Cell Membrane Formation Research Articles

Regulation of Milk Fat Synthesis: Key Genes and Microbial Functions

Milk is rich in a variety of essential nutrients, including fats, proteins, and trace elements that are important for human health. In particular, milk fat has an alleviating effect on diseases such as heart disease and diabetes. Fatty acids, the basic units of milk fat, play an important role in many biological reactions in the body, including the involvement of glycerophospholipids and sphingolipids in the formation of cell membranes. However, milk fat synthesis is a complex biological process involving multiple organs and tissues, and how to improve milk fat of dairy cows has been a hot research issue in the industry. There exists a close relationship between milk fat synthesis, genes, and microbial functions, as a result of the organic integration between the different tissues of the cow’s organism and the external environment. This review paper aims (1) to highlight the synthesis and regulation of milk fat by the first and second genomes (gastrointestinal microbial genome) and (2) to discuss the effects of ruminal microorganisms and host metabolites on milk fat synthesis. Through exploring the interactions between the first and second genomes, and discovering the relationship between microbial and host metabolite in the milk fat synthesis pathway, it may become a new direction for future research on the mechanism of milk fat synthesis in dairy cows.

An overview of the cholesterol metabolism and its proinflammatory role in the development of MASLD.

Cholesterol is an essential lipid molecule in mammalian cells. It is not only involved in the formation of cell membranes but also serves as a raw material for the synthesis of bile acids, vitamin D, and steroid hormones. Additionally, it acts as a covalent modifier of proteins and plays a crucial role in numerous life processes. Generally, the metabolic processes of cholesterol absorption, synthesis, conversion, and efflux are strictly regulated. Excessive accumulation of cholesterol in the body is a risk factor for metabolic diseases such as cardiovascular disease, type 2 diabetes, and metabolic dysfunction-associated steatotic liver disease (MASLD). In this review, we first provide an overview of the discovery of cholesterol and the fundamental process of cholesterol metabolism. We then summarize the relationship between dietary cholesterol intake and the risk of developing MASLD, and also the animal models of MASLD specifically established with a cholesterol-containing diet. In the end, the role of cholesterol-induced inflammation in the initiation and development of MASLD is discussed.

Two natural compounds as potential inhibitors against the Helicobacter pylori and Acinetobacter baumannii IspD enzymes

In a vast majority of bacteria, protozoa and plants, the methylerythritol phosphate (MEP) pathway is utilized for the synthesis of isopentenyl diphosphate (IDP) and dimethylallyl diphosphate (DMADP), which are precursors for isoprenoids. Isoprenoids, such as cholesterol and coenzyme Q, play a variety of crucial roles in physiological activities, including cell-membrane formation, protein degradation, cell apoptosis, and transcription regulation. In contrast, humans employ the mevalonate (MVA) pathway for the production of IDP and DMADP, rendering proteins in the MEP pathway appealing targets for antimicrobial agents. This pathway consists of seven consecutive enzymatic reactions, of which 4-diphosphocytidyl-2C-methyl-D-erythritol synthase (IspD) and 2C-methyl-D-erythritol 2,4-cyclodiphosphate synthase (IspF) catalyze the third and fifth steps, respectively. In this study, we characterized the enzymatic activities and protein structures of Helicobacter pylori IspDF and Acinetobacter baumannii IspD. Then, using the direct interaction-based thermal shift assay, we conducted a compound screening of an approved drug library and identified 27 hit compounds potentially binding to AbIspD. Among them, two natural products, rosmarinic acid and tanshinone IIA sodium sulfonate, exhibited inhibitory activities against HpIspDF and AbIspD, by competing with one of the substrates, MEP. Moreover, tanshinone IIA sodium sulfonate also demonstrated certain antibacterial effects against H. pylori. In summary, we identified two IspD inhibitors from approved ingredients, broadening the scope for antibiotic discovery targeting the MEP pathway.

Transcriptomic analysis of cell envelope inhibition by prodigiosin in methicillin-resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a leading threat to public health as it is resistant to most currently available antibiotics. Prodigiosin is a secondary metabolite of microorganisms with broad-spectrum antibacterial activity. This study identified a significant antibacterial effect of prodigiosin against MRSA with a minimum inhibitory concentration as low as 2.5 mg/L. The results of scanning electron microscopy, crystal violet staining, and confocal laser scanning microscopy indicated that prodigiosin inhibited biofilm formation in S. aureus USA300, while also destroying the structure of the cell wall and cell membrane, which was confirmed by transmission electron microscopy. At a prodigiosin concentration of 1.25 mg/L, biofilm formation was inhibited by 76.24%, while 2.5 mg/L prodigiosin significantly reduced the vitality of MRSA cells in the biofilm. Furthermore, the transcriptomic results obtained at 1/8 MIC of prodigiosin indicated that 235and 387 genes of S. aureus USA300 were significantly up- and downregulated, respectively. The downregulated genes were related to two-component systems, including the transcriptional regulator LytS, quorum sensing histidine kinases SrrB, NreA and NreB, peptidoglycan biosynthesis enzymes (MurQ and GlmU), iron-sulfur cluster repair protein ScdA, microbial surface components recognizing adaptive matrix molecules, as well as the key arginine synthesis enzymes ArcC and ArgF. The upregulated genes were mainly related to cell wall biosynthesis, as well as two-component systems including vancomycin resistance-associated regulator, lipoteichoic acid biosynthesis related proteins DltD and DltB, as well as the 9 capsular polysaccharide biosynthesis proteins. This study elucidated the molecular mechanisms through which prodigiosin affects the cell envelope of MRSA from the perspectives of cell wall synthesis, cell membrane and biofilm formation, providing new potential targets for the development of antimicrobials for the treatment of MRSA.

Scopoletin negatively regulates the HOG pathway and exerts antifungal activity against Botrytis cinerea by interfering with infection structures, cell wall, and cell membrane formation

Botrytis cinerea is the causative agent of gray mold, a fungal disease affecting a broad array of food crops and resulting in considerable agricultural and economic losses. Currently, chemical fungicides are the most effective control measure, but this practice is challenged by the growing resistance of pathogens to these fungicides. Scopoletin, a major phytoalexin of tobacco plants, has shown promise as an antifungal treatment. Here, we found that scopoletin was effective against B. cinerea infection of tomato leaves. It inhibited mycelial growth and conidial germination of B. cinerea and disrupted cell wall, cell membrane, and infection structure formation. Transcriptomic analysis identified 3495 differentially expressed genes in response to scopoletin treatment. Interestingly, scopoletin seems to deactivate the HOG pathway of B. cinerea. The growth defect caused by hyperactivation of the pathway, such as iprodione treatment and deletion of BcPtc1, the negative regulator of this pathway, was partly rescued by scopoletin. Moreover, it could downregulate phosphorylation levels of BcSak1 treated with iprodione and NaCl. In addition, the control efficiency of triadimefon was significantly increased by scopoletin. Thus, scopoletin is a promising, naturally derived, and sustainable treatment for gray mold, especially when combined with triadimefon.

Applications of Proteomics in Probiotics Having Anticancer and Chemopreventive Properties.

Proteomics has grown in importance in molecular sciences because it gives vital information on protein identification, expression levels, and alteration. Cancer is one of the world's major causes of death and is the major focus of much research. Cancer risk is determined by hereditary variables as well as the body's immunological condition. Probiotics have increasing medical importance due to their therapeutic influence on the human body in the prevention and treatment of numerous chronic illnesses, including cancer, with no adverse effects. Several anticancer, anti-inflammatory, and chemopreventive probiotics are studied using different proteomic approaches like two-dimensional gel electrophoresis, liquid chromatography-mass spectrometry, and matrix-assisted laser desorption/ionization mass spectrometry. To gain relevant information about probiotic characteristics, data from the proteomic analysis are evaluated and processed using bioinformatics pipelines. Proteomic studies showed the significance of different proteomic approaches in characterization, comparing strains, and determination of oxidative stress of different probiotics. Moreover, proteomic approaches identified different proteins that are involved in glucose metabolism and the formation of cell walls or cell membranes, and the differences in the expression of critical enzymes in the HIF-1 signaling pathway, starch, and sucrose metabolism, and other critical metabolic pathways.

Choline kinase and miR-32-5p: A crucial interaction promoting apoptosis and delaying wound repair in cervical cancer cells

<p>Cervical cancer remains a significant global health challenge, especially in low- and middle-income countries. The elucidation of the molecular pathways underlying the carcinogenesis of cervical cancer is vital to develop targeted therapies. Our study investigates the regulatory effects of miR-32-5p on the choline kinase alpha (<italic>chka</italic>) gene in HeLa cells, a well-established cervical cancer cell line. Choline kinase alpha is recognized for its role in phosphatidylcholine biosynthesis, which is crucial for cell membrane formation, and is implicated in the oncogenic transformation of cells. Utilizing a combination of in silico prediction, luciferase assays, RT-qPCR, and Western blot analyses, we demonstrated that miR-32-5p directly targets the 3′ untranslated region (3′UTR) of <italic>chka</italic> mRNA, leading to a significant downregulation of <italic>chka</italic> expression. Our results demonstrate that miR-32-5p significantly downregulates <italic>chka</italic> at both the mRNA and protein levels, thus leading to decreased cellular proliferation and increased apoptosis. This was further confirmed by a cell cycle analysis, which showed a notable arrest in the G0/G1 phase. Additionally, scratch assays indicated a reduced migratory capacity in miR-32-5p-transfected cells, suggesting the potential anti-metastatic properties of miR-32-5p. These findings highlight the therapeutic potential of miR-32-5p as a biomarker and a target in cervical cancer treatment strategies. By modulating <italic>chka</italic> expression, miR-32-5p could serve as a novel approach to curb the progression and spread of cervical cancer, thus offering a promising avenue for future research and clinical applications. This study contributes to the growing understanding of miRNA-mediated gene regulation in cancer biology and underscores the importance of targeted genetic research in the development of cancer therapeutics.</p>

Inhibitors of 3-Hydroxy-3-methylglutaryl Coenzyme A Reductase Decrease the Growth, Ergosterol Synthesis and Generation of petite Mutants in Candida glabrata and Candida albicans

Candida glabrata and Candida albicans, the most frequently isolated candidiasis species in the world, have developed mechanisms of resistance to treatment with azoles. Among the clinically used antifungal drugs are statins and other compounds that inhibit 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), resulting in decreased growth and ergosterol levels in yeasts. Ergosterol is a key element for the formation of the yeast cell membrane. However, statins often cause DNA damage to yeast cells, facilitating mutation and drug resistance. The aim of the current contribution was to synthesize seven series of compounds as inhibitors of the HMGR enzyme of Candida ssp., and to evaluate their effect on cellular growth, ergosterol synthesis and generation of petite mutants of C. glabrata and C. albicans. Compared to the reference drugs (fluconazole and simvastatin), some HMGR inhibitors caused lower growth and ergosterol synthesis in the yeast species and generated fewer petite mutants. Moreover, heterologous expression was achieved in Pichia pastoris, and compounds 1a, 1b, 6g and 7a inhibited the activity of recombinant CgHMGR and showed better binding energy values than for α-asarone and simvastatin. Thus, we believe these are good candidates for future antifungal drug development.

Fatty Acid Metabolism is Associated with Immune Microenvironment Variation in Bladder Cancer, and FN1 May Mediate PD-L1 Expression through the IL6-STAT3 Pathway

Background: Fatty acid metabolism plays an important role in many biological activities, such as cell membrane formation, energy storage, and signal molecule generation in tumorigenesis. Lipid metabolism affects the progression and treatment of Bladder Cancer (BLCA). Therefore, it is imperative to explore the function and prognostic value of lipid metabolism-related genes in BLCA patients. Methods: In this study, we collected gene expression profiles and clinical information in The Cancer Genome Map (TCGA) database and two independent Group on Earth Observations (GEO) datasets. Gene interaction information was obtained from ENCORI database. Based on these databases, we analyzed the expression patterns of genes and proteins involved in fatty acid metabolism and their matching clinicopathological features. Further, the gene for fatty acid metabolism, FN1, was screened for cellular function science experiments to validate our findings. Results: Analysis in the TCGA database identified 310 fatty acid metabolism-related mRNAs, 91 of which were differentially expressed in BLCA patients. Based on the correlation of Differentially Expressed Genes (DGEs) with patient characteristics, we developed a clinical prognostic correlation model and validated the accuracy of the model based on information from the GEO database. Survival analysis and clinical correlation analysis showed that elevated FN1 levels were highly correlated with shorter survival, higher staging, higher grading, and lower infiltration of immune cells in BLCA. Furthermore, we experimentally verified that FN1 can activate the IL6 -JAK - STAT3 pathway and further promote PD-L1 expression, which would serve as a potential factor for immune escape in bladder cancer. Finally, our experimental results were consistent with the bioinformatics analysis.

R/K selection of GC content in prokaryotes.

The guanine/cytosine (GC) content of prokaryotic genomes is species-specific, taking values from 16% to 77%. This diversity of selection for GC content remains contentious. We analyse the correlations between GC content and a range of phenotypic and genotypic data in thousands of prokaryotes. GC content integrates well with these traits into r/K selection theory when phenotypic plasticity is considered. High GC-content prokaryotes are r-strategists with cheaper descendants thanks to a lower average amino acid metabolic cost, colonize unstable environments thanks to flagella and a bacillus form and are generalists in terms of resource opportunism and their defence mechanisms. Low GC content prokaryotes are K-strategists specialized for stable environments that maintain homeostasis via a high-cost outer cell membrane and endospore formation as a response to nutrient deprivation, and attain a higher nutrient-to-biomass yield. The lower proteome cost of high GC content prokaryotes is driven by the association between GC-rich codons and cheaper amino acids in the genetic code, while the correlation between GC content and genome size may be partly due to functional diversity driven by r/K selection. In all, molecular diversity in the GC content of prokaryotes may be a consequence of ecological r/K selection.

Regulation of Phosphate Transporters and Novel Regulator of Phosphate Metabolism

Phosphorus is essential for all living organisms. It plays an important role in maintaining biological functions, such as energy metabolism, cell membrane formation, and bone mineralization. Various factors in the intestine, kidneys, and bones regulate the homeostasis of the inorganic phosphate (Pi) concentration in the body. X-linked hypophosphatemia (XLH), the most common form of hereditary hypophosphatemic rickets, is characterized by an impaired mineralization of the bone matrix, hypertrophic chondrocytes with hypophosphatemia, and active vitamin D resistance in childhood. Phosphate-regulating gene with homologies to endopeptidases on the X chromosome was recognized as the responsible gene for XLH. XLH is classified as fibroblast growth factor 23 (FGF23)-related hypophosphatemic rickets. The enhanced FGF23 stimulates renal phosphate wasting by downregulating sodium-dependent Pi cotransporters, NaPi2a and NaPi2c proteins, in the proximal tubules. Recently, transmembrane protein (Tmem) 174 has been identified as a novel regulator of phosphate transporters. This review introduces the role of Tmem174 in the Pi homeostasis in the body.

Interactions between phenanthrene and polystyrene micro/nano plastics: Implications for rice (Oryza sativa L.) toxicity

Micro/nano plastics (MPs/NPs) are widely distributed and are one of the global pollutants of current concern. Micro/nano plastics can adsorb a variety of persistent organic pollutants, and different particle sizes and surface charges affect the biological effects of MPs/NPs. Therefore, how the compound pollution of MPs/NPs with different particle sizes and organic pollutants produces toxic effects on plants needs to be further studied. We investigated the toxic effects of phenanthrene (Phe) and amino-modified PS (PS–NH2) with two particle sizes (50 nm, 5 μm) on rice. The stress mechanism of PS-NH2 was different between the two particle sizes. Moreover, 50 nm PS-NH2 inhibited stomatal conductance and transpiration rate, reduced photosynthetic rate, significantly enriched GO functions such as “DNA repair” and “DNA double-strand break,” and caused severe DNA damage in rice. Notably, 5 μm PS-NH2 affected the gene expression of “photosynthetic lighting” and “photosynthetic antenna protein” in rice, decreased chlorophyll content, and inhibited rice growth. The toxicity of 50 nm PS-NH2 was stronger. In addition, we found that Phe reduced the toxicity of PS-NH2 with different particle sizes, and the relief effect of 50 nm PS-NH2+Phe was more evident. Further, 50 nm PS-NH2+Phe alleviated the toxicity by stimulating the activities of antioxidant enzymes, reducing oxidative damage to chloroplasts, and inhibiting photosynthesis. However, 5 μm PS-NH2+Phe can reduce the stress by reducing the degree of membrane lipid peroxidation, activating metabolic pathways related to the cell wall and cell membrane formation, and plant antitoxin biosynthesis. The results contribute to the understanding of the mechanism of toxicity of MPs/NPs and polycyclic aromatic hydrocarbons (PAHs) to crops.

Antibacterial mechanism of vanillin against Escherichia coli O157: H7

Vanillin, a plant-derived antimicrobial volatile substance, has potential microbial control applications in the food industry. However, the effect of vanillin on the food-borne pathogen Escherichia coli (E. coli) O157:H7 has not been well studied. This study aims to explore the antibacterial mechanism of vanillin against E. coli O157:H7. The minimum inhibitory concentration (MIC) and antibacterial effect of vanillin were determined by microdilution. Scanning electron microscopy (SEM) was used to observe the damage of vanillin to the cell membrane, while cell membrane potential and the leakage of nucleic acid protein were measured to explore the effect of vanillin on the membrane system. Confocal laser scanning and intracellular adenosine triphosphate (ATP) concentration determination were utilized to investigate the effects of vanillin on the energy, life, and death of E. coli. Finally, transcriptome sequencing was conducted to investigate the gene expression differences induced by vanillin treatment. The results showed that vanillin treatment effectively controlled E. coli O157:H7 with an MIC of 2 mg/mL. After treatment, damage to the membrane system, depolarization of the membrane, and leakage of nucleic acid and protein were observed. Meanwhile, vanillin treatment caused decreased ATP content and cell death. Transcriptome analysis showed that vanillin treatment significantly affected the expression of genes involved in cell membrane formation, tricarboxylic acid (TCA) cycling pathway, and oxidative phosphorylation pathway in E. coli O157:H7. In conclusion, membrane damage and energy metabolism disruption are important mechanisms of vanillin's inhibitory effect on E. coli O157:H7. This study provides new insights into the molecular reaction mechanism of vanillin against E. coli O157:H7, highlighting its potential as an antibacterial substance for preventing E. coli contamination in the food industry.

A strain of Vibrio alginolyticus isolated from Azumapecten farreri and its pathogenic mechanism using CRISPR-Cas9 technology.

Scallops have become an important aquaculture species in China because they contain high-quality protein, and scallops are important health food that combines multiple effects and high economic benefits. However, scallop aquaculture is perennially threatened by various pathogenic Vibrio species, leading to great economic losses. We obtained a strain of pathogenic bacteria, identified as Vibrio alginolyticus, from the diseased Azumapecten farreri in the scallop farming area of Huangdao District in 2018, and V. alginolyticus is one of the major shellfish pathogens. We showed that V. alginolyticus was isolated and identified as a pathogen in A. farreri for the first time. In this study, we evaluated its morphology and performed a phylogenetic analysis based on 16S rRNA gene sequencing. In addition, we performed a preliminary analysis of its pathogenic mechanisms. The Hfq protein in V. alginolyticus is an important RNA-binding protein in the quorum-sensing system that not only affects the sensitivity of Vibrio to environmental stress but also regulates a variety of functions, such as cell membrane formation, motility, and virulence towards the host. However, its effect on the pathogenesis of V. alginolyticus to A. farreri is unclear. To further investigate the pathogenic mechanism of the Hfq protein in V. alginolyticus to A. farreri, we used the CRISPR-Cas9 system to target and deplete the hfq gene fragment in V. alginolyticus and obtained the mutant strain V. ΔHfq-. We found that the peripheral flagellum of the mutant strain was lost, which reduced the motility of V. alginolyticus. Therefore, the deletion of target genes by the CRISPR/Cas9 genome editing system confirmed that the Hfq protein played a key role in reducing the ability of V. alginolyticus to infect A. farreri. In conclusion, our current findings provided valuable insights into the healthy culture of scallops.

Antimicrobial Activity the Essential Oil from Croton pluriglandulosus Carn. Leaves against Microorganisms of Clinical Interest.

Multiresistant pathogens pose a serious threat to human health. The genus Candida is one class of human pathogenic yeasts responsible for infections affecting healthy and immunocompromised patients. In this context, plant essential oils emerged as a future natural alternative to control the diseases caused by these pathogens. Based on that, the present study aimed to evaluate the antimicrobial potential of essential oil from C. pluriglandulosus and understand the mechanism of action. Here, it highlighted antimicrobial activity and the mechanisms of action of the essential oil extracted from C. pluriglandulosus Carn.-Torres & Riina (CpEO) leaves on human pathogenic microorganisms in planktonic and biofilm lifestyles. In addition, for the first time, the oil composition was revealed by GC-MS analysis and the toxicity to human red blood cells (HRBC). Twenty-six chemical compounds were identified in CpEO, elemicin, bicyclogermacrene, caryophyllene, brevifolin, and 2,4,6-trimethoxy-styrene. Through hemolytic assay, it was shown that CpEO has no toxicity to human RBCs. At the concentration of 50 μg mL-1, CpEO did not show great antibacterial potential. However, promising data were found for C. krusei and C. parapsilosis inhibiting by 89.3% and 80.7% of planktonic cell growth and 83.5% and 77.9% the biofilm formation, respectively. Furthermore, the mechanisms of action CpEO were elucidated by fluorescence. Scanning electron microscopy revealed damage to the cell membrane and pore formation, ROS overproduction, and induction of apoptosis in candida cells. Our results reinforce the potential of CpEO as an effective alternative molecule of pharmaceutical interest.

Antimicrobial Effect of Ocimum gratissimum L. Essential Oil on Shewanella putrefaciens: Insights Based on the Cell Membrane and External Structure.

The main objective of this study was to assess the in vitro antibacterial effectiveness of Ocimum gratissimum L. essential oil (OGEO) against Shewanella putrefaciens. The minimum inhibitory concentration and minimum bactericidal concentration of OGEO acting on S. putrefaciens were both 0.1% and OGEO could inhibit the growth of S. putrefaciens in a dose-dependent manner. The restraint of the biofilm growth of S. putrefaciens was found in the crystal violet attachment assay and confocal laser scanning microscopy. The disruption of cell membranes and exudation of contents in S. putrefaciens with OGEO treatment were observed by scanning electron microscopy, hemolysis and ATPase activity. The results demonstrated that OGEO had a positive inhibitory effect on the growth of S. putrefaciens, which primarily developed its antibacterial function against S. putrefaciens by disrupting the formation of biofilms and cell membranes. This study could provide a new method of inhibiting the spoilage of food in which the dominant spoilage bacteria are S. putrefaciens.

Emerging roles of host and microbial bioactive lipids in inflammatory bowel diseases.

The intestinal tract harbors diverse microorganisms, host- and microbiota-derived metabolites, and potentially harmful dietary antigens. The epithelial barrier separates the mucosa, where diverse immune cells exist, from the lumen to avoid excessive immune reactions against microbes and dietary antigens. Inflammatory bowel disease (IBD), such as ulcerative colitis and Crohn's disease, is characterized by a chronic and relapsing disorder of the gastrointestinal tract. Although the precise etiology of IBD is still largely unknown, accumulating evidence suggests that IBD is multifactorial, involving host genetics and microbiota. Alterations in the metabolomic profiles and microbial community are features of IBD. Advances in mass-spectrometry-based lipidomic technologies enable identification of changes in the composition of intestinal lipid species in IBD. Because lipids have a wide range of functions, including signal transduction and cell membrane formation, dysregulation of lipid metabolism drastically affects the physiology of the host and microorganisms. Therefore, a better understanding of the intimate interactions of intestinal lipids with host cells that are implicated in the pathogenesis of intestinal inflammation might aid in the identification of novel biomarkers and therapeutic targets for IBD. This review summarizes the current knowledge on the mechanisms by which host and microbial lipids control and maintain intestinal health and diseases. This article is protected by copyright. All rights reserved.

Association of serum lipid profile ratios with disease severity and in-hospital outcome of patients with COVID-19

Background: The association of lipid ratios with the severity and hospital outcome of COVID-19 patients has been a topic of interest in the medical community. Lipids, particularly cholesterol and triglycerides, play a significant role in the human body, including cell membrane formation, energy storage and the regulation of various biological processes. The aim of this study was to assess the association of lipid ratio with the severity and hospital outcome of COVID-19 patients. Methods: This observational study was carried out in COVID unit of Bangabandhu Sheikh Mujib Medical University (BSMMU) among hospital admitted COVID patients from March 2021 to February 2022. Results: Total patients were 138 including 65.9% males. Majority (27.5%) were in the 41-50 age group. One third (31.2%) were clinically severe COVID-19. There was overall significant difference of lipid ratio in different severity except LDL/HDL group (p>0.05). The correlation coefficients of disease severity with TC/HDL (rs)=.338, LDL/HDL (rs)=.289 and TG/HDL (rs)=.444. There was significant correlation between disease severity and lipid ratio having moderate correlation with TG/HDL and weak correlation with TC/HDL and LDL/HDL. The correlation coefficients between TC/HDL and hospital outcomes of use/development of rebreather (rs)=.217, HFNC (rs)=.282, ARDS (rs)=.307, arrhythmia (rs) = .201, HDU/ICU (rs)=.200 and ventilator (rs)=.072. There was significant correlation between TC/HDL and development of ARDS, use of rebreather and HFNC and no other significant correlation with other outcomes. The correlation coefficients between LDL/HDL and hospital outcomes of use/development of rebreather (rs)=.211, HFNC (rs)=.132, ARDS (rs)=.286, arrhythmia (rs)=.106, HDU/ICU (rs)=.064 and ventilator (rs)=.028. There was no significant correlation between LDL/HDL and hospital outcomes. The correlation coefficients between TG/HDL and hospital outcomes of use/development of rebreather (rs)=.427, HFNC (rs)=.271, ARDS (rs)=.502, arrhythmia (rs)=.088, HDU/ICU (rs)=.158 and ventilator (rs) =.041. There was significant correlation between TG/HDL and development of ARDS, also with use of rebreather and HFNC. No other significant correlation found with other outcomes. Conclusion: From the findings of this current study, in can be concluded that lipid ratios are associated with disease severity and hospital outcome of COVID-19 patients. There was moderate association between disease severity and TG/HDL and weak association with TC/HDL and LDL/HDL. There was also association between TC/HDL and the development of ARDS and the use of HFNC and between TG/HDL and the development of ARDS, the use of rebreather, and HFNC. BIRDEM Med J 2023; 13(2): 98-102

Determination of 39 fatty acids in liver of rats by gas chromatography-mass spectrometry

Fatty acids not only form phospholipids that contribute to the formation of cell membranes but also participate in many metabolic activities, such as energy storage and cell signal transduction. The liver plays a key role in the synthesis and metabolism of fatty acids. The composition and contents of fatty acids in the liver are closely related to body health. Most fatty acid-detection methods require a large sample size and can detect only a small number of fatty acids. Therefore, a sensitive and efficient method to determine fatty acids in the liver is urgently required. Herein, a method based on gas chromatography-mass spectrometry (GC-MS) was established for the simultaneous determination of 39 fatty acids in 1.1 mg of liver tissue. Different extraction methods and derivatization conditions were compared to develop an optimal sample-treatment method. The performance of two different columns in separating the target fatty acids were also compared. A total of 10 mg of liver was added to 450 μL of normal saline and ground at -35 ℃ to obtain a homogenate. Next, 50 μL of the homogenate (equivalent to 1.1 mg of liver) was added with 750 μL of chloroform-methanol (1∶2, v/v) to extract total fatty acids. The fatty acid extracts were dried under nitrogen, and then derivatized at 100 ℃ for 90 min after being added with methanol containing 5% sulfuric acid. The fatty acid methyl esters were extracted with hexane and then separated on an SP-2560 capillary column (100 m×0.25 mm×0.2 μm; Supelco, USA) via GC-MS. The results revealed that all 39 fatty acid methyl esters detected had good linearities in the certain mass concentration ranges with correlation coefficients (R2) greater than 0.9940. The limits of detection (LOD) and quantification (LOQ) of these methyl esters in the liver were 2-272 ng/mg and 7-906 ng/mg, respectively. The accuracy and precision of the method were evaluated by spiking the liver homogenate with tridecylic acid and eicosanoic acid at low (0.09 μg/mg), moderate (0.90 μg/mg), and high (5.40 μg/mg) concentration levels. The recoveries ranged from 82.4% to 101.0% with an intraday relative standard deviations (RSDs) (n=5) of 3.2%-12.0% and interday RSDs (n=3) of 5.4%-13.4%. The method was successfully applied to detect fatty acids in the livers of four healthy male Sprague-Dawley (SD) rats and four male SD rats with abnormal liver function induced by perfluorooctane sulfonate (PFOS). PFOS is a persistent organic pollutant. Twenty-six fatty acids were detected in the livers of both groups. Among the fatty acids investigated, pentadecanoic acid (C15∶0), γ-linolenic acid (C18∶3n6), and elaidic acid (C18∶1n9t) cannot be detected by the methods reported in the literature. By contrast, the method developed in this study could separate the isomers of oleic acid (elaidic acid, C18∶1n9t; oleic acid, C18∶1n9c) and linolenic acid (linolelaidic acid, C18∶2n6t; linoleic acid, C18∶2n6c). In conclusion, the developed method is simple and can detect a large number of fatty acids using small sample amounts and few reagents. More importantly, it could successfully separate fatty acid isomers. These findings indicate that the developed method is suitable for the detection of fatty acid composition and contents in the liver in clinical and experimental research.

Protein interactomes for plant lipid biosynthesis and their biotechnological applications.

Plant lipids have essential biological roles in plant development and stress responses through their functions in cell membrane formation, energy storage and signalling. Vegetable oil, which is composed mainly of the storage lipid triacylglycerol, also has important applications in food, biofuel and oleochemical industries. Lipid biosynthesis occurs in multiple subcellular compartments and involves the coordinated action of various pathways. Although biochemical and molecular biology research over the last few decades has identified many proteins associated with lipid metabolism, our current understanding of the dynamic protein interactomes involved in lipid biosynthesis, modification and channelling is limited. This review examines advances in the identification and characterization of protein interactomes involved in plant lipid biosynthesis, with a focus on protein complexes consisting of different subunits for sequential reactions such as those in fatty acid biosynthesis and modification, as well as transient or dynamic interactomes formed from enzymes in cooperative pathways such as assemblies of membrane-bound enzymes for triacylglycerol biosynthesis. We also showcase a selection of representative protein interactome structures predicted using AlphaFold2, and discuss current and prospective strategies involving the use of interactome knowledge in plant lipid biotechnology. Finally, unresolved questions in this research area and possible approaches to address them are also discussed.