Lipid Degradation Research Articles

Influence of Drying and Storage Conditions on the Volatile Organic Compounds Profile of Spirulina Platensis

Spirulina platensis (SP) has gained popularity over the last few years, owing to its remarkable nutritional properties and high potential across various industrial sectors. In this study, we analyzed the volatile profile of eight SP samples from the same strain subjected to different drying (oven-drying, air-drying, and spray-drying) and storing conditions (“freshly prepared” and after 12 months of storage) using HS-SPME-GC-MS. Principal component analysis (PCA) was used as a multivariate technique to discern similarities and differences among the samples. The main aim was to assess the impact of the drying technique on the aroma profile and storage life of SP samples. Air-drying leads to the less pronounced formation of by-products related to heat treatment, such as Maillard and Strecker degradation compounds, but promotes oxidative and fermentative phenomena, with the formation of organic acids and esters, especially during storage. Thermal treatment, essential for limiting degradation and fermentation during storage and extending shelf life, alters the aroma profile through the formation of volatile compounds, such as Strecker aldehydes and linear aldehydes, from amino acid and lipid degradation. High temperatures in spray-drying favor the formation of pyrazines. The findings underscore the trade-offs inherent in choosing an appropriate drying method, thereby informing decision-making processes in industrial settings aimed at optimizing both product quality and efficiency.

Dynamic Interplay between Microbiota Shifts and Differential Metabolites during Dairy Processing and Storage.

Due to the intricate complexity of the original microbiota, residual heat-resistant enzymes, and chemical components, identifying the essential factors that affect dairy quality using traditional methods is challenging. In this study, raw milk, pasteurized milk, and ultra-heat-treated (UHT) milk samples were collectively analyzed using metagenomic next-generation sequencing (mNGS), high-throughput liquid chromatography-mass spectrometry (LC-MS), and gas chromatography-mass spectrometry (GC-MS). The results revealed that raw milk and its corresponding heated dairy products exhibited different trends in terms of microbiota shifts and metabolite changes during storage. Via the analysis of differences in microbiota and correlation analysis of the microorganisms present in differential metabolites in refrigerated pasteurized milk, the top three differential microorganisms with increased abundance, Microbacterium (p < 0.01), unclassified Actinomycetia class (p < 0.05), and Micrococcus (p < 0.01), were detected; these were highly correlated with certain metabolites in pasteurized milk (r > 0.8). This indicated that these genera were the main proliferating microorganisms and were the primary genera involved in the metabolism of pasteurized milk during refrigeration-based storage. Microorganisms with decreased abundance were classified into two categories based on correlation analysis with certain metabolites. It was speculated that the heat-resistant enzyme system of a group of microorganisms with high correlation (r > 0.8), such as Pseudomonas and Acinetobacter, was the main factor causing milk spoilage and that the group with lower correlation (r < 0.3) had a lower impact on the storage process of pasteurized dairy products. By comparing the metabolic pathway results based on metagenomic and metabolite annotation, it was proposed that protein degradation may be associated with microbial growth, whereas lipid degradation may be linked to raw milk's initial heat-resistant enzymes. By leveraging the synergy of metagenomics and metabolomics, the interacting factors determining the quality evolution of dairy products were systematically investigated, providing a novel perspective for controlling dairy processing and storage effectively.

Detection of aldehydes from degradation of lipid nanoparticle formulations using a hierarchically-organized nanopore electrochemical biosensor

Degradation of ionizable lipids in mRNA-based vaccines was recently found to deactivate the payload, demanding rigorous monitoring of impurities in lipid nanoparticle (LNP) formulations. However, parallel screening for lipid degradation in customized delivery systems for next-generation therapeutics maintains a challenging and unsolved problem. Here, we describe a nanopore electrochemical sensor to detect ppb-levels of aldehydes arising from lipid degradation in LNP formulations that can be deployed in massively parallel fashion. Specifically, we combine nanopore electrodes with a block copolymer (BCP) membrane capable of hydrophobic gating of analyte transport between the bulk solution and the nanopore volume. By incorporating aldehyde dehydrogenase (ALDH), enzymatic oxidation of aldehydes generates NADH to enable ultrasensitive voltammetric detection with limits-of-detection (LOD) down to 1.2 ppb. Sensor utility was demonstrated by detecting degradation of N-oxidized SM-102, the ionizable lipid in Moderna's SpikeVax™ vaccine, in mRNA-1273 LNP formulation. This work should be of significant use in the pharmaceutical industry, paving the way for automated on-line quality assessments of next-generation therapeutics.

Quantitative analyses of major enzyme activities in postharvest fruit

AbstractThe quantitative determination of major enzyme activities in postharvest fruit is a necessary biochemical experiment to investigate the physiological and metabolic mechanisms of fruit ripeness, senescence, and stress responses. However, it is necessary to choose appropriate methods to stably extract and determine target analytes from complex plant matrices due to the dynamic changes in enzyme activities of postharvest fruit during storage. Here, we outline sensitive and accurate methods for measuring fruit enzyme activities associated with key physiological processes including respiration, reactive oxygen species scavenging, membrane lipid degradation, energy synthesis, cell walls degradation, and disease resistance. This manuscript aims to provide guidance for future research in postharvest biology.

Enhancing shelf life of bell peppers through preharvest fertigation with calcium and potassium thiosulfate: A focus on antioxidant and cell wall degradation enzymes

In this study, the effectiveness of preharvest fertigation using calcium thiosulfate (CaTS) and potassium thiosulfate (KTS) to enhance the shelf life of bell peppers was investigated. The nutrient solution was enriched with KTS (either 0.26 or 0.53 mM of a commercial formulation) or CaTS (0.66 mM of a commercial formulation), and compared to a commercial source-based nutrient solution as a control. Fruit quality attributes and the activity of key enzymes involved in oxidative stress defense, enzymatic browning, cell wall degradation, and membrane lipid degradation were investigated for 30 days of storage. Results showed that CaTS and KTS treatments preserved protein (a secondary energy reserve) and proline content, and enhanced the fruit's PSII activity throughout storage. Despite increased phenylalanine ammonia-lyase activity, CaTS and KTS fertigation did not affect polyphenol oxidase activity. CaTS and KTS fertigation increased the activity of five (out of seven) antioxidant enzymes and impaired the activity of one (out of two) cell wall hydrolytic enzyme. CaTS was the most effective treatment, followed by KTS (0.53 mM), in causing these promotive effects. In conclusion, preharvest application of CaTS (0.66 mM) and KTS (0.53 mM) prolonged the postharvest life and delayed senescence of bell pepper fruits by preserving fruit weight, firmness, and photosynthetic performance, as well as enhancing the antioxidant defense system.

Stability of Liposomal Membrane of Hemoglobin-Vesicles (Artificial Red Cells) for Over Years of Storage Evaluated Using Liquid Chromatography-Mass Spectrometry.

Hemoglobin-Vesicles (Hb-V) are artificial oxygen carriers encapsulating a purified and concentrated Hb solution in liposomes composed of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), cholesterol, 1,5-O-dihexadecyl-N-succinyl-l-glutamate (DHSG), and 1,2-distearoyl-sn-glycero-3-phosphatidylethanolamine-N-poly(ethylene glycol) (PEG5000) (DSPE-PEG). The safety and efficacy of Hb-V have been studied extensively by both preclinical and clinical test methods. Deoxygenation of Hb-V prevents autoxidation of Hb and can extend its shelf life to 2 years at room temperature. However, the lipid components raise concerns about hydrolysis because Hb-V is dispersed in saline. For this study, we attempted to estimate the lipid degradation of long-term stored Hb-V using liquid chromatography-mass spectrometry. Analyses of lipid components extracted from the stored Hb-V showed that the degradation increased depending on the storage temperature. The calculated % remaining of intact lipids of Hb-V were 98.1% after 4 years and 90.4% after 7.2 years at 4 °C, 95.8% after 1 year and 86.7% after 2 years at 25 °C, and 85.6% after 6 months at 40 °C. The main degradation products were lyso-PC and palmitic acid which are hydrolyzed at the ester bond of DPPC. A few hydrolyzed products of DHSG and DSPE-PEG were also detected in Hb-V, but almost no degradation or oxidation products derived from cholesterol could be identified. A shear test of Hb-V at 1500 s-1 showed no significant increase in Hb leakage after storage of 2 years at 25 °C and 6 months at 40 °C. Lipid degradation products including free fatty acids would decrease the pH of the Hb-V dispersion and synergistically facilitate degradation, but it maintained pH 6.5 during 6 years at 4 °C, 2 years at 25 °C, and 3 months at 40 °C because of its high buffering capacity. These results indicate that the storage conditions for Hb-V are appropriate to minimize lipid degradation in the long term.

CsSNF1 kinase plays important roles in fungal development and virulence of the pepper anthracnose fungus Colletotrichum scovillei

AbstractPepper is a vegetable that is used and cultivated worldwide. The quality and production of pepper are reduced by fruit anthracnose disease caused by Colletotrichum scovillei. The highly conserved fungal sucrose non‐fermenting 1 (SNF1)‐related AMPK protein kinases are implicated in the pathogenicity and differentiation of several destructive foliar fungal pathogens; in addition, they affect fungal growth in the presence of glucose at different concentrations and under different environmental stresses. However, the function of SNF1 in C. scovillei is unclear. We functionally characterized SNF1 of C. scovillei using a targeted deletion mutant (ΔCssnf1) generated by homology‐dependent replacement. ΔCssnf1 was severely impaired in mycelial growth under glucose‐limited conditions; multiple defects in conidiation, conidial germination, appressorium formation, lipid degradation and vacuole formation were also observed. Compared to the wild type, ΔCssnf1 caused anthracnose disease of reduced severity on intact and wounded pepper fruits. These defects were recovered in the complemented strain (Cssnf1c). Taken together, our findings implicate CsSNF1 in the growth, development and virulence of C. scovillei.

Endogenous Cellular Metabolite Methylglyoxal Induces DNA-Protein Cross-Links in Living Cells.

Methylglyoxal (MGO) is an electrophilic α-oxoaldehyde generated endogenously through metabolism of carbohydrates and exogenously due to autoxidation of sugars, degradation of lipids, and fermentation during food and drink processing. MGO can react with nucleophilic sites within proteins and DNA to form covalent adducts. MGO-induced advanced glycation end-products such as protein and DNA adducts are thought to be involved in oxidative stress, inflammation, diabetes, cancer, renal failure, and neurodegenerative diseases. Additionally, MGO has been hypothesized to form toxic DNA-protein cross-links (DPC), but the identities of proteins participating in such cross-linking in cells have not been determined. In the present work, we quantified DPC formation in human cells exposed to MGO and identified proteins trapped on DNA upon MGO exposure using mass spectrometry-based proteomics. A total of 265 proteins were found to participate in MGO-derived DPC formation including gene products engaged in telomere organization, nucleosome assembly, and gene expression. In vitro experiments confirmed DPC formation between DNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), as well as histone proteins H3.1 and H4. Collectively, our study provides the first evidence for MGO-mediated DNA-protein cross-linking in living cells, prompting future studies regarding the relevance of these toxic lesions in cancer, diabetes, and other diseases linked to elevated MGO levels.

Conjugates of glycosylated antarctic krill proteins and curcumin: Maintaining the storage quality of salmon fillets coupling with photodynamic inactivation

Antarctic krill proteins (AKP) were successfully glycosylated by inulin driven by ultrasound (300 W, 20 min) and pH shift (11.0) to prepare the conjugates of glycosylated AKP and curcumin. The preservation effects of the conjugates-mediated photodynamic inactivation (PDI) on the storage quality of salmon fillets were investigated. Results showed that the glycosylated AKP (AKP-pH11-U20) exhibited a significantly increased solubility, and a relatively small and loose morphology characterized by SEM, an unfolded and disorganized structure by FTIR and intrinsic fluorescence analysis, and a decreased surface hydrophobicity by bromophenol blue staining, and all these changes contributed to its reduced surface tension. On this basis, the glycosylated AKP efficiently loaded curcumin (323.4 μg/mg) to form the conjugates, and the conjugates-mediated PDI greatly inhibited 99.8% bacterial cells and the migration of water, retarded the change of color and pH, suppressed the lipid oxidation and degradation during 4 °C storage for 7 days, compared to the conventional curcumin-mediated PDI. This study expands the application of new marine protein resources in preserving the quality of aquatic products.

Integrated multiple-omics reveals the regulatory mechanism underlying the effects of homologous Bacillus tequilensis (GCB-3) on growth performance of grass carp (Ctenopharyngodon idellus)

The purpose of this study was to investigate the regulatory mechanism of Bacillus tequilensis (GCB-3) in improving the growth performance of grass carp by microbiome and targeted metabolomics analysis. Fish were fed with different amounts of GCB-3 including B0 (0 CFU/g), B6 (1×106 CFU/g), B7 (1×107 CFU/g), B8 (1×108 CFU/g), and B9 (1×109 CFU/g) for 8 weeks. The results indicate that with the increase of dietary GCB-3 concentration, grass carp showed a significant linear trend in growth performance, and appetite factors showed a significant linear and quadratic trend, the growth performance and appetite-promoting factors in the B8 group reach their maximum value in the B8 group. Microbiome analysis found that Proteobacteria and Actinobacteria were the most abundant phylum in the gut microbiota of grass carp. The functional potential predicted that GCB-3 affected the synthesis and degradation of carbohydrates, amino acids, fatty acids and lipids. It found that GCB-3 also affected the tricarboxylic acid (TCA) cycle VII (acetate-producers) pathway. The results of targeted metabolomics showed a significant linear and quadratic increase in intestinal acetate with GCB-3 concentration and a maximum in the B8 group. The results of the correlation analysis showed a significant positive correlation between Actinobacteria abundance and acetic acid content, and also between TCA cycle VII (acetate-producers) pathway abundance and Actinobacteria abundance and acetic acid content. In B8 group, grass carp showed a faster regulation speed of blood glucose levels after glucose injection, and the GLP-1 content remained stable. The activities of glycolytic enzymes, lipolytic enzymes, protein metabolism enzymes, and their related mRNA expression levels in grass carp showed a linear or quadratic increasing trend. However, the expression levels of gluconeogenic enzymes and fatty acid synthesis enzyme related mRNA showed an opposite trend. In summary, the addition of 1×108 CFU/g GCB-3 in grass carp feed can improve the abundance of dominant bacterial phyla in the intestine, especially Actinobacteria, increase the concentration of intestinal acetate, provide substrate for the TCA cycle VII (acetate-producers) pathway, thereby regulating the nutrient metabolism of grass carp and ultimately promoting growth.

Metabolomics and transcriptomic profiles reveal membrane lipid metabolism being an important factor of sliced taro browning

Cut-surface browning poses a prominent concern when it comes to the storage and consumption of sliced taro. This study evaluated the browning development of sliced taro during cold storage and investigated the underlying browning mechanisms through metabolomics and transcriptomic analyses. Metabolomics analysis revealed a consistent increase in the abundance of metabolites, specifically linolenic acid and its derivatives, as well as hydroperoxides, during the aggravated browning process, suggesting the degradation of membrane lipids, as linolenic acid is a major structural composition of cell membranes. Transcriptomic analysis showed that differentially expressed genes having upregulated expression during the browning process were enriched in the α-Linolenic acid metabolism pathway. Weighted gene co-expression network analysis identified two highly correlated modules with taro slice browning during storage, and genes showing a positive correlation were also enriched in the α-Linolenic acid metabolism pathway. Taken together, these findings indicate that the degradation of membrane lipids during taro browning and the loss of membrane compartmentalization might be key factors contributing to sliced taro browning. Additionally, the upregulation of LOX genes and increased protein activity, along with elevated levels of MDA during browning development, further validate the correlation between membrane lipid peroxidation and taro browning. Overall, this study contributes to our knowledge of the browning mechanisms in fresh-cut foods, providing valuable insights for future research and potential strategies for mitigating browning in sliced taro under cold storage conditions.

Overexpression of GPX2 gene regulates the development of porcine preadipocytes and skeletal muscle cells through MAPK signaling pathway.

Glutathione peroxidase 2 (GPX2) is a selenium-dependent enzyme and protects cells against oxidative damage. Recently, GPX2 has been identified as a candidate gene for backfat and feed efficiency in pigs. However, it is unclear whether GPX2 regulates the development of porcine preadipocytes and skeletal muscle cells. In this study, adenoviral gene transfer was used to overexpress GPX2. Our findings suggest that overexpression of GPX2 gene inhibited proliferation of porcine preadipocytes. And the process is accompanied by the reduction of the p-p38. GPX2 inhibited adipogenic differentiation and promoted lipid degradation, while ERK1/2 was reduced and p-p38 was increased. Proliferation of porcine skeletal muscle cells was induced after GPX2 overexpression, was accompanied by activation in JNK, ERK1/2, and p-p38. Overexpression methods confirmed that GPX2 has a promoting function in myoblastic differentiation. ERK1/2 pathway was activated and p38 was suppressed during the process. This study lays a foundation for the functional study of GPX2 and provides theoretical support for promoting subcutaneous fat reduction and muscle growth.

Comprehensive insights into the effects of acidogenic off-gas utilization on successive biogas production, microbial community structure and metabolite distribution during two-stage anaerobic digestion

Although two-stage anaerobic digestion (TSAD) technology has been investigated, the mechanisms regarding the impact of acidogenic off-gas (AOG) on successive methane production have not been well addressed. In this study, a novel TSAD system was designed. Food waste, as the main substrate, was co-digested with chicken manure and corn straw. The acidogenic gas beyond atmospheric pressure was introduced into the bottom of the methanogenesis reactor through a stainless steel diffuser. Results showed the addition of AOG increased the methane yield from 435.2 to 597.1 mL/g VSin in successive methanogenesis stage, improved by 37.2 %, and increased the energy yield from 9.0 to 11.3 kJ/g VSsubstrate. However, the theoretical contribution of hydrogenotrophic methanogenesis using H2 contained in AOG was only 15.2 % of the increased methane yield. After the addition of AOG, the decreased levels of ammonia nitrogen and butyrate indicate that the stability of the AD system was improved. The electron transfer system and co-enzyme F420 activity were enhanced; however, the decrease in acetate kinase activity indicates aceticlastic methanogenesis may have been weakened. The microbial diversity and species richness were improved by the added AOG. Methanosarcina was more competitive than Methanothermobacter, enhancing the syntrophic effect. The relative abundance of protein degradation bacteria norank_f_Anaerolineaceae and lipid degradation bacteria Syntrophomonas was increased. Metabolite analysis confirmed that the addition of AOG promoted amino acid metabolism, the biosynthesis of other secondary metabolism and lipid metabolism. The improved degradation of recalcitrant organic components (lipids and proteins) in food waste was responsible for the increased methane yield. This study provides an in-depth understanding of the impact of AOG utilization on successive methane production and has practical implications for the treatment of food waste.

Formation of lipid-derived volatile products through lipoxygenase (LOX)- and hydroperoxide lyase (HPL)- mediated pathway in oat, barley and soy bean

The aim of this study was to explore the formation of volatile lipid oxidation products by the lipoxygenase (LOX)-hydroperoxide lyase (HPL)-mediated pathway in oat, barley and soy bean. LOX activity was found only in barley and soy bean samples, but the lipase and HPL activity was detected in all samples. HPL showed particularly high activity with 13-hydroperoxides, while the activity was quite low when using 9-hydroperoxides, especially in the oat and barley. The optimum pH for HPL in different samples was similar, i.e., pH 6–7. In this condition, the volatile compounds formed dramatically with aldehydes and furans as the dominant products. Furthermore, a remarkable enzymatic degradation of lipids occurred during the preparation of food models with highly refined rapeseed oil (RO) and rapeseed oil fatty acid (ROFA) emulsions, where the ROFAs were more prone to oxidation than RO. This study shows the significance of lipid-degrading enzymes in plant-food flavour formation.

Detection and quantification of 4-oxo-2-nonenal in plant-based meat and beef

Oxidative degradation of lipids and proteins in food causes loss of nutrition, produces genotoxic oxidative byproducts, and increases health concerns. 4-Oxo-2-nonenal (4-ONE) is a highly reactive and toxic aldehyde, recently identified as a novel lipid peroxidation byproduct. However, naturally formed 4-ONE and its interaction with proteins in food are still not fully understood. In this study, methods for the detection and quantification of 4-ONE were further developed and used. The developed method successfully detected the formed lipid oxidation products (LOPs) and protein oxidation products (POPs) during cooking, as well as after in-vitro digestion of plant-based (PB) meat and beef. The method used exhibited consistencies of detection of 4-ONE in raw, cooked, and in-vitro digested PB meat and beef samples. Furthermore, the findings of this study show that PB meat had significantly higher levels of POPs but lower levels of aldehydes (4-ONE and malondialdehyde) than beef samples. The results revealed the LC–MS/MS method for detecting and quantifying 4-ONE as a novel lipid oxidation marker was promising. Furthermore, the results indicated that the protein in PB meat may be prone to covalent modification of aldehyde and free radical attacks, thus reducing its proteolysis during digestion. Future studies should further assess the covalent binding ability of aldehydes (especially 4-ONE) in different types of protein, as well as the interaction between lipid and protein oxidation. Processors should develop new strategies to avoid undesirable LOPs and POPs in PB meat and beef, to improve their nutrition and relieve long-term health concerns.

Effect of long chain fatty acids on biogas production and biochemical kinetics in anaerobic bioreactors: a review

Long Chain Fatty Acids (LCFAs) are the primary intermediate byproduct of the lipid (fats, oils, and greases) degradation process, and if they accumulate in high concentrations, they can cause failure or reduce the performance of anaerobic bioreactors due to sludge flotation issues, biochemical kinetics problems for soluble substrates, inhibition of microbial activity, and inefficient biogas recovery. Understanding the biochemical kinetics of anaerobic bioreactors requires taking into account the entire process, including microbe growth, substrate degradation, and product synthesis. Biochemical kinetics of anaerobic treatment is the study of polymer biodegradation rates of insoluble organic matter in wastewater, which is the mechanism of bond breaking and bond formation in biochemical reactions. As a result, biochemical kinetics allow for the design of both desired and undesirable reaction phases. The kinetic parameters acquired are utilized to design, operate, and optimize anaerobic bioreactors for wastewater treatment on a technical scale.

Biotic and abiotic factors acting on community assembly in parallel anaerobic digestion systems from a brewery wastewater treatment plant

ABSTRACT Anaerobic digestion is a complex microbial process that mediates the transformation of organic waste into biogas. The performance and stability of anaerobic digesters relies on the structure and function of the microbial community. In this study, we asked whether the deterministic effect of wastewater composition outweighs the effect of reactor configuration on the structure and dynamics of anaerobic digester archaeal and bacterial communities. Biotic and abiotic factors acting on microbial community assembly in two parallel anaerobic digestion systems, an upflow anaerobic sludge blanket digestor (UASB) and a closed digester tank with a solid recycling system (CDSR), from a brewery WWTP were analysed utilizing 16S rDNA and mcrA amplicon sequencing and genome-centric metagenomics. This study confirmed the deterministic effect of the wastewater composition on bacterial community structure, while the archaeal community composition resulted better explained by organic loading rate (ORL) and volatile free acids (VFA). According to the functions assigned to the differentially abundant metagenome-assembled genomes (MAGs) between reactors, CDSR was enriched in genes related to methanol and methylamines methanogenesis, protein degradation, and sulphate and alcohol utilization. Conversely, the UASB reactor was enriched in genes associated with carbohydrate and lipid degradation, as well as amino acid, fatty acid, and propionate fermentation. By comparing interactions derived from the co-occurrence network with predicted metabolic interactions of the prokaryotic communities in both anaerobic digesters, we conclude that the overall community structure is mainly determined by habitat filtering.

The impacts of different roasting degrees on aroma compounds profiles of moringa seed kernels and its oil

The impacts of different roasting conditions (temperature and duration) on aroma compounds profiles of moringa seed kernels (MSK) and its oil (MSKO) were assessed. We found that medium and dark roasting (roasting temperatures ≥175 °C) induced significant increases in total volatile concentrations in both MSK and MSKO. The generation of volatile compounds during roasting was attributed to the complex thermal reactions, including Maillard reaction, Strecker degradation, caramelisation and lipid thermal oxidation and degradation. Among the detected aroma compounds, newly generated N-containing heterocyclic compounds, O-containing heterocyclic compounds, and sulfur-containing compounds were the primary contributors to the volatile profiles. The sensory results demonstrated that roasting could improve the aroma complexity of MSK and its oil, which may expand their application in the food industry.

Downregulation of HMGCS2 mediated AECIIs lipid metabolic alteration promotes pulmonary fibrosis by activating fibroblasts

BackgroundAbnormal lipid metabolism has recently been reported as a crucial signature of idiopathic pulmonary fibrosis (IPF). However, the origin and biological function of the lipid and possible mechanisms of increased lipid content in the pathogenesis of IPF remains undetermined.MethodsOil-red staining and immunofluorescence analysis were used to detect lipid accumulation in mouse lung fibrosis frozen sections, Bleomycin-treated human type II alveolar epithelial cells (AECIIs) and lung fibroblast. Untargeted Lipid omics analysis was applied to investigate differential lipid species and identified LysoPC was utilized to treat human lung fibroblasts and mice. Microarray and single-cell RNA expression data sets identified lipid metabolism-related differentially expressed genes. Gain of function experiment was used to study the function of 3-hydroxy-3-methylglutaryl-Coa Synthase 2 (HMGCS2) in regulating AECIIs lipid metabolism. Mice with AECII-HMGCS2 high were established by intratracheally delivering HBAAV2/6-SFTPC- HMGCS2 adeno-associated virus. Western blot, Co-immunoprecipitation, immunofluorescence, site-directed mutation and flow cytometry were utilized to investigate the mechanisms of HMGCS2-mediated lipid metabolism in AECIIs.ResultsInjured AECIIs were the primary source of accumulated lipids in response to Bleomycin stimulation. LysoPCs released by injured AECIIs could activate lung fibroblasts, thus promoting the progression of pulmonary fibrosis. Mechanistically, HMGCS2 was decreased explicitly in AECIIs and ectopic expression of HMGCS2 in AECIIs using the AAV system significantly alleviated experimental mouse lung fibrosis progression via modulating lipid degradation in AECIIs through promoting CPT1A and CPT2 expression by interacting with PPARα.ConclusionsThese data unveiled a novel etiological mechanism of HMGCS2-mediated AECII lipid metabolism in the genesis and development of pulmonary fibrosis and provided a novel target for clinical intervention.

Correlations of dynamic changes in lipid and protein of salted large yellow croaker during storage

Protein and lipid are two major components that undergo significant changes during processing of aquatic products. This study focused on the protein oxidation, protein conformational states, lipid oxidation and lipid molecule profiling of salted large yellow croaker during storage, and their correlations were investigated. The degree of oxidation of protein and lipid was time-dependent, leading to an increase in carbonyl content and surface hydrophobicity, a decrease in sulfhydryl groups, and an increase in conjugated diene, peroxide value and thiobarbituric acid reactive substances value. Oxidation caused protein structure denaturation and aggregation during storage. Lipid composition and content changed dynamically, with polyunsaturated phosphatidylcholine (PC) was preferentially oxidized compared to polyunsaturated triacylglycerol. Correlation analysis showed that the degradation of polyunsaturated key differential lipids (PC 18:2_20:5, PC 16:0_22:6, PC 16:0_20:5, etc.) was closely related to the oxidation of protein and lipid. The changes in protein conformation and the peroxidation of polyunsaturated lipids mutually promote each other’s oxidation process.