Quantification Of Fatty Acids Research Articles

Fat extraction methodology can affect the food fatty acid profile: improvement of a protocol more environmentally friendly

Fatty acids (FAs) are biochemical components of food, essential for human health due to their numerous biological functions. However, many of then if consumed in excess can trigger disfunctions/illness. Therefore, analytical methods, such as gas chromatography (GC-FID) are essential for the accurate identification and quantification of FAs, playing an important role in food safety and quality assessment. The aim of this study was to compare two FAs extraction protocols and to adapt, optimize, and validate a new one for food FAs extraction, considering the principles of green chemistry (sustainability and environmentally friendly). The research included four main steps: (A) comparison of two FAs extraction protocols for Gas Chromatography Flame Ionization Detector (GC-FID) analysis; (B) adaptation and optimization of four new extraction protocols; (C) verification of the most promising protocol; and (D) evaluation of the greenness profile. Comparatively, the PE2 protocol detected a greater diversity of FAs and a wider range of medium/long chain saturated fatty acids (SFA), but with divergences from values declared on food labels, mainly in the quantification of unsaturated fatty acids (UFA), unlike PE1, which presented lipid fraction values closer to nutritional information and greater extraction of short/medium chain FAs. The improved protocol, PE6, proved to be more efficient in extracting short, medium, and long chain FAs, as well as having SFA and UFA values closer to nutritional values in different food matrices. Furthermore, PE6 also showed improvements in relation to green chemistry aspects, presenting a higher greenness profile score, which indicates that it is more sustainable and generates less environmental impact. These results underscore the importance and relevance of continuing to develop analytical methods for food that are both effective and environmentally responsible.

GC-MS quantification of fecal short-chain fatty acids and spectrophotometric detection of indole: Do rectal swabs produce comparable results as stool samples? - A pilot study.

The exploration of the gut microbiome and related metabolites holds an exciting future in health science. The challenges associated with fecal sample testing are proper sample collection, sterile transportation, optimal transport conditions, and processing as all these factors could potentially change the microbiome composition, further exacerbated by the patient's customary discomfort regarding feces samples. The study aimed to compare the usage of rectal swabs and stool samples for short-chain fatty acid estimation using gas chromatography-mass spectrometry (GC-MS) and indole estimation using spectrophotometry. From May 2022 to June 2022, three women were recruited from the Department of Obstetrics and Gynecology (OBG) in a secondary care hospital in coastal Karnataka. During their clinical visit, a rectal swab was collected, and the stool sample was transported to the hospital from the patient's home in sterile containers provided. After the extraction, short-chain fatty acids (acetate, propionate, and butyrate) were quantified using GC-MS. The fecal indole concentration was determined using a hydroxylamine-based assay. The GC-MS analysis failed to detect the concentrations of short-chain fatty acids in rectal swab samples. Indole concentrations in stool and swab samples were significantly different. The study's findings do not support the use of rectal swabs to analyze gut metabolites.

Therapeutic potential of compound extract from Dracocephalum Rupestre Hance and Berberidis Radix against Salmonella-induced lamb diarrhea

Lamb diarrhea is primarily induced by bacterial infections, causing great economic and health challenges. Traditional antibiotic treatments raise concerns over drug resistance and environmental contamination. We explored the therapeutic potential of a compound extract from Dracocephalum rupestre Hance and Berberidis Radix against Salmonella-induced diarrhea in lamb. Twenty-five five-week-old Kunming mice (20 ± 5 g) were used. A controlled laboratory experiment, combing histological examinations, serum cytokine level analysis, gut microbiota composition analysis, and short-chain fatty acid quantification were conducted. Results demonstrated significant reparative effects on intestinal mucosal damage of the compound. Compound treatment notably reduced serum levels of inflammatory cytokines (IL-6, IL-8, sigA, and TNF-α), indicating an anti-inflammatory effect. Gene expression analysis of mucosal repair markers (PCNA, TGF, and EGFR) confirmed the positive impacts on intestinal recovery processes after treatment. Microbiota analysis revealed concentration-dependent alterations in gut microbial composition, with a notable increase in beneficial bacterial genera such as Muribaculum and Prevotella, suggesting the role of the compound in promoting gut health. Additionally, short-chain fatty acid analysis indicated an increase in beneficial acids, which are critical for the gut and overall health. This investigation highlights the potential therapeutic benefits of Dracocephalum rupestre Hance combining Berberidis Radix in lamb with Salmonella-induced diarrhea.

Associations of the rs12504538 and rs6824447 Polymorphisms of the Elovl6 Gene with Estimated Elongase and Desaturase Activity and Fatty Acid Concentrations in Mexican Women with Gestational Diabetes Mellitus

Background: This study aimed to investigate the possible associations of the rs12504538 and rs6824447 polymorphisms of the Elovl6 gene with estimated elongase and desaturase activity and saturated fatty acid concentrations in Mexican women with Gestational Diabetes Mellitus (GDM). Methods: We recruited 172 women in the second and third trimesters of pregnancy who had undergone an oral glucose tolerance test, including 66 who had diagnosed with GDM according to the 2016 ADA criteria and 106 who had normal glucose tolerance test results, from the General Hospital and Health Centers of the Health Ministry of Guanajuato, México. Participants were matched by gestational week. Data on age, gestational week and anthropometric characteristics were collected. Blood samples were drawn after an overnight fast for the measurement of serum glucose, lipid, NEFA, serum fatty acid and insulin levels; SNP genotyping and quantification of fatty acids was performed and elongase and desaturase activity was estimated. Findings: With the exception of HDL-cholesterol, all variables, including NEFA levels, were significantly greater in the GDM group than in the non-GDM group. The rs6824447 polymorphism of the Elovl6 gene, an age >25 years and HOMA-IR levels were associated with the development of GDM (OR=5.1, 95% CI 1.56-17.1, p=0.006; OR=4.89, 95% CI 1.65-14.4, p=0.003; OR=34.1 95% CI 8.7-133, p<0.000001, respectively). Palmitic acid (OR=1.08; 95% CI: 1.06-1.11; p<0.001) and oleic acid (OR=1.17; 95% CI: 1.14-1.2; p<0.001) concentrations were also associated with the development of GDM. No differences in the estimated elongase and desaturase activity among the non-GDM and GDM groups were found and only the activity of Δ9D SCD18 desaturase was marginally high in GDM patients (p=0.050). Conclusion: The rs6824447Elovl6 polymorphism is associated with the development of GDM, as are high serum palmitic acid, oleic acid and stearic acid concentrations and estimated Δ9D SCD18 desaturase activity.

Synbiotic supplementation may globally improve non-motor symptoms in patients with stable Parkinson’s disease: results from an open label single-arm study

Gut microbiota changes and brain-gut-axis (BGA) dysregulation are common in people with Parkinson’s Disease (PD). Probiotics and prebiotics are emerging as a potential therapeutic approach for PD patients. The aim of this paper was to assess the neurological and gastroenterological effects in PD patients with constipation after the administration of a synbiotic product, with a focus on behavioral and cognitive symptoms. We enrolled patients with stable PD who met diagnostic criteria for functional constipation and/or irritable bowel syndrome with constipation according to Rome IV Criteria. Patients received a synbiotic treatment (Enterolactis Duo, containing the probiotic strain Lacticaseibacillus paracasei DG and the prebiotic fiber inulin) for 12 weeks. A neurological and a gastroenterological evaluation were collected before and after the treatment. In addition, 16S rRNA gene profiling and short chain fatty acid quantification were performed to characterize the microbial ecosystem of fecal samples collected before (n = 22) and after (n = 9) the synbiotic administration. 30 patients were consecutively enrolled. After treatment, patients performed better in MDS-UPDRS part 1 (p = 0.000), SCOPA-AUT (p = 0.001), TAS-20 (p = 0.014), HAM-D (p = 0.026), DIFt (p = 0.003), PAS-A (p = 0.048). Gastroenterological evaluations showed improvements in PAC-SYM score (p < 0.001), number of complete bowel movement (p < 0.001) and BSFS (p < 0.001). After the synbiotic administration, we observed a significant increase in the abundance of the order Oscillospirales, as well as the Oscillospiraceae family and the species Faecalibacterium prausnitzii within this order in fecal samples. Synbiotic treatment demonstrates potential efficacy in ameliorating non-motor features in PD patients.

An economically viable stable isotope-enhanced multiple reaction monitoring method for total fatty acid analysis in a mouse model of non-alcoholic fatty liver disease

The complex pathological mechanisms of non-alcoholic fatty liver disease (NAFLD) are closely related to dysregulated lipid metabolism, and the therapeutic effects of the traditional Chinese medicine Zexie-Baizhu Decoction (AA) on NAFLD have been gaining increasing attention. However, research into altered lipid metabolism, especially fatty acids, in NAFLD and the intervention of AA faces technical challenges, especially in the precise quantitative analysis of fatty acids in biological samples. The high complexity of biological matrices, particularly after drug intervention, greatly increases the difficulty of detection. Therefore, this study innovatively developed a simple and economical stable isotope derivatization technique by synthesizing d6N,N-dimethylethylenediamine (d6-DMED) in the laboratory, establishing a simple and economical method for fatty acid quantification. This method employs a chemical reaction under low-temperature conditions to ensure the efficient synthesis of d6-DMED. Using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry technique (UHPLC-MS/MS), combined with optimized chromatographic separation conditions and dynamic multiple reaction monitoring mode, the study established a highly sensitive detection method for 35 fatty acid derivatives. Methodological evaluation showed that the limits of quantification ranged from 0.002 to 0.060 μM, with high linearity of R² > 0.995. Additionally, the relative recovery rates were between 93.14% and 106.63%. To further demonstrate the feasibility of this method for fatty acid quantification, it was applied to measure fatty acids in multiple tissues in a mouse NAFLD model, as well as the effects of AA intervention on fatty acid metabolism. This rapid, simple, and cost-effective detection method not only enhances the understanding of NAFLD mechanisms but also provides a new strategy for evaluating the biological complex system after drug intervention.

Making OzID go FFASTer: Combining stable-isotope tagging with ozone-induced dissociation to uncover changes in fatty acid unsaturation within neurosecretory cells

Despite lipids providing a major contribution to the mass of the brain, at the molecular level, the brain lipidome remains incompletely described and the functions of many of the species identified to date are unknown. Recently, unusual unsaturated fatty acids –with carbon-carbon double bonds in positions not predicted by canonical lipid metabolism– were identified in the mouse brain. The extent to which these unusual lipids contribute to neurochemical processes presents a significant gap in knowledge that is challenging to close due to technological impediments to the detection and tracing of such novel fatty acids. Here we deploy state-of-the-art mass spectrometric methods coupled with diagnostic ion-molecule reactions in the form of ozone-induced dissociation to enable detection and relative quantification of unsaturated fatty acids in an in vitro brain model based on the PC12 cell line. This approach revealed the presence of abundant populations of non-canonical fatty acids, including FA 16:1n-10,cis (sapienic acid) and FA 18:1n-10,cis (dihomosapienic acid), in extracts from PC-12 cell lines. Neither of these fatty acids have previously been reported in neurons or neurosecretory-like cells, with both demonstrated to increase between 2 and 4-fold in relative abundance upon active stimulation.

Gut microbiome and function are altered for individuals living in high fluoride concentration areas in Pakistan

BackgroundEndemic fluorosis refers to the condition when individuals are exposed to excessive amounts of fluoride ion due to living in a region characterized by elevated levels of fluorine in the drinking water, food, and/or air. In Pakistan, a substantial proportion of the population is thereby affected, posing a public health concern. ObjectivesAssessing how the gut microbiota and its metabolic profiles are impacted by chronic exposure to fluoride in drinking water (that caused Dental Fluorosis) as well as to perceive how this microbiota is connected to adverse health outcomes prevailing with fluoride exposure. MethodsDrinking water (n=27) and biological samples (n=100) of blood, urine and feces were collected from 70 high fluoride exposed (with Dental Fluorosis) and 30 healthy control (without Dental Fluorosis) subjects. Water and urinary fluoride concentrations were determined. Serum/plasma biochemical testing was performed. Fecal DNA extraction, 16S rRNA analysis of microbial taxa, their predicted metabolic function and fecal short chain fatty acids (SCFAs) quantification were carried out. ResultsThe study revealed that microbiota taxonomic shifts and their metabolic characterization had been linked to certain host clinical parameters under the chronic fluoride exposure. Some sets of genera showed strong specificity to water and urine fluoride concentrations, Relative Fat Mass index and SCFAs. The SCFAs response in fluoride-exposed samples was observed to be correlated with bacterial taxa that could contribute to adverse health effects. ConclusionsMicrobial dysbiosis as a result of endemic fluorosis exhibits a structure that is associated with risk of metabolic deregulation and is implicated in various diseases. Our results may form the development of novel interventions and may have utility in diagnosis and monitoring.

Metabolic profiling of pea (Pisum sativum) cultivars in changing environments: Implications for nutritional quality in animal feed

Field pea seeds have long been recognized as valuable feed ingredients for animal diets, due to their high-quality protein and starch digestibility. However, the chemical composition of pea cultivars can vary across different growing locations, consequently impacting their nutrient profiles. This study employs untargeted metabolomics in conjunction with the quantification of fatty acids and amino acids to explore the influence of three different growing locations in Spain (namely Andalusia, Aragon and Asturias), on the nutritional characteristics of seeds of various pea cultivars. Significant interactions between cultivar and environment were observed, with 121 metabolites distinguishing pea profiles. Lipids, lipid-like molecules, phenylpropanoids, polyketides, carbohydrates, and amino acids were the most affected metabolites. Fatty acid profiles varied across locations, with higher C16:0, C18:0, and 18:1 n-9 concentration in Aragón, while C18:2 n-6 predominated in Asturias and C18:3 n-3 in Andalusia. Amino acid content was also location-dependent, with higher levels in Asturias. These findings underscore the impact of environmental factors on pea metabolite profiles and emphasize the importance of selecting pea cultivars based on specific locations and animal requirements. Enhanced collaboration between research and industry is crucial for optimizing pea cultivation for animal feed production.

Machine learning-enabled fatty acid quantification and classification of pork from autochthonous breeds using low-field 1H NMR spectroscopic data

Traditional pig breeds, known for their sustainability and superior meat quality, are experiencing growing consumer preference. The lipid fraction composition of these meats plays a fundamental role in their health benefits and excellent organoleptic properties. Accordingly, accurate characterisation of intramuscular fat is crucial for maintaining quality standards and combating fraudulent practices. This study employs benchtop nuclear magnetic resonance (NMR) spectroscopy to delineate the lipidic profiles of various cuts from two emblematic Spanish autochthonous pig breeds. The implementation of chemometric and machine learning models enabled the classification of pork samples based on cut and breed of origin. Moreover, this investigation pioneers the coupling of benchtop NMR with machine learning models for quantitative purposes, achieving precise quantification of polyunsaturated, monounsaturated and saturated fatty acids in intramuscular fat. This novel approach holds promise for enhancing the traceability and authentication of traditional pig products, fostering consumer confidence and promoting sustainable livestock practices.

Full nutritional spectrum and hazard risk assessment of high commodity value aquaculture species in Penang, Malaysia

Aquaculture is a rapidly growing food production sector that aims to alleviate global hunger, malnutrition, food security and nutrient deprivation. This study aimed to update the full spectrum of nutritional composition using aquaculture species. High commodity value aquaculture species in Penang, Malaysia have been shortlisted, followed by nutritional quantification and health risk estimation. Nutritional compositional analysis involved proximate analysis, and amino acid, fatty acid, vitamin, mineral and heavy metal quantification. Health risk assessment was conducted to determine whether the benefits were greater than the risks in the long term. Ten (10) aquaculture species, namely golden pomfret, golden snapper, seabass, bigeye trevally, red snapper, grouper, African catfish, blood cockles, white-leg shrimp and tiger prawn, have been studied extensively. The results showed that all species contained adequate protein and lipid contents, ranging from 13.6 % to 26.9 % and 0.4–15.8 %, respectively. The essential amino acid (EAA) to total amino acid (TAA) ratio of the culture species was comparable to the FAO reference values (50 %), with leucine and lysine demonstrating the most important EAAs. Fatty acid profiling revealed that both seabass and grouper were rich in polyunsaturated fatty acids, particularly docosahexaenoic acid. Golden pomfret and red snapper contained the greatest amounts of riboflavin and pyridoxine. The atherogenic and thrombogenic indices were within the permissible ratio (<1), except for those of African catfish. Notably, the arsenic (As) content in blood cockles exceeded the permissible criterion values. Hazard index (HI) assessment indicated that prolonged consumption of blood cockles, white-leg shrimp, tiger prawn, bigeye trevally and golden snapper demonstrated potent non-carcinogenic risk for both adults and children. This study provides an initial platform for informing consumers, stakeholders and policy makers about aquaculture nutritional benefits, consumption safety concern and health indicators.

Antimicrobial, antifungal, antioxidant activity and phytochemical investigation of fatty acids by GS/MS of raspberry (Rubus idaeus L.) shoot lipophilic extract

The aim of the study was to determine the content of fatty acids using GC-MS in the obtained extract, conduct a study of the antimicrobial, antifungal and antioxidant activities of Rubus idaeus shoot lipophilic extract. The quantification of fatty acids was accomplished through GC-MS, antioxidant activity was assessed by potentiometric method, antimicrobial and antifungal activity was determined by well method. The 18 compounds were identified by the GS-MS. The levulenic acid (64.47±0.08 mg/100 g), linoleic acid (8.50±0.08 mg/100 g) and linolenic acid (6.80±0.04 mg/100 g) dominated in the obtained lipophilic R. idaeus shoot extract. Bacillus subtilis (17.00±0.50 mm) was the most sensitive to lipophilic extract whereas P. vulgaris was the most resistant to the lipophilic extract. Moreover, Candida albicans was medium sensitive to lipophilic extract (13.50±0.50 mm). The antioxidant activity was 1.00 mmol equiv./mdry res, according to Maslov`s antioxidant level classification it has low level. The extract exhibited antimicrobial activity against all tested strains, with the most significant impact observed against B. subtilis. However, the obtained lipophilic extract showed a relatively low level of antioxidant activity. Consequently, the derivatives of fatty acids play a substantial role in the antimicrobial and antifungal effects, whereas their contribution to antioxidant activity appears to be limited.

High-throughput relative quantification of fatty acids by 12-plex isobaric labeling and microchip capillary electrophoresis - Mass spectrometry

BackgroundFatty acids (FAs) are essential cellular components and play important roles in various biological processes. Importantly, FAs produced by microorganisms from renewable sugars are considered sustainable substrates for biodiesels and oleochemicals. Their complex structures and diverse functional roles in biochemical processes necessitate the development of efficient and accurate methods for their quantitative analysis. ResultsHere, we developed a novel method for relative quantification of FAs by combining 12-plex isobaric N,N-dimethyl leucine-derivatized ethylenediamine (DiLeuEN) labeling and microchip capillary electrophoresis-mass spectrometry (CE-MS). This method enables simultaneous quantification of 12 samples in a single MS analysis. DiLeuEN labeling introduced tertiary amine center structure into FAs, which makes them compatible with the positive mode separation of commercial microchip CE systems and further improves the sensitivity. The CE separation parameters were optimized, and the quantification accuracy was assessed using FA standards. Microchip CE-MS detection exhibited high sensitivity with a femtomole level detection limit and a total analysis time within 8 min. Finally, the applicability of our method to complex biological samples was demonstrated by analyzing FAs produced by four industrially relevant yeast strains (Saccharomyces cerevisiae, Yarrowia lipolytica YB-432, Yarrowia lipolytica Po1f and Rhodotorula glutinis). The analysis time for each sample is less than 1 min. SignificanceThis work addresses the current challenges in the field by introducing a method that combines microchip-based capillary electrophoresis separation with multiplex isobaric labeling. Our method not only offers remarkable sensitivity and rapid analysis speed but also the capability to quantify fatty acids across multiple samples simultaneously, which holds significant potential for extensive application in FA quantitative studies in diverse research areas, promising an enhanced understanding of FA functions and mechanisms.

Behavioral comorbidities treatment by fecal microbiota transplantation in canine epilepsy: a pilot study of a novel therapeutic approach.

Anxiety and cognitive dysfunction are frequent, difficult to treat and burdensome comorbidities in human and canine epilepsy. Fecal microbiota transplantation (FMT) has been shown to modulate behavior in rodent models by altering the gastrointestinal microbiota (GIM). This study aims to investigate the beneficial effects of FMT on behavioral comorbidities in a canine translational model of epilepsy. Nine dogs with drug-resistant epilepsy (DRE) and behavioral comorbidities were recruited. The fecal donor had epilepsy with unremarkable behavior, which exhibited a complete response to phenobarbital, resulting in it being seizure-free long term. FMTs were performed three times, two weeks apart, and the dogs had follow-up visits at three and six months after FMTs. Comprehensive behavioral analysis, including formerly validated questionnaires and behavioral tests for attention deficit hyperactivity disorder (ADHD)- and fear- and anxiety-like behavior, as well as cognitive dysfunction, were conducted, followed by objective computational analysis. Blood samples were taken for the analysis of antiseizure drug (ASD) concentrations, hematology, and biochemistry. Urine neurotransmitter concentrations were measured. Fecal samples were subjected to analysis using shallow DNA shotgun sequencing, real-time polymerase chain reaction (qPCR)-based Dysbiosis Index (DI) assessment, and short-chain fatty acid (SCFA) quantification. Following FMT, the patients showed improvement in ADHD-like behavior, fear- and anxiety-like behavior, and quality of life. The excitatory neurotransmitters aspartate and glutamate were decreased, while the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) and GABA/glutamate ratio were increased compared to baseline. Only minor taxonomic changes were observed, with a decrease in Firmicutes and a Blautia_A species, while a Ruminococcus species increased. Functional gene analysis, SCFA concentration, blood parameters, and ASD concentrations remained unchanged. Behavioral comorbidities in canine IE could be alleviated by FMT. This study highlights FMT's potential as a novel approach to improving behavioral comorbidities and enhancing the quality of life in canine patients with epilepsy.

A new chemical derivatization reagent sulfonyl piperazinyl for the quantification of fatty acids using LC-MS/MS

In our previous study, a chemical derivatization reagent named 5-(dimethylamino) naphthalene-1-sulfonyl piperazine (Dns-PP) was developed to enhance the chromatographic retention and the mass spectrometric response of free fatty acids (FFAs) in reversed-phase liquid chromatography coupled with electrospray ionization-mass spectrometry (RPLC-ESI-MS). However, Dns-PP exhibited strong preferences for long-chain FFAs, with limited improvement for short- or medium-chain FFAs. In this study, a new series of labeling reagents targeting FFAs were designed, synthesized, and evaluated. Among these reagents, Tmt-PP (N2, N2, N4, N4-tetramethyl-6-(4-(piperazin-1-ylsulfonyl) phenyl)-1,3,5-triazine-2,4-diamine) exhibited the best MS response and was selected for further evaluations. We compared Tmt-PP with Dns-PP and four commonly used carboxyl labeling reagents from existing studies, demonstrating the advantages of Tmt-PP. Further comparisons between Tmt-PP and Dns-PP in measuring FFAs from biological samples revealed that Tmt-PP labeling enhanced the MS response for about 80 % (30/38) of the measured FFAs, particularly for short- and medium-chain FFAs. Moreover, Tmt-PP labeling significantly improved the chromatographic retention of short-chain FFAs. To ensure accurate quantification, we developed a stable isotope-labeled Tmt-PP (i.e., d12-Tmt-PP) to react with chemical standards and serve as one-to-one internal standards (IS). The method was validated for accuracy, precision, sensitivity, linearity, stability, extraction efficiency, as well as matrix effect. Overall, this study introduced a new chemical derivatization reagent Tmt-PP (d12-Tmt-PP), providing a sensitive and accurate option for quantifying FFAs in biological samples.

Feeding dogs a high-fat diet induces metabolic changes similar to natural aging, including dyslipidemia, hyperinsulinemia, and peripheral insulin resistance.

The goal of this study was to characterize changes induced by a high-fat diet in body composition, insulin levels and sensitivity, blood lipids, and other key biomarkers also associated with the metabolic dysfunction that occurs with natural aging. 24 male Beagle dogs, 3 to 7 years of age, of mixed castration status. Dogs were randomly assigned to continue twice daily feeding of the commercial adult maintenance diet (n = 12, including 2 intact) that they were previously fed or to a high-fat diet (12, including 2 intact) for 17 weeks between December 1, 2021, and April 28, 2022. Assessments included body composition (weight, body condition score, and adipose mass determined by deuterium enrichment), clinical chemistries, plasma fatty acid quantification, oral glucose tolerance test, and histology of subcutaneous and visceral adipose biopsy samples. The high-fat diet led to increased body weight, body condition score, fat mass and adipocyte size, hyperinsulinemia and peripheral insulin resistance, and elevations in serum lipids, including cholesterol, triglycerides, and several species of free fatty acids. Leptin levels increased in dogs fed a high-fat diet but not in control dogs. There were no significant changes in routine clinical chemistry values in either group. Feeding a high-fat diet for 17 weeks led to potentially deleterious changes in metabolism similar to those seen in natural aging in dogs, including hyperinsulinemia, insulin resistance, and dyslipidemia. A high-fat diet model may provide insights into the similar metabolic dysfunction that occurs during natural aging.

Optimized quantification of fatty acids in complex media using advanced analytical techniques.

Various medium formulations contain essential fatty acids at concentrations ranging from 10 to 100 mg/L. Accurate and precise lipid measurement in media is crucial for monitoring media quality and conducting studies on lipids in the context of cell culture. This study employed two-dimensional gas chromatography (GC × GC) analyses to offer enhanced resolution, sensitivity, and separation performance compared to GC. Quantification of fatty acid methyl esters (FAMEs) in a medium was conducted using GC × GC combined with a high-resolution mass spectrometer and flame ionization detector, considering potential interference from nonionic surfactant Tween 80, which was precipitated and removed by optimizing the concentration of cobalt thiocyanate (CTA) solution during pretreatment. This advanced analytical approach enabled identification of cis and trans isomers of identical molecular weights and determination of the location and number of double bonds in the same carbon number structure. Our analysis identified 36 FAMEs within the C6-C24 region, and a 5% CTA solution was optimal for efficient removal of Tween 80 during lipid extraction. Additionally, this advanced method minimized FAME contamination and loss during pretreatment, thereby significantly reducing the sample volume required to detect trace levels of FAMEs. This improvement led to a fatty acid recovery rate of 106% while maintaining the average relative standard deviation for the target FAMEs of about 3%. Our research paves the way for future investigation into medium quality control and the role of fatty acids in cell culture. This offers the possibility for economical and effective trace quantification of fatty acids in complex media.

TM7SF2-induced lipid reprogramming promotes cell proliferation and migration via CPT1A/Wnt/β-Catenin axis in cervical cancer cells

Cervical cancer poses a serious threat to women’s health globally. Our previous studies found that upregulation of TM7SF2, which works as an enzyme involved in the process of cholesterol biosynthesis expression, was highly correlated with cervical cancer. However, the mechanistic basis of TM7SF2 promoting cervical cancer progression via lipid metabolism remains poorly understood. Therefore, quantification of fatty acids and lipid droplets were performed in vitro and in vivo. The protein-protein interaction was verified by Co-IP technique. The mechanism and underlying signaling pathway of TM7SF2 via CPT1A associated lipid metabolism in cervical cancer development were explored using Western blotting, IHC, colony formation, transwell assay, and wound healing assay. This study reported that overexpression of TM7SF2 increased fatty acids content and lipid droplets both in vivo and in vitro experiments. While knockout of TM7SF2 obviously attenuated this process. Moreover, TM7SF2 directly bonded with CPT1A, a key enzyme in fatty acid oxidation, and regulated CPT1A protein expression in cervical cancer cells. Notably, the proliferation and metastasis of cervical cancer cells were elevated when their CPT1A expression was upregulated. Then, rescue assay identified that CPT1A overexpressed could enhance the cell viability and migration in TM7SF2-knockout cells. Furthermore, depletion of TM7SF2 significantly inhibited WNT and β-catenin proteins expression, which was enhanced by CPT1A-overexpressed. The proliferation and migration of cervical cancer cells were reversed in CPT1A-overexpressed cells with the treatment of MSAB, an inhibitor of Wnt/β-Catenin pathway. This study put forward an idea that TM7SF2-induced lipid reprogramming promotes proliferation and migration via CPT1A/Wnt/β-Catenin axis in cervical cancer, underlying the progression of cervical cancer.

Development and Validation of an LC-MS/MS Method for the Determination of Plasma and Red Blood Cell Omega Fatty Acids: A Useful Diagnostic Tool

Development and Validation of an LC-MS/MS Method for the Determination of Plasma and Red Blood Cell Omega Fatty Acids: A Useful Diagnostic Tool

A Two-Faced Gut Microbiome: Butyrogenic and Proinflammatory Bacteria Predominate in the Intestinal Milieu of People Living with HIV from Western Mexico.

HIV infection results in marked alterations in the gut microbiota (GM), such as the loss of microbial diversity and different taxonomic and metabolic profiles. Despite antiretroviral therapy (ART) partially ablating gastrointestinal alterations, the taxonomic profile after successful new ART has shown wide variations. Our objective was to determine the GM composition and functions in people living with HIV (PLWHIV) under ART in comparison to seronegative controls (SC). Fecal samples from 21 subjects (treated with integrase strand-transfer inhibitors, INSTIs) and 18 SC were included. We employed 16S rRNA amplicon sequencing, coupled with PICRUSt2 and fecal short-chain fatty acid (SCFA) quantification by gas chromatography. The INSTI group showed a decreased α-diversity (p < 0.001) compared to the SC group, at the expense of increased amounts of Pseudomonadota (Proteobacteria), Segatella copri, Lactobacillus, and Gram-negative bacteria. Concurrently, we observed an enrichment in Megasphaera and Butyricicoccus, both SCFA-producing bacteria, and significant elevations in fecal butyrate in this group (p < 0.001). Interestingly, gut dysbiosis in PLWHIV was characterized by a proinflammatory environment orchestrated by Pseudomonadota and elevated levels of butyrate associated with bacterial metabolic pathways, as well as the evident presence of butyrogenic bacteria. The role of this unique GM in PLWHIV should be evaluated, as well as the use of butyrate-based supplements and ART regimens that contain succinate, such as tenofovir disoproxil succinate. This mixed profile is described for the first time in PLWHIV from Mexico.