Docosahexaenoic Acid Research Articles

Protective Role of Eicosapentaenoic and Docosahexaenoic and Their N-Ethanolamide Derivatives in Olfactory Glial Cells Affected by Lipopolysaccharide-Induced Neuroinflammation

Neuroinflammation is a symptom of different neurodegenerative diseases, and growing interest is directed towards active drug development for the reduction of its negative effects. The anti-inflammatory activity of polyunsaturated fatty acids, eicosapentaenoic (EPA), docosahexaenoic (DHA), and their amide derivatives was largely investigated on some neural cells. Herein, we aimed to elucidate the protective role of both EPA and DHA and the corresponding N-ethanolamides EPA-EA and DHA-EA on neonatal mouse Olfactory Ensheathing Cells (OECs) after exposition to lipopolysaccharide (LPS)-induced neuroinflammation. To verify their anti-inflammatory effect and cell morphological features on OECs, the expression of IL-10 cytokine, and cytoskeletal proteins (vimentin and GFAP) was evaluated by immunocytochemical procedures. In addition, MTT assays, TUNEL, and mitochondrial health tests were carried out to assess their protective effects on OEC viability. Our results highlight a reduction in GFAP and vimentin expression in OECs exposed to LPS and treated with EPA or DHA or EPA-EA or DHA-EA in comparison with OECs exposed to LPS alone. We observed a protective role of EPA and DHA on cell morphology, while the amides EPA-EA and DHA-EA mainly exerted a superior anti-inflammatory effect compared to free acids.

Association between Dietary Antioxidants and Atherosclerotic Cardiovascular Disease in South Korea: Insights from a Comprehensive Cross-Sectional Analysis

Background/Objectives: The multifactorial nature of atherosclerotic cardiovascular disease (ASCVD) implicates genetic, environmental, and dietary habits. Antioxidants found in foods have garnered attention for their potential role in mitigating ASCVD risk by combating oxidative stress. This study seeks to confirm the findings of previous research through a large-scale cross-sectional analysis performed in a unique population with Korea National Health and Nutrition Examination Survey data to explore the association between the composite dietary antioxidant index (CDAI) and ASCVD prevalence among middle- and old-aged individuals in South Korea. Methods: This study includes data from 2016 to 2021. The CDAI was calculated based on nutrition intake, including zinc, beta-carotene, vitamin A, vitamin C, vitamin E, and docosahexaenoic acid. This cross-sectional analysis explored the relationship between the CDAI and ASCVD after adjusting for relevant covariates. Logistic regression models were employed, and subgroup analyses by sex were conducted to discern sex-specific effects. Results: A total of 19,818 individuals were analyzed, with 7.0% of them diagnosed with ASCVD. CDAI distribution and antioxidant analyses revealed higher CDAI levels in non-ASCVD individuals. Standardized antioxidant values increased across CDAI quartiles. Initially, a significant association (odds ratio [95% confidence interval]: 0.96 [0.94–0.99]) was found between the CDAI and ASCVD, which was attenuated after adjusting for covariates (1.0 [0.98–1.02]). Subgroup analyses by sex showed nuanced associations, with the CDAI potentially reducing the risk of ASCVD in men (0.71 [0.53–0.94]) while increasing it in women (1.4 [1.01–1.95]). Conclusions: This study provides valuable insights into the association between dietary antioxidant intake and the risk of ASCVD, highlighting sex-specific differences.

Investigating the effects of microplastics on the metabolism of Trematomus bernacchii from the ross sea (Antarctica)

Microplastic pollution is a growing environmental issue, even reaching remote areas like the Arctic and Antarctic, posing threats to biodiversity and food chains. The present research represents a pioneering endeavor aimed at exploring the relationship between lipids and microplastics in 20 wild specimens of Trematomus bernacchii from the Ross Sea (Antarctica). Fish were grouped in MPs-ingested and MPs-free based on whether they had ingested microplastics. Raman spectroscopy revealed that contaminated fish samples contained from one to three different types of polymeric fibers (1.4 items/specimen, ± = 0.7), specifically, polyester (PES), polypropylene (PP), and polyethylene terephthalate (PET). Gas chromatography-mass spectrometry (GC–MS) and gas chromatography-flame ionization detection (GC-FID) techniques were employed for the study of the lipid composition in term of fatty acids methyl esters (FAMEs). Fifty different FAME compounds were identified and quantified in the lipid fraction extracted from the muscle tissues of the selected fish samples. Polyunsaturated fatty acids (PUFAs) are the most abundant family of fatty acids in T. bernacchii species with eicosapentaenoic (C20:5ω3) and docosahexaenoic (C22:6ω3) acids as main components. In detail, PUFA class accounted for 46.78 ± 6.82% and 44.62 ± 4.86% of the total fatty acid composition in MPs-ingested and MPs-free groups, respectively. The contents of the monounsaturated fatty acids (MUFAs) and saturated fatty acids (SFAs) varied from 27.93% to 31.15% and from 24.23% to 25.05% in MPs-ingested and MPs-free fish samples, respectively. Based on Mann-Whitney test results (p < 0.05), there was no significant difference from a statistical point of view between two groups of fishes. Additionally, nutritional quality indices exhibited comparable values between groups. Results showed that no significant differences were found in the fatty acids distribution between the two groups. This indicates that the lipid composition of wild fish that are naturally exposed to plastic pollution remains unchanged and could still have beneficial effects on human health.

Maternal high-dose docosahexaenoic acid supplementation and neurodevelopment at 5 Years of preterm children

Background & AimsDocosahexaenoic acid (DHA) is the most abundant omega-3 fatty acid in the brain and is accumulated by the fetal brain during the last trimester of pregnancy. Our objective was to determine whether high-dose DHA supplementation during the neonatal period, vs. placebo, improves behavioral functioning at 5 years in children born very preterm. MethodsThis is a follow-up at 5 years corrected age of a subset of children who participated in a multicenter randomized controlled trial. The participants received a high-dose DHA supplementation, or a placebo, through maternal breast milk until 36 weeks’ postmenstrual age. Primary outcome was child behavioral functioning, assessed by the Total Difficulties Score from the Strengths and Difficulties Questionnaire (SDQ). Secondary outcomes included behavioral scores from the SDQ, executive functions assessment and global developmental performance. Neurodevelopmental outcomes were assessed through interviews with parents. Mean differences between DHA and placebo groups were estimated using mixed linear models. Subgroup analyses were conducted for sex and gestational age (GA) at birth. ResultsAmong 177 eligible children, 132 (74.6%) completed neurodevelopmental assessment at 5 years (DHA, N=64, placebo, N=68). Total Difficulties Score did not differ between the DHA and placebo groups (mean differences, -0.9 [95% confidence interval, -2.7 to 0.8], P=0.30), nor any of the secondary outcomes. There was no significant interaction between treatment groups and sex, nor GA, for the primary outcome. However, significant interactions between treatment groups and sex or GA were found for some secondary outcomes. ConclusionsIn very preterm infants, high-dose DHA supplementation did not improve behavioral functioning at 5 years. Clinical Trial RegistrationClinicalTrials.gov, NCT02371460, https://clinicaltrials.gov/study/NCT02371460

Enhancing fatty acid and omega-3 production in Schizochytrium sp. using developed safe-harboring expression system

BackgroundSchizochytrium, a group of eukaryotic marine protists, is an oleaginous strain, making it a highly promising candidate for the production of lipid-derived products such as biofuels and omega-3 fatty acids. However, the insufficient advancement of genetic engineering tools has hindered further advancements. Therefore, the development and application of genetic engineering tools for lipid enhancement are crucial for industrial production.ResultsTransgene expression in Schizochytrium often encounters challenges such as instability due to positional effects. To overcome this, we developed a safe-harbor transgene expression system. Initially, the sfGFP gene was integrated randomly, and high-expressing transformants were identified using fluorescence-activated cell sorting. Notably, HRsite 2, located approximately 3.2 kb upstream of cytochrome c, demonstrated enhanced sfGFP expression and homologous recombination efficiency. We then introduced the 3-ketoacyl-ACP reductase (KR) gene at HRsite 2, resulting in improved lipid and docosahexaenoic acid (DHA) production. Transformants with KR at HRsite 2 exhibited stable growth, increased glucose utilization, and a higher lipid content compared to those with randomly integrated transgenes. Notably, these transformants showed a 25% increase in DHA content compared to the wild-type strain.ConclusionThis study successfully established a robust homologous recombination system in Schizochytrium sp. by identifying a reliable safe harbor site for gene integration. The targeted expression of the KR gene at this site not only enhanced DHA production but also maintained growth and glucose consumption rates, validating the efficacy of the safe-harbor approach. This advancement in synthetic biology and metabolic engineering paves the way for more efficient biotechnological applications in Schizochytrium sp.

Insights into the methodological perspectives for screening polyunsaturated fatty acids-containing bacteria.

Polyunsaturated fatty acids (PUFA) are vital molecules in the pharmaceutical, medical, and nutritional industries. Exploration of bacterial strains capable of producing significant amounts of PUFAs offers a promising avenue for biotechnological applications and industrial-scale production. However, an extensive screening of several samples from diverse sources is highly needed to identify a potential strain. The present study provides the results of the evaluation of 15 different screening methodologies (including changes in existing protocols in terms of reagent concentration, incubation temperature and time) for identifying PUFA-producing bacteria in comparison to the gold standard method (Gas chromatography-mass spectrometry), for the first time. The results determined the most effective techniques for each critical PUFA, leading to an optimized screening process that saves time and resources. The H2O2 plate assay using 0.5% or 1% H2O2 for 72 & 96h of incubation at 15 °C consistently outperformed others for finding bacteria containing total nutritionally important long chain-PUFA (LC-PUFA), linoleic acid, and arachidonic acid. Whereas the 2,3,5-triphenyl tetrazolium chloride broth assay at 10-15 °C was the most effective and semiquantitative screening methodology for eicosapentaenoic acid (EPA) and alpha-linolenic acid-containing bacteria. Apart from the methodological perspectives, the study also revealed certain potential strains to be targeted in the ongoing research on PUFA-containing bacteria. Further, the manuscript forms the first report on the presence of docosahexaenoic acid (DHA) in Shewanella decolorationis, EPA in Psychrobacter maritimus and Micrococcus aloeverae, and both EPA and DHA in Arthrobacter rhombi. Altogether, the paper generates several thought-provoking insights on the methodological perspectives and identifies potential PUFA-containing bacteria with practical applications in future bacteria-based PUFA research.

Macronutrient Intake during Complementary Feeding in Very Low Birth Weight Infants Comparing Early and Late Introduction of Solid Foods: A Secondary Outcome Analysis.

Background/Objectives: Very low birth weight (VLBW) infants may require enhanced nutrition, even during complementary feeding. However, there are limited data on macronutrient intake during this period, particularly concerning the individual timing of the introduction of solid foods in a representative VLBW infant population. Methods: This prospective observational study analyzed macronutrient intake in VLBW infants with a gestational age < 32 weeks based on whether solid foods were introduced early (<17 weeks corrected age (CA)) or late (≥17 weeks corrected age) Nutritional intake was analyzed using a 24 h recall at 6 weeks CA and 3-day dietary records at 12 weeks, 6, 9, and 12 months CA. Results: In total, 115 infants were assigned to the early and 82 to the late group. The timing of solid food introduction did not affect macronutrient intake, except for a lower fat and higher carbohydrate intake (% of energy) in the early group at 12 weeks and 6 months CA: early vs. late, fat-12 weeks: 47.0% vs. 49.0%, 6 months: 39.2% vs. 43.3%; carbohydrates-12 weeks: 44.9% vs. 43.2%, 6 months: 51.3% vs. 48.0%. Apart from docosahexaenoic acid (DHA) and arachidonic acid (AA), dietary intake recommendations were met in both groups. While nutrient intakes varied significantly between breastfed and formula-fed infants, those with comorbidities exhibited similar nutrient intake levels compared to those without. Conclusions: Our findings suggest adequate macronutrient intakes in VLBW infants irrespective of the timing of solid introduction. However, there is a notable need to enhance dietary intakes of DHA and AA. Future research is crucial to assess whether current nutrient intakes are sufficient for VLBW infants with comorbidities.

Eicosapentaenoic Acid and Medium-Chain Triacylglycerol Structured Lipid Supplementation Improves Muscular Endurance Exercise Performance and Reduces Muscle Fatigue in Young Healthy Male

Objective Structured lipids containing medium-chain fatty acids, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) from fish oil (EPA/MCT) and it improve overall endurance. However, their effects on local muscle endurance remain unclear. This study aimed to investigate the effects of EPA/MCT intake on muscle endurance during leg extension exercises and subsequent muscle fatigue. Methods This 8-week, placebo-controlled, randomized, double-blind parallel-group trial involved 20 healthy young men. Subjects were randomly assigned to an EPA/MCT group (10 subjects) and a placebo group (PL group, 10 subjects). They consumed 4560 mg of the test food daily for 8 wk before an exercise intervention. The EPA/MCT supplement contained 600 mg of EPA, 260 mg of DHA, and 1890 mg of MCT (1110 mg of octanoic acid and 780 mg of decanoic acid). After the 8-week intervention, the subjects performed four sets of leg extension exercises at 40% of their body weight until exhaustion. The number of repetitions per set and the total repetitions across the four sets served as indicators of muscle endurance. Isometric maximum voluntary contraction (MVC), range of motion, circumference, muscle thickness, echo intensity, and muscle stiffness were measured before and after exercise to assess muscle fatigue. Results There was no difference between the groups in the number of repetitions for sets 1–4 or in the total number of repetitions during the leg extension exercise. In the EPA/MCT group, a significant negative correlation was observed between the increase in the blood EPA/AA ratio and the rate of decrease in the number of repetitions. Additionally, the decrease in MVC after exercise was significantly smaller in the EPA/MCT group than in the PL group. Conclusion Our study results suggest that an 8-week intake of EPA/MCT can increase the blood EPA/AA ratio and improve muscle endurance performance. Additionally, EPA/MCT intake reduces muscle fatigue after exercise.

Evolution of the lipoxygenase gene family in Pyropia haitanensis and its response to biotic and abiotic stresses

Macroalgae contain a chemically diverse and unique set of oxylipins that derived from lipid peroxidation play pivotal roles in response to various environmental stresses in the intertidal zone. Specific enzymes involved in biosynthesizing these oxylipins remain largely unknown. Lipoxygenase (LOX) is a key enzyme in the oxylipin biosynthetic pathway. Here, six homologous LOX genes were identified in Pyropia haitanensis. To investigate the evolutionary status of the LOX gene family in red algae and its role in the adaptation of P. haitanensis to the intertidal environment, this study analyzed the evolution, structure, and enzymatic activities of these genes. Phylogenetic analysis and homologous sequence alignment indicated that the P. haitanensis LOX genes had two evolutionary origins. PhLOX and PhLOX2 possess an SRPBCC domain at the N-terminus that may have been acquired from marine bacteria through horizontal gene transfer, while PhLOX3–6 may have been acquired from cyanobacteria. Cis-Regulatory analysis revealed numerous elements upstream of the PhLOX genes associated with abiotic stress and hormone regulation, as well as binding sites for defense-related transcription factors. The in-vitro enzymatic activity of PhLOXs was detected, revealing that PhLOX and PhLOX2 exhibited the highest activities. PhLOX3–6 showed a substrate preference for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), but it had lower activity. The transcriptional expression levels of the six PhLOX genes varied under different biotic and abiotic stresses. For example, all PhLOX genes were significantly upregulated after mechanical damage, while PhLOX5 responded to low temperatures. In summary, P. haitanensis acquired a series of LOX genes from marine bacteria and cyanobacteria during its evolutionary history, among which PhLOX and PhLOX2 were selected as the likely primary functional genes. Its functional diversity to cope with various environmental stresses is achieved through the complex transcriptional regulation of PhLOX genes.

Capillary Blood Docosahexaenoic Acid Levels Predict Electrocardiographic Markers in a Sample Population of Premenopausal Women.

Introduction: The relationship between blood N-3 polyunsaturated fatty acid (PUFA) levels and cardiovascular health is known, but direct evidence that N-3 PUFA levels influence electrocardiographic (ECG) parameters is non-existent. In the study described herein, we investigated the relationship between anthropometric biomarkers and capillary blood PUFAs with ECG outputs in a sample population of healthy pre-menopausal women. Method: Twenty-three consenting females were recruited, with the study power analysis sufficiently demonstrated. Food intake, anthropometric and cardiovascular parameters were obtained. Capillary blood was collected for fatty acid chromatographic analysis. Results: Body mass index, haematocrit, heart rate (HR), mean arterial pressure (MAP) and ECG readings all fell within healthy ranges. Principal component analysis-mediated correlations were carried out controlling for combined Components 1 (age, body fat % and waist-to-hip ratio) and 2 (height, HR and MAP) as control variables. Docosahexaenoic acid (DHA) unequivocally decreased the QRS area under the curve (AUC-QRS) regardless of the impact of control variables, with each unit increase in DHA corresponding to a 2.3-unit decrease in AUC-QRS. Mediation analysis revealed a significant overall effect of DHA on AUC-QRS, with the impact of DHA on R wave amplitude accounting for 77% of the total observed effect. Discussion: Our new findings revealed an inverse relationship between AUC-QRS with capillary blood DHA, suggesting that the association between ventricular mass and its QRS depolarising voltage is mediated by DHA. Our findings bridge a knowledge gap on the relationship between ventricular mass and ventricular efficiency. Further research will confirm whether the relationship identified in our study also exists in diseased patients.

Docosahexaenoic acid (DHA) is a driving force regulating gene expression in bluefin tuna (Thunnus thynnus) larvae development

This study elucidated the role of DHA-modulated genes in the development and growth of Atlantic bluefin tuna (Thunnus thynnus) larvae ingesting increasing levels of DHA in their rotifer prey. The effect of feeding low, medium, and high rotifer (Brachionus rotundiformis) DHA levels (2.0, 3.6 and 10.9 mg DHA g−1 DW, respectively) was tested on 2–15 days post hatching (dph) bluefin tuna larvae. Larval DHA content markedly (P < 0.05) increased in a DHA dose-dependent manner (1.5, 3.9, 6.1 mg DHA g−1 DW larva, respectively), that was positively correlated with larval prey consumption and growth (P < 0.05). Gene ontology enrichment analyses of differentially expressed genes (DEGs) demonstrated dietary DHA significantly (P < 0.05) affected different genes and biological processes at different developmental ages. The number of DHA up-regulated DEGs was highest in 10 dph larvae (491), compared to 5 (12) and 15 dph fish (34), and were mainly involved in neural and synaptic development in the brain and spinal cord. In contrast, DHA in older 15 dph larvae elicited fewer DEGs but played critical roles over a wider range of developing organs. The emerging picture underscores the importance of DHA-modulated gene expression as a driving force in bluefin tuna larval development and growth.

8077 Use of Lovaza for Management of Hypertriglyceridemia in Pregnancy

Abstract Disclosure: J. Segarra-Villafane: None. L.R. Sepulveda-Garcia: None. L. El Musa Penna: None. I.C. Arroyo: None. Z. Maisonet -Feliciano: None. W. Medina-Torres: None. M. Alvarado: None. M. Ramirez: None. L.A. Gonzalez-Rodriguez: None. Severe hypertriglyceridemia in pregnancy increases the risk of health complications to both mother and fetus, including preeclampsia, hyperviscosity syndrome and life-threatening pancreatitis. Several case reports include management of hypertriglyceridemia in pregnancy with medical nutrition therapy, omega fatty acids and gemfibrozil but is controversial. There are no formal clinical guidelines for management of severe hypertriglyceridemia in pregnancy which presents a challenge for physicians. Lovaza is a combination of ethyl esters of omega 3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) indicated as an adjunct to diet to reduce triglycerides (TG) levels in adult patients with severe hypertriglyceridemia. Lovaza is Category C (risk cannot be ruled out) and should be used in pregnancy only if the potential benefit justifies the potential risk to the fetus. Case of a 40 years old female G3P1A1 with IUP 25 WGA with history of hypothyroidism, dyslipidemia and episode of acute pancreatitis in 2016 due to hypertriglyceridemia, who was referred to clinics due to uncontrolled lipids and thyroid disease. Laboratories included fasting serum triglycerides (TG) 4357mg/dL (40-150mg/dL), total cholesterol 525mg/dL (120-200), HDL 56mg/dL, LDL invalid due to elevated TG. Thyroid function test revealed TSH of 161uIU/mL (0.43-2.91) and FT4 of 0.24ng/dL (0.8-1.8). HbA1c, lipase and amylase within range. Vital signs were normal. Patient was asymptomatic, no abdominal complaints or hypothyroidism-related symptoms, but hospital admission was recommended due to high risk of pancreatitis and uncontrolled hypothyroidism. She was started on Synthroid 220 mcg IV daily for hypothyroidism and was placed NPO until nutritionist evaluation. Triglycerides levels improved with conservative interventions and low-fat diet. However, TG persisted above 2000mg/dL and, after discussing risks and benefits with patient, pharmacological therapy with Lovaza was started to reach goal of TG less than 1000 mg/dL and reduce potential risk of pancreatitis. Patient agreed to start Lovaza 2gm AM and 2gm PM. After initiation of Lovaza, in addition to dietary modifications and hypothyroidism management, laboratories showed significant improvement in TG levels from 3569 down to 1778 mg/dL. Patient was discharged on Lovaza 2gm oral BID and Synthroid 200mcg oral daily. One month later, TG had decreased to 557mg/dL (0-150) and thyroid tests showed TSH at 0.305mIU/mL and FT4 at 1.71ng/dL. Patient tolerated medications well without side effects and had an uncomplicated pregnancy and delivery. This case demonstrates that severe hypertriglyceridemia during pregnancy can be managed effectively and, more importantly, in a safe manner with Lovaza, in addition to dietary fat restriction. However, additional research is needed if its use is to be included in future pregnancy management guidelines. Presentation: 6/2/2024

Enhancing DHA supplementation adherence: A Bayesian approach with finite mixture models and irregular interim schedules in adaptive trial designs.

Docosahexaenoic acid (DHA) supplementation has proven beneficial in reducing preterm births. However, the challenge lies in addressing nonadherence to prescribed supplementation regimens-a hurdle that significantly impacts clinical trial outcomes. Conventional methods of adherence estimation, such as pill counts and questionnaires, usually fall short when estimating adherence within a specific dosage group. Thus, we propose a Bayesian finite mixture model to estimate adherence among women with low baseline red blood cell phospholipid DHA levels (<6%) receiving higher DHA doses. In our model, adherence is defined as the proportion of participants classified into one of the two distinct components in a normal mixture distribution. Subsequently, based on the estimands from the adherence model, we introduce a novel Bayesian adaptive trial design. Unlike conventional adaptive trials that employ regularly spaced interim schedules, the novelty of our proposed trial design lies in its adaptability to adherence percentages across the treatment arm through irregular interims. The irregular interims in the proposed trial are based on the effect size estimation informed by the finite mixture model. In summary, this study presents innovative methods for leveraging the capabilities of Bayesian finite mixture models in adherence analysis and the design of adaptive clinical trials.

Association of circulating fatty acids with cardiovascular disease risk: Analysis of individual-level data in three large prospective cohorts and updated meta-analysis.

Associations of saturated and unsaturated fatty acids (FAs) with cardiovascular disease (CVD) remain controversial. We therefore aimed to investigate the prospective associations of objectively measured FAs with CVD, including incident coronary heart disease (CHD) and stroke, as well as CVD mortality. Circulating FA concentrations expressed as the percentage of total FAs were assayed in 172,891 participants without prior vascular disease at baseline from the European Prospective Investigation into Cancer and Nutrition-CVD (EPIC-CVD) (7,343 CHD; 6,499 stroke), UK Biobank (1,825; 1,474), and INTERVAL (285; 209) cohort studies. Hazard ratio (HR) per 1-standard deviation (SD) higher FA concentrations was estimated using Cox regression models and pooled by random-effects meta-analysis. Systematic reviews with meta-analysis published by 6 May 2023 on associations between FAs and CVDs were systematically searched and updated meta-analyses using random-effects model were conducted. Evidence from randomized controlled trials (RCTs) was also summarized. Higher concentrations of total saturated FAs (SFAs) were associated with higher cardiovascular risks in the combined analysis, with differential findings noted for SFA subtypes in further analysis restricted to EPIC-CVD: positive associations for even-chain SFA [HR for CHD 1.24 (95% CI: 1.18-1.32); stroke 1.23 (1.10-1.38)] and negative associations for odd-chain [0.82 (0.76-0.87); 0.73 (0.67-0.78)] and longer-chain [0.95 (0.80-1.12); 0.84 (0.72-0.99)] SFA. In the combined analysis, total n-3 polyunsaturated FA (PUFA) [0.91 (0.85-0.97)], including docosahexaenoic acid (DHA) [0.91 (0.84-0.98)], was negatively associated with incident CHD risk. Similarly, total n-6 PUFA [0.94 (0.91-0.98)], including linoleic acid (LA) [0.89 (0.83-0.95)], was negatively associated with incident stroke risk. By contrast, more detailed analyses in EPIC-CVD revealed that several downstream n-6 PUFAs of LA were positively associated with CHD risk. Updated meta-analyses of 37 FAs including 49 non-overlapping studies, involving between 7,787 to 22,802 CHD and 6,499 to 14,221 stroke cases, showed broadly similar results as our combined empirical analysis and further suggested significant inverse associations of individual long-chain n-3 PUFAs and LA on both CHD and stroke. The findings of long-chain n-3 PUFAs were consistent with those from published RCTs on CHD despite insufficient evidence in monotherapy, while RCT evidence remained unclear for the rest of the explored FAs. Our study provides an overview of the most recent evidence on the associations between objectively measured FAs and CVD outcomes. Collectively, the data reveals notable differences in associations by SFA subtypes and calls for further studies, especially RCTs, to explore these links.

Pharmacodynamic insights into maresin 1: Enhancing flap viability via the keap1/Nrf2 axis to control ROS-driven apoptosis and ferroptosis

Random flaps are widely used in tissue reconstruction, but the high incidence of flap necrosis after operation remains a significant challenge. Maresin 1 (MaR1), a mediator derived from docosahexaenoic acid, has been shown to have significant effects in resolving inflammation and promoting tissue regeneration. This study investigated the role of MaR1 in the survival of random flaps. Histological analysis, laser Doppler blood flow imaging, Masson trichrome staining, and survival area analysis were used to assess the viability of the flaps. Apoptosis, ferroptosis, oxidative stress, angiogenesis, and the underlying mechanisms were explored by examining the expression of specific molecules using immunofluorescence, western blotting, and other immunological and molecular biology techniques. The findings demonstrated that MaR1 could improve flap lifespan by significantly reducing oxidative stress, apoptosis, and ferroptosis, as well as by enhancing angiogenesis. The Keap1-Nrf2 pathway was upregulated by MaR1, which inhibited ROS-mediated apoptosis and ferroptosis. The protective effect of MaR1 on flap survival was abolished by ML385. Our findings indicate that MaR1 could be a novel therapeutic agent for enhancing flap treatment outcomes.

Phospholipids, Sphingolipids, and Cholesterol-Derived Lipid Mediators and Their Role in Neurological Disorders.

Neural membranes are composed of phospholipids, sphingolipids, cholesterol, and proteins. In response to cell stimulation or injury, the metabolism of lipids generates various lipid mediators, which perform many cellular functions. Thus, phospholipids release arachidonic acid or docosahexaenoic acid from the sn-2 position of the glycerol moiety by the action of phospholipases A2. Arachidonic acid is a precursor for prostaglandins, leukotrienes, thromboxane, and lipoxins. Among these mediators, prostaglandins, leukotrienes, and thromboxane produce neuroinflammation. In contrast, lipoxins produce anti-inflammatory and pro-resolving effects. Prostaglandins, leukotrienes, and thromboxane are also involved in cell proliferation, differentiation, blood clotting, and blood vessel permeability. In contrast, DHA-derived lipid mediators are called specialized pro-resolving lipid metabolites (SPMs). They include resolvins, protectins, and maresins. These mediators regulate immune function by producing anti-inflammatory, pro-resolving, and cell protective effects. Sphingolipid-derived metabolites are ceramide, ceramide1-phosphate, sphingosine, and sphingosine 1 phosphate. They regulate many cellular processes, including enzyme activities, cell migration and adhesion, inflammation, and immunity. Cholesterol is metabolized into hydroxycholesterols and 7-ketocholesterol, which not only disrupts membrane fluidity, but also promotes inflammation, oxidative stress, and apoptosis. These processes lead to cellular damage.

A-066 Does essential fatty acid deficiency alter ethanolamine plasmalogen concentrations in red blood cells? Implications for clinical testing

Abstract Background Plasmalogens are glycerophospholipids characterized by a fatty alcohol at the sn-1 position, a polyunsaturated fatty acid at the sn-2 position, most commonly docosahexaenoic acid (DHA) or arachidonic acid, and a polar head group at the sn-3 position, mainly phosphoethanolamine. Markedly reduced plasmalogens are seen in peroxisome biogenesis defects, including Zellweger spectrum disorders (ZSD), since the first two steps of plasmalogen synthesis occur in peroxisomes. Disruption of the peroxisomal β-oxidation pathway also causes DHA deficiency in ZSD patients. Since plasmalogens are composed of fatty acids, the deficiency of DHA or other essential fatty acids could further decrease the plasmalogen level in ZSD patients and lower the results of plasmalogen testing in patients without ZSD. To test this hypothesis, we evaluated the impact of essential fatty acid (EFA) deficiency on plasmalogens in patients with ZSD and individuals with EFA deficiency due to unrelated causes. Methods Red blood cell (RBC) samples were obtained from individuals without ZSD referred to ARUP laboratories for clinical EFA testing (n = 80; age range: birth - 89 years), and from patients with ZSD (n = 7; age range birth - 9 years). Three ZSD patients were older and with a less severe phenotype. A total of 18 intact ethanolamine plasmalogen species were extracted from packed RBCs and quantified using a XEVO TQ-XS Mass Spectrometer in conjunction with Ultra-High Performance Liquid Chromatography (Waters). Total plasmalogen values were also calculated; plasmalogen deficiency was defined as ≤60% of age-matched control median. Fatty acids (FAs) were quantified by gas chromatography negative chemical ionization-mass spectrometry (GC-NCI-MS). EFA deficiency was diagnosed by increased Triene/Tetraene ratio (Mead acid/Arachidonic acid). Data was analyzed using Prism v.8.3.0 software (La Jolla, CA). The study was approved by the Institutional Review Board of the University of Utah. Results Out of the 80 samples from individuals without ZSD referred for EFA testing, 18% had normal fatty acid profile, 57% were receiving dietary supplements and 25% had elevated Triene/Tetraene ratio suggesting EFA deficiency. EFA deficiency was accompanied by low DHA only in two individuals without ZSD, whereas it was present in most of the samples from ZSD patients evaluated (5/7). None of the individuals without ZSD were deficient in plasmalogens. Moreover, when compared to individuals without ZSD with a normal FA profile, there was no significant difference observed with EFA deficiency or FA supplementation. Although our ZSD cohort is small, no correlation between EFA deficiency and plasmalogen levels was also seen in ZSD: 2 patients with milder phenotype had EFA deficiency but normal plasmalogens, 2 patients had EFA deficiency and low plasmalogens, 2 patients with low plasmalogens had low DHA without EFA deficiency, and 1 patient had low plasmalogens but normal EFA. Conclusions Plasmalogen quantitation in RBCs by LC-MS/MS is not significantly affected by EFA deficiency. Although more data is warranted to elucidate the correlation between plasmalogen levels and ZSD phenotype, plasmalogens can be within normal reference intervals in ZSD regardless of EFA status.

B-350 The application of MALDI-TOF imaging to biomarker discovery in a mouse model of papillomavirus associated tumorigenesis

Abstract Background Human papillomavirus (HPV) can cause infection and dysplasia in cutaneous epithelium of immunodeficient patients. An animal model has been developed to study HPV-associated tumorigenesis using Nude mouse (Foxn1nu) and mouse papillomavirus (MmuPV1). This study applies MALDI-TOF imaging to compare the expression pattern of small molecules between viral-infected dysplastic mouse skin and mock-infected control tissue. Three sets of molecules showing distinct patterns are identified in correlation with the upregulation of an enzyme in related biosynthetic pathway. Methods Nude mice were wounded on tail and inoculated with MmPV1 suspension (1×109 viral DNA genomes per infected site) or PBS 24 hours later for treatment and control group, respectively. Tail skins were harvested 4 weeks post-infection and processed for cryosectioning. Tissue sections were analyzed using timsTOF flex (Bruker) under negative ion mode with 9-aminoacridine as the matrix. Viral infection (MmuPV1-L1 antigen) and enzyme expression (3'-phosphoadenosine 5'-phosphosulfate synthetase 1, PAPSS1) were assessed by immunofluorescence on adjacent sections. Results Three peaks demonstrated distinct distribution patterns in mouse skin. One peak at m/Z = 465 was identified as cholesterol sulfate (CS) by tandem MS and comparing with the standard. CS was enriched in keratinocytes, hair follicles, and blood vessels in normal skins, and it increased in dysplastic skins with enhanced angiogenesis. The combined peaks at m/Z = 327 &amp; 329 were tentatively identified as docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA). These long chain polyunsaturated fatty acids (LC-PUFAs) were present in the dermis as foci partially co-localized with CS, and they also increased in virus-infected skins. The peak at m/Z = 426 may represent a combination of ADP and APS. Although these two isobars cannot be separated by tandem MS, APS was revealed by the presence of sulfur isotopes at expected natural abundance (34S and 32S). They were present at a low level in the epidermis of normal skins but significantly increased after virus infection. APS and ADP were closely associated with the proliferating cells, and the signal intensity also increased with the degree of tissue dysplasia. As a single factor, APS &amp; ADP demonstrated a fair differential power for MmuPV1-induced dysplastic skin and normal control (AUC of ROC curve = 0.75). The bifunctional enzyme PAPSS1 catalyzes the conversion of ATP to 3’-phosphonato-5’-adenyl sulfate (PAPS) via APS as an intermediate. Interestingly, immunofluorescence showed the expression of PAPSS1 also increased significantly with the severity of tissue dysplasia and MmuPV1 infection. Conclusions MALDI-TOF imaging reveals three sets of molecules with distinct patterns in mouse skin, and their levels are increased during MmuPV1-induced epithelium dysplasia. As a precursor of the common sulfate donor (PAPS), increased APS and ADP in the epidermis of virus-infected skins indicate an upregulation of sulfation pathway and adenine metabolism. This finding is supported by enhanced expression of enzyme PAPSS1 and an increased sulfation reaction product, CS. In addition, DHA and DPA are also increased in the dermis of virus-infected skins, implying a disturbance in LC-PUFA metabolism. This study demonstrates combined use of MALDI-TOF imaging and immunostaining in the search of novel biomarkers of HPV-associated tumorigenesis.

Differences in quality between canned and pouched tuna packed with various media

Canned and pouched tuna products may be present in different liquid media such as brine, different types of oil and sauces in the markets. Limited reported information about the pouched tuna products and the differences between them in different liquid media available in Turkish markets. pH values of CW and PW were lower than the others. TBA values of all groups were found to be below the limits for development of an objectionable odor/taste. Omega 6/omega3 ratio was determined very high in CS and PS. On the other hand, higher content of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were found in CW and PW when compared with others (CO, CS, PO, PS). Heavy metal contents of tuna products, they were found to be safe according to limits for arsenic, mercury, lead and cadmium levels. L* values of all canned tunas (CO, CS, CW) were significantly higher than the pouched tunas (PO, PS, PW). b* values of both CW and PW were lower than the other groups. CW got the significantly lowest overall quality, color and taste scores when compared with the others. As a matter of fact, pouched tuna products especially PW were recommended for health purposes.

Biochemical analysis and fertilization success of Cirrhinus mrigala ova during induced spawning

Fish egg quality is very crucial in aquaculture sector for production of healthy seed. Egg yolk is an energy reservoir for growth and development of embryo. This study evaluated the biochemical composition and quality of Cirrhinus mrigala eggs at three different hatchery sites of Pakistan, (Site 1= Fish Seed Hatchery, District Pakpattan; Site 2= Sidhuwan Hatchery, Head Balloki, District Kasur; Site 3= Chenab Fish Hatchery, Rangpur, District Muzaffargarh) during induced breeding. For this, a total of 36 (18 males and 18 females) fish brooders, 12 (06 males and 06 females) from each site were utilized. Fatty acids analysis revealed significant differences among three different sites. Saturated fatty acids i.e., palmitic acid (C16:0) and stearic acids (C18:0) were higher at site 2 compared to the others sites. Monounsaturated fatty acids i.e., Oleic acid (C18:1) and polyunsaturated fatty acids i.e., Docosahexaenoic acid (DHA) (C22:6) exhibited considerably greater values at site 2 than those of other two sites. Egg mineral contents unveiled remarkable differences, particularly at site 2 indicating significantly higher mineral contents except copper (Cu) in comparison to the other sites. Significant variation exists in fertilization and hatching rates during induced spawning, with the highest values recorded at site 2. It is concluded that biochemical composition of egg especially fatty acid profile and mineral content greatly influences the embryonic development and hatching success of farm reared Cirrhinus mrigala.