages. The embryonic HFs of cashmere goats were subjected to proteomic and metabolomic analyses, which revealed dynamic changes in the key factors and signalling pathways controlling HF development at the protein and metabolic levels. Gene ontology and the Kyoto Encyclopaedia of Genes and Genomes were used to functionally annotate 1784 significantly differentially expressed proteins and 454 significantly differentially expressed metabolites enriched in different HF developmental stages. A joint analysis revealed that the oxytocin signalling pathway plays a sustained role in embryonic HF development by activating the MAPK and Ca2+ signalling pathways, and a related regulatory network map was constructed. This study provides a global perspective on the mechanism of HF development in cashmere goats and enriches our understanding of embryonic HF development.PMID:37835682 | DOI:10.3390/ani13193076

Animals (Basel). 2023 Sep 27;13(19):3034. doi: 10.3390/ani13193034.ABSTRACTThe gut microbiota plays an important role in the physiological activities of the host and affects the formation of important economic traits in livestock farming. The effects of cecal microbiota on chicken weights were investigated using the Guizhou yellow chicken as a model. Experimental cohorts from chickens with high- (HC, n = 16) and low-market-weights (LC, n = 16) were collected. Microbial 16S rRNA gene sequencing and non-targeted serum metabolome data were integrated to explore the effect and metabolic mechanism of cecal microbiota on market weight. The genera Lachnoclostridium, Alistipes, Negativibacillus, Sellimonas, and Ruminococcus torques were enriched in the HC group, while Phascolarctobacterium was enriched in the LC group (p < 0.05). Metabolomic analysis determined that pantothenic acid (vitamin B5), luvangetin (2H-1-benzopyran-6-acrylic acid), and menadione (vitamin K3) were significantly higher in HC serum, while beclomethasone dipropionate (a glucocorticoid) and chlorophene (2-benzyl-4-chlorophenol) were present at higher levels in the LC group. The microbes enriched in HC were significantly positively correlated with metabolites, including pantothenic acid and menadione, and negatively correlated with beclomethasone dipropionate and chlorophene. These results indicated that specific cecal bacteria in Guizhou yellow chickens alter the host metabolism and growth performance. This study provides a reference for revealing the mechanism of cecal microbe actions that affect chicken body weight.PMID:37835639 | DOI:10.3390/ani13193034

Foods. 2023 Oct 9;12(19):3708. doi: 10.3390/foods12193708.ABSTRACTScenting tea with Jasminum sambac is beneficial to forming a unique taste of jasmine tea, which is regulated by numerous compounds. To investigate the relationship between metabolites in jasmine and jasmine tea, as well as the impact of metabolites on the characteristic taste of jasmine tea, the liquid chromatography-mass spectrometry, sensory evaluation, and multivariate analysis were applied in this study. A total of 585 and 589 compounds were identified in jasmine tea and jasmine, respectively. After scented, jasmine tea added 70 compounds, which were believed to come from jasmine flowers. Furthermore, seventy-four compounds were identified as key characteristic compounds of jasmine tea, and twenty-two key differential metabolite compounds were believed to be used to distinguish jasmine tea scented differently and contribute to the taste of jasmine tea. Additionally, the relationship between taste compounds and aroma quality was also explored, and it was found that five compounds were positively correlated with the aroma properties of jasmine tea and seven compounds were negatively correlated with the aroma properties of jasmine tea. Overall, these findings provided insights into the future study of the mechanism of taste formation in jasmine tea and provided the theoretical basis for the production of jasmine tea.PMID:37835360 | DOI:10.3390/foods12193708

Foods. 2023 Oct 9;12(19):3704. doi: 10.3390/foods12193704.ABSTRACTTraditional fermented milk from the western Sichuan plateau of China has a unique flavor and rich microbial diversity. This study explored the quality formation mechanism in fermented milk inoculated with Lactobacillus brevis NZ4 and Kluyveromyces marxianus SY11 (MFM), the dominant microorganisms isolated from traditional dairy products in western nan. The results indicated that MFM displayed better overall quality than the milk fermented with L. brevis NZ4 (LFM) and K. marxianus SY11 (KFM), respectively. MFM exhibited good sensory quality, more organic acid types, more free amino acids and esters, and moderate acidity and ethanol concentrations. Non-targeted metabolomics showed a total of 885 metabolites annotated in the samples, representing 204 differential metabolites between MFM and LFM and 163 between MFM and KFM. MFM displayed higher levels of N-acetyl-L-glutamic acid, cysteinyl serine, glaucarubin, and other substances. The differential metabolites were mainly enriched in pathways such as glycerophospholipid metabolism, arginine biosynthesis, and beta-alanine metabolism. This study speculated that L. brevis affected K. marxianus growth via its metabolites, while the mixed fermentation of these strains significantly changed the metabolism pathway of flavor-related substances, especially glycerophospholipid metabolism. Furthermore, mixed fermentation modified the flavor and quality of fermented milk by affecting cell growth and metabolic pathways.PMID:37835356 | DOI:10.3390/foods12193704

Foods. 2023 Sep 28;12(19):3608. doi: 10.3390/foods12193608.ABSTRACTSWATHtoMRM technology was used in this experiment to further identify and trace the sources of Dendrobium devonianum and Dendrobium officinale produced in the same area using TOF and MS-MRM. After the conversion of the R package of SWATHtoMRM, 191 MRM pairs of positive ions and 96 pairs of negative ions were obtained. Dendrobium devonianum and Dendrobium officinale can be separated very well using the PCA and PLS-DA analysis of MRM ion pairs; this shows that there are obvious differences in chemical composition between Dendrobium devonianum and Dendrobium officinale, which clearly proves that the pseudotargeted metabolomics method based on SWATHtoMRM can be used for traceability identification research. A total of 146 characteristic compounds were obtained, with 20 characteristic compounds in Dendrobium devonianum. The enrichment pathways of the characteristic compounds were mainly concentrated in lipids and atherosclerosis, chagas disease, fluid shear stress and atherosclerosis, proteoglycans in cancer, the IL-17 signaling pathway, the sphingolipid signaling pathway, diabetic cardiomyopathy, arginine and proline metabolism, etc., among which the lipid and atherosclerosis pathways were more enriched, and 11 characteristic compounds affected the expression levels of IL-1, TNFα, CD36, IL-1β, etc. These can be used as a reference for research on variety improvement and active substance accumulation in Dendrobium devonianum and Dendrobium officinale.PMID:37835262 | DOI:10.3390/foods12193608

Foods. 2023 Sep 26;12(19):3579. doi: 10.3390/foods12193579.ABSTRACTInactivation is a crucial step in the production of postbiotics, with thermal inactivation being the prevailing method employed. Nevertheless, the impact of thermal treatment on bioactivity and chemical composition remains unexplored. The objective of this study was to assess the influence of heating temperature on the antioxidant, anti-inflammatory properties and the chemical composition of ET-22 and BL-99 postbiotics. The findings revealed that subjecting ET-22 and BL-99 to thermal treatment ranging from 70 °C to 121 °C for a duration of 10 min effectively deactivated them, leading to the disruption of cellular structure and release of intracellular contents. The antioxidant and anti-inflammatory activity of ET-22 and BL-99 postbiotics remained unaffected by mild heating temperatures (below 100 °C). However, excessive heating at 121 °C diminished the antioxidant activity of the postbiotic. To further investigate the impact of thermal treatments on chemical composition, non-targeted metabolomics was conducted to analyze the cell-free supernatants derived from ET-22 and BL-99. The results revealed that compared to mild inactivation at temperatures below 100 °C, the excessive temperature of 121 °C significantly altered the chemical profile of the postbiotic. Several bioactive components with antioxidant and anti-inflammatory properties, including zomepirac, flumethasone, 6-hydroxyhexanoic acid, and phenyllactic acid, exhibited a significant reduction in their levels following exposure to a temperature of 121 °C. This decline in their abundance may be associated with a corresponding decrease in their antioxidant and anti-inflammatory activities. The cumulative evidence gathered strongly indicates that heating temperatures exert a discernible influence on the properties of postbiotics, whereby excessive heating leads to the degradation of heat-sensitive active constituents and subsequent diminishment of their biological efficacy.PMID:37835233 | DOI:10.3390/foods12193579

Foods. 2023 Sep 24;12(19):3547. doi: 10.3390/foods12193547.ABSTRACTGrass-finished beef (GFB) can provide beneficial bioactive compounds to healthy diets, including omega-3 polyunsaturated fatty acids (n-3 PUFAs), conjugated linoleic acid (CLA), and secondary bioactive compounds, such as phytochemicals. The objective of this study was to compare fatty acids (FAs), micronutrients, and phytochemicals of beef fed a biodiverse pasture (GRASS), a total mixed ration (GRAIN), or a total mixed ration with 5% grapeseed extract (GRAPE). This was a two-year study involving fifty-four Red Angus steers (n = 54). GFB contained higher levels of n-3 PUFAs, vitamin E, iron, zinc, stachydrine, hippuric acid, citric acid, and succinic acid than beef from GRAIN and GRAPE (p < 0.001 for all). No differences were observed in quantified phytochemicals between beef from GRAIN and GRAPE (p > 0.05). Random forest analysis indicated that phytochemical and FA composition of meat can predict cattle diets with a degree of certainty, especially for GFB (5.6% class error). In conclusion, these results indicate that GFB contains higher levels of potentially beneficial bioactive compounds, such as n-3 PUFAs, micronutrients, and phytochemicals, compared to grain-finished beef. Additionally, the n-6:n-3 ratio was the most crucial factor capable of separating beef based on finishing diets.PMID:37835200 | DOI:10.3390/foods12193547

Foods. 2023 Sep 22;12(19):3537. doi: 10.3390/foods12193537.ABSTRACTIn order to analyze the changes in the microbial community structure during the pile fermentation of Qingzhuan tea and their correlation with the formation of quality compounds in Qingzhuan tea, this study carried out metagenomic and metabolomic analyses of tea samples during the fermentation process of Qingzhuan tea. The changes in the expression and abundance of microorganisms during the pile fermentation were investigated through metagenomic assays. During the processing of Qingzhuan tea, there is a transition from a bacterial dominated ecosystem to an ecosystem enriched with fungi. The correlation analyses of metagenomics and metabolomics showed that amino acids and polyphenol metabolites with relatively simple structures exhibited a significant negative correlation with target microorganisms, while the structurally complicated B-ring dihydroxy puerin, B-ring trihydroxy galloyl puerlin, and other compounds showed a significant positive correlation with target microorganisms. Aspergillus niger, Aspergillus glaucus, Penicillium in the Aspergillaceae family, and Talaromyces and Rasamsonia emersonii in Trichocomaceae were the key microorganisms involved in the formation of the characteristic qualities of Qingzhuan tea.PMID:37835190 | DOI:10.3390/foods12193537

Foods. 2023 Sep 22;12(19):3520. doi: 10.3390/foods12193520.ABSTRACTMicrobial contamination affects the quality of the fermented Houttuynia cordata Thunb. (H. cordata) beverage (FHB). The present study aimed to assess the bio-preservative property of Enterococcus faecium OV3-6 (E. faecium OV3-6) during the production of FHB. The antimicrobial activity against Escherichia coli, Salmonella, Bacillus cereus, and Staphylococcus aureus and the survival of E. faecium OV3-6 were studied. Then, FHB fermentation was performed with different preservatives (non-preservative, E. faecium OV3-6, cell-free supernatant of E. faecium OV3-6, and nisin) with and without representative pathogens. The maximum antimicrobial activity against S. aureus and B. cereus was observed after 18 h of cultivation in an MRS medium. E. faecium OV3-6 was used as a starter to produce the FHB, and the strain survived up to 48 h in the fermented beverage. E. faecium OV3-6 and its cell-free supernatant inhibited the growth of E. coli, Salmonella, B. cereus, and S. aureus in the stimulated FHB. The non-preservatives and nisin-containing FHB showed inhibition against Gram-positive pathogens. The FHB treated with E. faecium OV3-6 was rich in lactic acid bacteria, and the product was at an acceptable level of pH (less than 4.3). Certain limitations were identified in the study, such as lack of nutritional, metabolomics analysis, and safety and consumer acceptability of FHB. The results suggested that E. faecium OV3-6 could be used as a bio-preservative to produce fermented plant beverages (FPBs).PMID:37835173 | DOI:10.3390/foods12193520

J Clin Med. 2023 Sep 27;12(19):6225. doi: 10.3390/jcm12196225.ABSTRACTBACKGROUND: Severe coronavirus disease 2019 (COVID-19) disease courses are characterized by immuno-inflammatory, thrombotic, and parenchymal alterations. Prediction of individual COVID-19 disease courses to guide targeted prevention remains challenging. We hypothesized that a distinct serologic signature precedes surges of IL-6/D-dimers in severely affected COVID-19 patients.METHODS: We performed longitudinal plasma profiling, including proteome, metabolome, and routine biochemistry, on seven seropositive, well-phenotyped patients with severe COVID-19 referred to the Intensive Care Unit at the German Heart Center. Patient characteristics were: 65 ± 8 years, 29% female, median CRP 285 ± 127 mg/dL, IL-6 367 ± 231 ng/L, D-dimers 7 ± 10 mg/L, and NT-proBNP 2616 ± 3465 ng/L.RESULTS: Based on time-series analyses of patient sera, a prediction model employing feature selection and dimensionality reduction through least absolute shrinkage and selection operator (LASSO) revealed a number of candidate proteins preceding hyperinflammatory immune response (denoted ΔIL-6) and COVID-19 coagulopathy (denoted ΔD-dimers) by 24-48 h. These candidates are involved in biological pathways such as oxidative stress/inflammation (e.g., IL-1alpha, IL-13, MMP9, C-C motif chemokine 23), coagulation/thrombosis/immunoadhesion (e.g., P- and E-selectin), tissue repair (e.g., hepatocyte growth factor), and growth factor response/regulatory pathways (e.g., tyrosine-protein kinase receptor UFO and low-density lipoprotein receptor (LDLR)). The latter are host- or co-receptors that promote SARS-CoV-2 entry into cells in the absence of ACE2.CONCLUSIONS: Our novel prediction model identified biological and regulatory candidate networks preceding hyperinflammation and coagulopathy, with the most promising group being the proteins that explain changes in D-dimers. These biomarkers need validation. If causal, our work may help predict disease courses and guide personalized treatment for COVID-19.PMID:37834869 | DOI:10.3390/jcm12196225

Int J Mol Sci. 2023 Oct 9;24(19):15014. doi: 10.3390/ijms241915014.ABSTRACTAutophagy is an evolutionarily conserved mechanism for degrading and recycling various cellular components, functioning in both normal development and stress conditions. This process is tightly regulated by a set of autophagy-related (ATG) proteins, including ATG2 in the ATG9 cycling system and ATG5 in the ATG12 conjugation system. Our recent research demonstrated that autophagy-mediated compartmental cytoplasmic deletion is essential for pollen germination. However, the precise mechanisms through which autophagy regulates pollen germination, ensuring its fertility, remain largely unknown. Here, we applied multi-omics analyses, including transcriptomic and metabolomic approaches, to investigate the downstream pathways of autophagy in the process of pollen germination. Although ATG2 and ATG5 play similar roles in regulating pollen germination, high-throughput transcriptomic analysis reveals that silencing ATG5 has a greater impact on the transcriptome than silencing ATG2. Cross-comparisons of transcriptome and proteome analysis reveal that gene expression at the mRNA level and protein level is differentially affected by autophagy. Furthermore, high-throughput metabolomics analysis demonstrates that pathways related to amino acid metabolism and aminoacyl-tRNA biosynthesis were affected by both ATG2 and ATG5 silencing. Collectively, our multi-omics analyses reveal the central role of autophagy in cellular metabolism, which is critical for initiating pollen germination and ensuring pollen fertility.PMID:37834462 | DOI:10.3390/ijms241915014

Int J Mol Sci. 2023 Oct 9;24(19):15010. doi: 10.3390/ijms241915010.ABSTRACTSaliva houses over 2000 proteins and peptides with poorly clarified functions, including proline-rich proteins, statherin, P-B peptides, histatins, cystatins, and amylases. Their genes are poorly conserved across related species, reflecting an evolutionary adaptation. We searched the nucleotide substitutions fixed in these salivary proteins' gene loci in modern humans compared with ancient hominins. We mapped 3472 sequence variants/nucleotide substitutions in coding, noncoding, and 5'-3' untranslated regions. Despite most of the detected variations being within noncoding regions, the frequency of coding variations was far higher than the general rate found throughout the genome. Among the various missense substitutions, specific substitutions detected in PRB1 and PRB2 genes were responsible for the introduction/abrogation of consensus sequences recognized by convertase enzymes that cleave the protein precursors. Overall, these changes that occurred during the recent human evolution might have generated novel functional features and/or different expression ratios among the various components of the salivary proteome. This may have influenced the homeostasis of the oral cavity environment, possibly conditioning the eating habits of modern humans. However, fixed nucleotide changes in modern humans represented only 7.3% of all the substitutions reported in this study, and no signs of evolutionary pressure or adaptative introgression from archaic hominins were found on the tested genes.PMID:37834461 | DOI:10.3390/ijms241915010

Int J Mol Sci. 2023 Oct 9;24(19):15013. doi: 10.3390/ijms241915013.ABSTRACTSugar beet is susceptible to Beet curly top virus (BCTV), which significantly reduces yield and sugar production in the semi-arid growing regions worldwide. Sources of genetic resistance to BCTV is limited and control depends upon insecticide seed treatments with neonicotinoids. Through double haploid production and genetic selection, BCTV resistant breeding lines have been developed. Using BCTV resistant (R) [KDH13; Line 13 and KDH4-9; Line 4] and susceptible (S) [KDH19-17; Line 19] lines, beet leafhopper mediated natural infection, mRNA/sRNA sequencing, and metabolite analyses, potential mechanisms of resistance against the virus and vector were identified. At early infection stages (2- and 6-days post inoculation), examples of differentially expressed genes highly up-regulated in the 'R' lines (vs. 'S') included EL10Ac5g10437 (inhibitor of trypsin and hageman factor), EL10Ac6g14635 (jasmonate-induced protein), EL10Ac3g06016 (ribosome related), EL10Ac2g02812 (probable prolyl 4-hydroxylase 10), etc. Pathway enrichment analysis showed differentially expressed genes were predominantly involved with peroxisome, amino acids metabolism, fatty acid degradation, amino/nucleotide sugar metabolism, etc. Metabolite analysis revealed significantly higher amounts of specific isoflavonoid O-glycosides, flavonoid 8-C glycosides, triterpenoid, and iridoid-O-glycosides in the leaves of the 'R' lines (vs. 'S'). These data suggest that a combination of transcriptional regulation and production of putative antiviral metabolites might contribute to BCTV resistance. In addition, genome divergence among BCTV strains differentially affects the production of small non-coding RNAs (sncRNAs) and small peptides which may potentially affect pathogenicity and disease symptom development.PMID:37834460 | DOI:10.3390/ijms241915013

Int J Mol Sci. 2023 Oct 8;24(19):14986. doi: 10.3390/ijms241914986.ABSTRACTIn the face of escalating environmental challenges, understanding the intricate relationship between plant metabolites, pollution stress, and climatic conditions is of paramount importance. This study aimed to conduct a comprehensive analysis of metabolic variations generated through 1H and 13C NMR measurements in evergreen needles collected from different regions with varying pollution levels. Multivariate analyses were employed to identify specific metabolites responsive to pollution stress and climatic factors. Air pollution indicators were assessed through ANOVA and Pearson correlation analyses. Our results revealed significant metabolic changes attributed to geographical origin, establishing these conifer species as potential indicators for both air pollution and climatic conditions. High levels of air pollution correlated with increased glucose and decreased levels of formic acid and choline. Principal component analysis (PCA) unveiled a clear species separation, largely influenced by succinic acid and threonine. Discriminant analysis (DA) confirmed these findings, highlighting the positive correlation of glucose with pollution grade. Beyond pollution assessment, these metabolic variations could have ecological implications, impacting interactions and ecological functions. Our study underscores the dynamic interplay between conifer metabolism, environmental stressors, and ecological systems. These findings not only advance environmental monitoring practices but also pave the way for holistic research encompassing ecological and physiological dimensions, shedding light on the multifaceted roles of metabolites in conifer responses to environmental challenges.PMID:37834434 | DOI:10.3390/ijms241914986

Int J Mol Sci. 2023 Oct 6;24(19):14957. doi: 10.3390/ijms241914957.ABSTRACTThioesters of coenzyme A (CoA) carrying different acyl chains (acyl-CoAs) are central intermediates of many metabolic pathways and donor molecules for protein lysine acylation. Acyl-CoA species largely differ in terms of cellular concentrations and physico-chemical properties, rendering their analysis challenging. Here, we compare several approaches to quantify cellular acyl-CoA concentrations in normal and ischemic rat liver, using HPLC and LC-MS/MS for multi-acyl-CoA analysis, as well as NMR, fluorimetric and spectrophotometric techniques for the quantification of acetyl-CoAs. In particular, we describe a simple LC-MS/MS protocol that is suitable for the relative quantification of short and medium-chain acyl-CoA species. We show that ischemia induces specific changes in the short-chain acyl-CoA relative concentrations, while mild ischemia (1-2 min), although reducing succinyl-CoA, has little effects on acetyl-CoA, and even increases some acyl-CoA species upstream of the tricarboxylic acid cycle. In contrast, advanced ischemia (5-6 min) also reduces acetyl-CoA levels. Our approach provides the keys to accessing the acyl-CoA metabolome for a more in-depth analysis of metabolism, protein acylation and epigenetics.PMID:37834405 | DOI:10.3390/ijms241914957

Int J Mol Sci. 2023 Oct 5;24(19):14937. doi: 10.3390/ijms241914937.ABSTRACTBreast cancer is the most prevalent form of cancer among women. The microenvironment of a cancer tumor is surrounded by various cells, including the microbiota. An imbalance between microbes and their host may contribute to the development and spread of breast cancer. Therefore, the objective of this study is to investigate the influence of Enterococcus faecalis on a breast cancer cell line (MCF-7) to mimic the luminal A subtype of breast cancer, using an untargeted proteomics approach to analyze the proteomic profiles of breast cancer cells after their treatment with E. faecalis in order to understand the microbiome and its role in the development of cancer. The breast cancer cell line MCF-7 was cultured and then treated with a 10% bacterial supernatant at two time points (24 h and 48 h) at 37 °C in a humidified incubator with 5% CO2. Proteins were then extracted and separated using two-dimensional difference (2D-DIGE) gel electrophoresis, and the statistically significant proteins (p-value < 0.05, fold change > 1.5) were identified via matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). The protein fingerprints showed a differential protein expression pattern in the cells treated with E. faecalis for 24 and 48 h compared with the control. We found 58 statistically significant proteins changes in the MCF-7 breast cancer cells affected by E. faecalis. Kilin and transgelin were upregulated after 24 h of treatment and could be used as diagnostic and prognostic markers for breast cancer. In addition, another protein involved in the inhibition of cell proliferation was coiled-coil domain-containing protein 154. The protein markers identified in this study may serve as possible biomarkers for breast cancer progression. This promotes their future uses as important therapeutic goals in the treatment and diagnosis of cancer and increases our understanding of the breast microbiome and its role in the development of cancer.PMID:37834385 | DOI:10.3390/ijms241914937

Int J Mol Sci. 2023 Oct 3;24(19):14856. doi: 10.3390/ijms241914856.ABSTRACTPlateau adaptation in animals involves genetic mechanisms as well as coevolutionary mechanisms of the microbiota and metabolome of the animal. Therefore, the characteristics of the rumen microbiome and metabolome, transcriptome, and serum metabolome of Tibetan sheep at different altitudes (4500 m, 3500 m, and 2500 m) were analyzed. The results showed that the rumen differential metabolites at 3500 m and 4500 m were mainly enriched in amino acid metabolism, lipid metabolism, and carbohydrate metabolism, and there was a significant correlation with microbiota. The differentially expressed genes and metabolites at middle and high altitudes were coenriched in asthma, arachidonic acid metabolism, and butanoate and propanoate metabolism. In addition, the serum differential metabolites at 3500 m and 4500 m were mainly enriched in amino acid metabolism, lipid metabolism, and metabolism of xenobiotics by cytochrome P450, and they were also related to microbiota. Further analysis revealed that rumen metabolites accounted for 7.65% of serum metabolites. These common metabolites were mainly enriched in metabolic pathways and were significantly correlated with host genes (p < 0.05). This study found that microbiota, metabolites, and epithelial genes were coenriched in pathways related to lipid metabolism, energy metabolism, and immune metabolism, which may be involved in the regulation of Tibetan sheep adaptation to plateau environmental changes.PMID:37834304 | DOI:10.3390/ijms241914856

Int J Mol Sci. 2023 Oct 2;24(19):14844. doi: 10.3390/ijms241914844.ABSTRACTApolipoprotein-CIII (apo-CIII) is involved in triglyceride-rich lipoprotein metabolism and linked to beta-cell damage, insulin resistance, and cardiovascular disease. Apo-CIII exists in four main proteoforms: non-glycosylated (apo-CIII0a), and glycosylated apo-CIII with zero, one, or two sialic acids (apo-CIII0c, apo-CIII1 and apo-CIII2). Our objective is to determine how apo-CIII glycosylation affects lipid traits and type 2 diabetes prevalence, and to investigate the genetic basis of these relations with a genome-wide association study (GWAS) on apo-CIII glycosylation. We conducted GWAS on the four apo-CIII proteoforms in the DiaGene study in people with and without type 2 diabetes (n = 2318). We investigated the relations of the identified genetic loci and apo-CIII glycosylation with lipids and type 2 diabetes. The associations of the genetic variants with lipids were replicated in the Diabetes Care System (n = 5409). Rs4846913-A, in the GALNT2-gene, was associated with decreased apo-CIII0a. This variant was associated with increased high-density lipoprotein cholesterol and decreased triglycerides, while high apo-CIII0a was associated with raised high-density lipoprotein-cholesterol and triglycerides. Rs67086575-G, located in the IFT172-gene, was associated with decreased apo-CIII2 and with hypertriglyceridemia. In line, apo-CIII2 was associated with low triglycerides. On a genome-wide scale, we confirmed that the GALNT2-gene plays a major role i O-glycosylation of apolipoprotein-CIII, with subsequent associations with lipid parameters. We newly identified the IFT172/NRBP1 region, in the literature previously associated with hypertriglyceridemia, as involved in apolipoprotein-CIII sialylation and hypertriglyceridemia. These results link genomics, glycosylation, and lipid metabolism, and represent a key step towards unravelling the importance of O-glycosylation in health and disease.PMID:37834292 | DOI:10.3390/ijms241914844