Triglyceride Concentrations Research Articles

The Assessment of Anthropometric Measures and Changes in Selected Biochemical Parameters in Obese Children in Relation to Blood Lead Level.

Our paper draws attention to the impact of lead (Pb) on the specificity of obesity development in children exposed to environmental pollution. An advantage of this paper is the homogeneous study group comprising children of identical age from a single geographic region. Moreover, while the influence of environmental toxins on adults has been extensively explored, this study delves into pediatric populations, which have yet to receive comprehensive scrutiny within the scientific literature. Initially, a group of 136 obese children (the research program lasted three consecutive years: 2016, 2017, and 2018) living in the north-western region of Poland, from whom biochemical tests and auxological data were obtained, were enrolled for analysis. Blood lead levels (BLLs) were determined in 115 children. The age of the children ranged from 7.1 to 10.4 years. The body mass index (BMI) of children averaged 21.5 ± 2.2. The results showed that a large proportion of the participants had BLLs above the threshold for Pb. BLLs ≤ 5 µg/dL (considered safe for children and pregnant women) were found in over 70% of the participants, with BLLs in the range of 5.01-10.00 µg/dL in over 26% of the children, and concentrations > 10 µg/dL (considered toxic threshold for adults) in nearly 2% of the children. The results of our research revealed a positive association between BLLs and average systolic and diastolic blood pressure in the studied children. Moreover, we found a negative correlation between BLLs and absolute fat tissue content and triglyceride concentration. Among the included biochemical factors, only insulin demonstrated a statistically significant relationship with fat mass. This result suggests that early carbohydrate metabolism disorders in overweight children involve decreased peripheral tissue insulin sensitivity. Lead exposure may significantly contribute to the development of hypertension, insulin resistance, and glucose metabolism disorders in overweight and obese children. It is essential to implement multidirectional actions to increase awareness of the harmful effects of xenobiotic exposure, including lead, in order to prevent early-life exposure.

Clinical application of centrifugal-membrane hybrid plasmapheresis in the treatment of hyperlipidemia.

This study aimed to clarify the clinical application of centrifugal-membrane hybrid plasmapheresis (CMHP) in the treatment of hyperlipidemia. A retrospective study was conducted on 48 patients who were diagnosed with hyperlipidemia and had received CMHP treatment. Serum total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were monitored, and adverse reactions to the treatment were observed. Forty-eight patients with hyperlipidemia received CMHP over 59 sessions. The average age of the 48 patients with hyperlipidemia, including 32 males (66.67%) and 16 females (33.33%), was 44.23 ± 12.02 years. Twenty-nine outpatients (60.42%) and 19 inpatients (39.58%) were included. Hypertriglyceridemia was diagnosed in 16 cases (33.33%), mixed hyperlipidemia in 31 cases (64.58%), and hypercholesterolemia in one case (2.08%). The pretreatment blood lipid concentrations were significantly different after the 59 CMHP treatments (p < .001). The concentrations of TC, TG, HDL-C, and LDL-C decreased significantly after the treatment, and the median ratios of reduction were 67.06% (range: 58.97%-71.87%), 63.33% (range: 55.20%-74.86%), 45.87% (range: 35.86%-52.95%), and 66.09% (range: 44.37%-73.94%), respectively. Three adverse reactions (5.08%) were recorded. No differences were detected in therapeutic parameters, effects, or adverse reactions between the two blood cell separators, there was no difference in Lipoprotein apheresis efficacy. This preliminary study demonstrated the clinical application of CMHP in patients in the treatment of hyperlipidemia. However, further studies are needed applying CMHP with hyperlipidemia.

MTHFR C677T gene polymorphism in patients with coronary heart disease and hypertension treated with enalapril and folic acid: implications for prognosis.

We aimed to investigate the effect of the methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism on the prognosis of patients with coronary heart disease (CHD) and hypertension treated with enalapril and folic acid. A total of 540 CHD patients diagnosed by coronary angiography in our hospital were selected. According to whether there was folic acid intervention, they were divided into a folic acid group, a non-folic acid group and a control group. The genotypes of the MTHFR C677T locus were detected. Hcy concentration and the folate content were determined. In folic acid group, enalapril and folic acid tablets were used to reduce blood pressure, and clopidogrel or ticagrelor were selected according to CYP2C19 genotypes. In non-folic acid group, enalapril tablets were used, and clopidogrel or ticagrelor were selected based on CYP2C19 genotyping. Routine treatment without intervention was used in control group. Patients were prescribed standardized drug treatment and were followed up by an outpatient service or by telephone for 12 months after discharge. We found that the number and proportion of MTHFR C677T gene mutations in the folic acid group, non-folic acid group and control group were 150 (83.3%), 142 (78.9%) and 144 (80.0%), respectively. The recurrence rate of cardiovascular events in the folic acid and non-folic acid groups was significantly lower, and the degree of reduction in the recurrence rate of cardiovascular events in the folic acid and non-folic acid groups was significantly different. The concentrations of TG, TC, and LDL-C in folate and non-folic groups were lower, while HDL-C was higher than that in control group. To sum up, screening high-risk populations with genotypes has great significance in improving the clinical outcome of CHD patients with H-type hypertension. Folic acid supplementation improves the compliance rate of patients' blood pressure levels and improves their clinical prognosis as well.

The effect of metabolic syndrome in patients with type 2 diabetes mellitus: Review

A group of metabolic abnormalities known as the metabolic syndrome (MS) are typified by a number of cardiovascular risk factors that are typically linked to central fat buildup and insulin resistance. Insufficient dietary modifications and weight loss, linked to consistent physical exercise, are regarded as primary and first-choice treatments for multiple sclerosis (MS). These interventions help decrease visceral fat and belly circumference, enhance insulin sensitivity, lower plasma glucose and triglyceride concentrations, increase high-density lipoprotein levels, and ultimately lower the risk factors that lead to type 2 diabetes. The MS is a topic of study in the medical community right now since it is associated with conditions that not only have a high death rate globally but also exhibit rising occurrence. Deregulation of the proteins, fats, and carbohydrates involved in metabolism causes type 2diabetes, which can lead to reduced insulin production, insulin resistance, or a combination of the two. With 90% of cases, type 2 diabetes is the most common of the three kinds of the disease. Being a chronic and complicated illness, diabetes necessitates close medical monitoring in order to lower risks and develop control techniques. Given its rapid epidemic expansion on a global scale in recent years, it is regarded as one of the pandemics of the twenty-first century. This review was non-systematic and advanced, examining the MS in individuals with type 2 diabetes by consulting multiple literatures and analyzing some new publications.

Hepatoprotective Effect of Antrodia camphorata Mycelium Powder on Alcohol-Induced Liver Damage

Background/Objectives: Antrodia camphorata, also known as “Niuchangchih” in Taiwan, is a unique medicinal mushroom native to Taiwan. It is used in traditional medicine to treat various health conditions. In this study, we investigated the efficacy of A. camphorata mycelia on alcohol-induced liver damage, both in vitro and in vivo, in a Good Laboratory Practice (GLP) facility. Methods: The experimental groups consisted of a normal control group (G1), a negative control group (G2), an A. camphorata mycelium powder 50 mg/kg/day administration group (G3), a 100 mg/kg/day administration group (G4), a 200 mg/kg/day administration group (G5), and a positive control silymarin 200 mg/kg/day administration group (G6), with 10 Sprague Dawley rats assigned to each treatment group. Results: We found that treatment with A. camphorata mycelium powder significantly reduced alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, cholesterol, adiponectin, triglyceride, and malondialdehyde concentrations. Histopathological analysis also revealed that the inflammation score significantly decreased in the A. camphorata-treated groups. Conclusion: Based on these results, we conclude that repeated oral administration of A. camphorata mycelium powder is effective in improving alcoholic liver disease.

Effects of artificial diets on lipid and glucose metabolism, antioxidative capacity, and inflammation in the liver of mandarin fish (Siniperca chuatsi)

The mandarin fish (Siniperca chuatsi) is a typical carnivorous fish, which has been able to consume artificial diets after domestication in recent years. However, the potential health consequences of artificial diets in mandarin fish remain unclear. This study aimed to elucidate the molecular mechanisms underlying these concerns. Fish (initial weight: 25.1 ± 0.1 g) were fed with natural (CON group) or artificial diets (AF group) for 8 weeks. Each diet was randomly distributed to sextuplicate circular tanks (300 L) with 40 fish in each tank. The transcriptome analysis revealed significant changes in metabolism-related pathways, particularly those involved in lipid and carbohydrate metabolism. Further investigation confirmed that the artificial diets significantly increased hepatic triglyceride content and fatty acid synthase activity. The artificial diets also significantly increased hepatic glycogen and glucose-6-phosphatase activity. Furthermore, the artificial diets significantly increased hepatic malondialdehyde levels, indicating increased oxidative stress. Antioxidant defense enzyme activities and the expression of antioxidant stress-related genes were significantly decreased. Additionally, the artificial diets significantly increased the expression of proinflammatory genes, including interleukin 1 beta and interferon-gamma. These findings collectively demonstrated that the artificial diets disrupted hepatic lipid and glucose metabolism, leading to oxidative stress and inflammation, thus affecting the health status of mandarin fish.

The effect of Oleoylethanolamide supplementation on lipid profile, fasting blood sugar and dietary habits in obese people: a randomized double-blind placebo-control trial.

Abnormalities in biochemical parameters and changes in eating habits are considered complications of obesity. Oleoylethanolamide (OEA), an endocannabinoid-like compound, has been shown to have protective effects on many metabolic disorders. Given this evidence, the present study aimed to assess the effects of OEA on lipid profile parameters, fasting blood sugar (FBS), and dietary habits in healthy obese people. In this randomized, double-blind, placebo-controlled clinical trial, which was carried out in 2016 in Tabriz, Iran, 60 obese people were enrolled in the study based on inclusion criteria. The intervention group consumed 125mg of OEA capsules, and the placebo group received the same amount of starch twice for 8 weeks. Blood samples (5 mL) were taken at baseline and the end of the study in a fasting state. Serum concentrations of FBS, triglycerides (TGs), high-density lipoprotein cholesterol (HDL-C), and total cholesterol (TC) were measured by enzymatic methods using commercial kits. The low-density lipoprotein cholesterol (LDL-C) concentration was obtained using the Friede-Wald formula. To assess dietary habits, a food frequency questionnaire (147 items) was used at baseline and the end of the study. A value less than < 0.05 was considered to indicate statistical significance. The TG concentration decreased significantly in the intervention group (mean (SD): 166.29 (70.01) mg/dL to 142.22 (48.05) mg/dL, p = 0.047). Changes in the placebo group were not significant (p > 0.05). After adjusting for baseline values and demographic characteristics, the difference in TG between groups remained significant (p = 0.044). Changes in other biochemical parameters were not significant. There was no significant difference between or within groups in terms of food groups. OEA, as a complementary agent, plays a protective role in TG regulation. However, future studies with longer durations are needed to explore the impact of OEA on regulating dietary habits and to identify the mechanisms related to metabolic abnormalities in obese people. The study was registered in the Iranian Registry of Clinical Trials (IRCT) center as IRCT201607132017N30 with URL. www.IRCT.IR in date 03/10/2016.

Palygorskite improves growth performance and prevents liver damage in avian pathogenic Escherichia coli-challenged broiler chickens at an early age.

Avian pathogenic Escherichia coli (APEC) is a major bacterial infection that causes economic losses in the global poultry industry. Palygorskite (PAL) has been shown to enhance growth performance while improving antioxidative and anti-inflammatory properties of broilers. This study evaluated the protective effects of PAL on growth performance and liver function in broilers subjected to APEC challenge. A total of 320 one-day-old male Arbor Acres chicks were divided into four groups with eight replicates of ten birds each, based on a 2×2 factorial arrangement (basal diet or 5g/kg PAL-supplemented diet) and inoculation (bacterial culture medium or APEC). PAL increased body weight gain (BWG) prior to APEC challenge (P < 0.05). However, APEC caused losses in BWG, feed intake (FI), and feed efficiency, along with increased relative hepatic weight, hepatic pathology scores, and hepatic-cell apoptosis rate (P < 0.05). Compared to normal birds, APEC increased interleukin (IL)-1β, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA), and nitric oxide (NO) levels, as well as lysozyme (LZM) and myeloperoxidase (MPO) activities, while decreasing total antioxidant capacity (T-AOC) and IL-10 levels, and total superoxide dismutase (T-SOD) and catalase (CAT) activities in both serum and liver, APEC also raised alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, but reduced total protein (TP), albumin (ALB), immunoglobulin (Ig) A, IgG, and IgM levels in serum (P < 0.05). Moreover, APEC increased hepatic mRNA level of IL-1β, IFN-γ, TNF-α, nuclear factor kappa B, and inducible nitric oxide synthase (iNOS), while inhibited mRNA level of IL-10 (P < 0.05). In contrast, PAL increased BWG and FI, alleviated hepatic-cell apoptosis rate during the challenge period (P < 0.05). An incorporation of PAL reduced triglyceride and NO contents, ALT and AST activities, while increasing TP, ALB, IL-10, IgG, and IgM levels in serum, enhancing serum T-SOD and CAT activities, elevating hepatic T-AOC and CAT activities, inhibiting hepatic MDA accumulation, and reducing IL-1β levels and LZM activity in both liver and serum (P < 0.05). An interactive effect was found for hepatic TNF-α and iNOS mRNA expression, in which PAL inhibited their mRNA expression in APEC-challenged birds (P < 0.05). Overall, PAL addition partially mitigated the negative impact of APEC challenge on growth performance and liver function of broiler chicks at an early age.

7005 Characterization of Clinical Candidate Miricorilant in Preclinical Models for NASH

Abstract Disclosure: T.J. Moll: None. E.M. Viho: None. M. Gentenaar: None. H. Hunt: Employee; Self; Corcept Therapeutics. O.C. Meijer: Grant Recipient; Self; Corcept Therapeutics. J. Kroon: Research Investigator; Self; Corcept Therapeutics. Background and aim: Miricorilant (CORT118335) is a selective glucocorticoid receptor (GR) modulator that is under clinical development for the treatment of non-alcohol steatohepatitis (NASH). It was previously shown that miricorilant treatment effectively lowers hepatic lipid content in mouse models of NASH. The aim of this study is to further characterize the effects of miricorilant in the mouse liver and in human liver tissue. Methods: In mouse liver tissue, we investigated the agonistic and antagonistic properties of miricorilant on GR activity. Eight-week-old male C57BL/6J mice were fed with a control diet or miricorilant-supplemented diet (resulting in an estimated dose of 60 mg/kg/day) for 6 days and 1.5 mg/kg corticosterone was injected to monitor the effects of miricorilant on GR-responsive gene expression. We next accessed the time-dependency of the lipid-lowering effects of miricorilant in the mouse liver. Mice were fed with a high-fat, high-fructose diet (HFHFD) for 3 weeks before treatment with 60 mg/kg/day miricorilant for one, three or six days by oral gavage, and triglyceride content in the liver was measured at endpoint. In human liver tissue, we investigated the effects of miricorilant on precision-cut liver slices (PCLS) obtained from two independent donors with pre-existing steatosis. PCLS were cultured for a total of 168 hours in the presence of 100 nM hydrocortisone, and with or without an exogenous lipid challenge. PCLS were treated with 200-600 nM miricorilant for 96 hours before triglyceride content was measured. RNA-sequencing was performed on PCLS after 24 hours of 200-400 nM miricorilant exposure. Results: In the mouse liver, miricorilant exhibited a unique combination of agonistic and antagonistic properties on GR-responsive gene expression. Miricorilant effectively lowered HFHFD-induced lipid accumulation in the mouse liver as soon as three days after treatment. In human PCLS, miricorilant treatment lowered triglyceride content in both donors. RNA-sequencing revealed respectively 40 and 128 differentially expressed genes after 200 and 400 nM miricorilant treatment. Pathway analysis revealed very-low-density lipoprotein (VLDL) particle remodeling as a significant enriched pathway, providing a possible explanation for the lowered triglyceride content upon miricorilant treatment. Conclusion: Miricorilant rapidly and effectively lowers hepatic lipid content in the mouse liver and in human PCLS, possibly via modulation of VLDL remodeling. Presentation: 6/2/2024

Use of corn germ meal and bile acids in laying quail diets: effect on production, reproduction, health and economics

In the present study, the effects of corn germ meal (CGM) and bile acids (BA) inclusion in Japanese quail diet on the productive and reproductive performance, egg quality, and serum biochemical parameters were evaluated. Six wk old, 480 Japanese quail birds were randomly divided into 6 groups (60 hens and 20 males / group) with 4 replicates / group. CGM was incorporated at 0, 10 and 20 % of diet for groups (G1, G2, and G3), while G4, G5 and G6 had the same levels of CGM with BA addition (500 g/ton feed). The egg production (EP), egg mass (EM) and feed intake (FI) were significantly reduced (P < 0.05) in response to CGM feeding which was clear with the 20 %. CGM at 10% inclusion enhanced the feed conversion ratio (FCR), while incorporation at 20% deteriorated (P < 0.05) it compared to the control group. Addition of BA to the CGM-containing diets reduced the FI, improved FCR, and ameliorated the negative effects on EP, EM and FCR in the CGM fed birds. Feeding on 10% CGM single or combined with BA increased fertility %, chick's weight after hatching and reduced embryonic death. Serum concentrations of triglyceride and low-density lipoprotein (LDL) were significantly increased (P < 0.05) with CGM inclusion. Dietary CGM inclusion without or with BA was associated with higher levels of serum total cholesterol and low-density lipoprotein when compared to their control groups. BA addition to the CGM containing diets significantly reduced the alanine transaminase (ALT) activity (P ˂ 0.05) compared to their corresponding groups fed BA free diets especially those received the 10% CGM. Birds fed on 10% CGM supplemented with BA showed the highest economic efficiency values. In conclusion, CGM inclusion at 10% with BA supplementation could be used as alternative ingredient in the Japanese quail layer diet without compromising their productive performance.

12248 Leptin Signaling Deficiency At The Central Nervous System Reduces Hepatic Glucagon Sensibility Disturbing Liver Amino Acid Metabolism Leading To Hyperaminoacidemia And Hyperglucagonemia In Male Adult Rats

Abstract Disclosure: C. Pintado Losa: None. L. Mazuecos Fernández-Pacheco: None. Ó. Gómez Torres: None. S. Artigas Jerónimo: None. E. Burgos Ramos: None. I. Ramos: None. J. Poveda Colado: None. C. Arribas Mocoroa: None. A. Andres: None. N. Gallardo: None. Leptin regulates glucagon secretion, but the contribution of central leptin resistance to circulating glucagon levels and hepatic glucagon signaling is poorly studied. To analyze the effects of deficient hypothalamic leptin signaling on glucagon levels and hepatic glucagon signaling, we infused intracerebroventricularly a rat-specific leptin receptor antagonist SLA (0,2 µg/day) for 21 days to male 3-month-old Wistar rats (n=12) or PBS (n=12). At day 21 of treatment and 30 min before sacrifice, a glucagon challenge (100 µg/kg) was performed in 16h fasted rats. After sacrifice, blood and liver samples were collected for analysis. Serum hormones, urea, and total and branched-chain amino acids (BCAA) levels were determined using commercial kits and by RP-HPLC. Hepatic neutral lipid was extracted and quantified using an enzymatic kit and the hepatic content of mRNA and proteins involved in lipid and amino acid metabolism were measured by RT-PCR and Western-Blot. Additionally, the HOMA-IR and glucagon-alanine index were determined in all rats. Blocking central leptin signaling led to impaired fat oxidation and triglyceride export capacity in the liver, resulting in a 50% increase in hepatic triglyceride content in SLA-treated rats. Glucagonemia increased in SLA-treated rats, indicating impaired hepatic glucagon sensitivity. In fact, in response to exogenous glucagon, the hepatic phosphorylation of CREB was reduced by 55% in SLA-infused compared to control rats. Importantly, SLA treatment significantly decreases the expression and the activity of BCKDH (branched-chain alpha-keto acid dehydrogenase) in the liver, leading to a reduction in the BCAA degradation in SLA-treated rats, hyperaminoacidemia, and decreased serum urea levels. In conclusion, deficient hypothalamic leptin signaling alters glucose, lipid, and amino acid homeostasis, highlighting the relevance of central leptin action in the control of peripheral metabolism and glucagon sensitivity. Funding: Work supported by Ministerio de Ciencia, Innovación y Universidades under project grants RTI2018-098643-B-I00, PID2021-128243OB-I00, and from the University of Castilla-La Mancha and the European Regional Development Fund (FEDER) under project grant 2023- GRIN-34280. Presentation: 6/1/2024

The influence and mechanism of fish collagen peptide and egg yolk lecithin on proliferation and lipid composition in feline adipocytes.

The influences of fish collagen peptide (FCP) and egg yolk lecithin (EYL) on the proliferation, fat accumulation and triglyceride content in feline adipocytes were investigated in this work, aiming at unveiling the mechanism of fat accumulation for cheek of feline animals. The lipogenic changes of adipocytes in the presence of FCP and EYL were determined by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The results demonstrated that FCP of 10mg/mL had the strongest cell activity, with a relative increment rate of 156 ± 0.23%, and the triglyceride content reached 215.9 ± 3.86 mmol/L. By comparison, it was observed that an EYL concentration of 5mg/mL elicited the highest cell activity, exhibiting a relative increment rate of 152 ± 0.60%, and the level of triglyceride content was noted to reached 256.56 ± 25.68 mmol/L. After the feline adipocytes were treated with different concentrations of two active substances, fat formation and lipid droplets were found by oil red O staining. Liposome analyses confirmed that the formation of lipid compounds was regulated by FCP and EYL through pathways involved in lipid metabolism, notably including inositol phosphate insulin resistance, and phosphatidylinositol signaling pathways. This regulation was found to enhance cell vitality and facilitate fat accumulation. These findings provide a new strategy for the development of nutritional and healthy products or foods that promote feline cheek.

Changes in Selected Biochemical Markers of Honey Bees Exposed to Fermented Common Tansy Solution (Tanacetum vulgare L.).

Honey bees use pollen and nectar from flowers to produce food. Because they often forage on crops, they are at risk of being exposed to plant protection products (PPPs), both directly and in stored food. Due to the adverse effects of synthetic PPPs on pollinators, biopesticides may be a viable alternative. Common tansy extract is used as one of the natural substitutes for synthetic pesticides. In our study, the effect of fermented common tansy extract on aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma-glutamyl transpeptidase (GGTP) activity and the concentration of triglycerides (TGs), total protein (TP), total antioxidant status (TAS), and glucose in honey bee workers' hemolymph was assessed. These biochemical markers give valuable information about the immunity, detoxification, and nutrition of a bee's body. Caged bees were given tansy extract added at various concentrations in sugar syrup for 24 h. Then, they were provided with only sugar syrup. After 7 days of the experiment, hemolymph was collected and analyzed. We observed changes in the activity of AST, ALT, GGTP enzymes and TG, TP, and glucose levels, but not all changes were statistically significant. In terms of AST activity, statistically significant differences were found. All groups tested, including the negative control group, showed reduced enzyme activity values compared to the positive control group. In TG concentration, differences were observed between the groups receiving 2% extract and 1% ethanol. Glucose levels differed between the groups receiving 1% extract and 2% extract and between the positive control group and 1% extract. Bee body proper functioning is affected by changes in enzyme activity, especially those responsible for immunity and detoxification, such as AST, ALT, ALP, and GGTP. Despite the short time of bees' exposure to the agent, the results of study show visible effects. Our results provide a basis for further research on the impact of tansy extract on honey bees.

Resveratrol inhibits white adipose deposition by the ESR1-mediated PI3K/AKT signaling pathway

Excessive adipose accumulation is the primary cause of obesity. Resveratrol (RES), a natural polyphenolic compound, has garnered significant attention for its anti-obesity properties. However, the precise mechanisms by which RES influences fat deposition have not yet been explored. In this study, the aim was to identify the target proteins and associated pathways of RES in order to elucidate the mechanisms by which RES reduces fat deposition. In this study, mice were administered 400 mg/kg of RES via gavage for 12 weeks. We found that while 400 mg/kg RES had no impact on the growth of the mice, it significantly reduced the weight of various white adipose tissues, as well as the serum and liver concentrations of total cholesterol and triglycerides. Network pharmacology identified 15 potential targets of RES and highlighted the PI3K/AKT signaling pathway as a key pathway. Molecular docking and dynamic simulations suggested that ESR1 might be the target protein through which RES exerts its anti-fat deposition effects. In vitro experiments revealed that ESR1 promotes the proliferation and inhibits the differentiation of 3 T3-L1 adipocytes, and suppresses the PI3K/AKT signaling pathway. Silencing the ESR1 gene altered the ability of RES to inhibit cell differentiation via the PI3K/AKT pathway. Gene expression results in subcutaneous adipose tissue, epididymal fat tissue, and liver tissue of mice were consistent with observations in cells. In summary, RES reduces white fat deposition by directly targeting the ESR1 protein and inhibiting the PI3K/AKT signaling pathway. Our findings provide new insights into the potential use of RES in the prevention and treatment of obesity.

Effect of feeding two types of concentrates in morning and evening meals and two types of fat supplement on diurnal patterns of plasma parameters in lactation dairy cows

The diurnal patterns of feed intake and TMR composition influence blood parameters in dairy cows; however, the effect of feeding TMR with different compositions in the morning and evening meals is not well characterized. In a completely random design, forty Holstein cows (110 ± 30 days postpartum) were randomly assigned to four treatment groups, with 10 cows per treatment. The treatments were as follows: a base diet containing 50 % cereal starch from barley grain and 50 % cereal starch from corn grain in the morning and evening meal + either prilled fatty acids supplement (EqP) or calcium salts of fatty acids supplement (EqCS); a base diet containing 25 % cereal starch from barley grain, 75 % cereal starch from corn grain in the morning meal, and 75 % cereal starch from barley grain and 25 % cereal starch from corn grain in the evening meal + either prilled fatty acids supplement (DiP) or calcium salts of fatty acids supplement (DiCS). Dry matter intake was affected by treatments (P ˂ 0.01). The highest intake was observed for EqP, EqCS, DiP,and DiCS. 3.5 % fat-corrected milk and milk fat percentage was significantly higher for EqP than other treatments (P ˂ 0.01), but other milk components were not significantly different among treatments (P ˃ 0.05). A significant difference was observed for glucose, cholesterol, TG, AST, and insulin concentration among treatments (P ˂ 0.01). The concentrations of all plasma parameters (glucose, cholesterol, TG, BUN, AST, insulin, and NEFA) significantly changed over 24-h period (P ˂ 0.01). Plasma concentrations of glucose, cholesterol, TG, BUN, AST, insulin, and NEFA displayed a treatment-by-time interaction (P ˂ 0.01). In general, changes in the cereal starch ratio (corn and barley) in TMR in each meal during a day affect the 24-h concentration of plasma parameters, but there was no strong evidence of leverage for cows’ metabolism.

A-226 Validation of Calculated Glycerol Corrected Triglycerides

Abstract Background Patients with elevated free glycerol can have falsely elevated triglyceride concentrations known as pseudohypertriglyceridemia. Most clinical laboratory triglycerides methods measure the concentration of glycerol released following the enzymatic hydrolysis of triglycerides to quantitate triglycerides. Thus, increased free glycerol can lead to falsely increased triglyceride concentration. Identifying and measuring the interference is necessary to effectively manage patient care interventions. A glycerol-corrected triglyceride assay was discontinued from Roche Diagnostics in 2021. We created a process to determine glycerol-corrected triglycerides by measuring glycerol directly using an assay from Randox Laboratories. Methods Glycerol (Randox Laboratories) and nonglycerol-blanked triglycerides (Roche Diagnostics) were measured in serum using the Roche Cobas c501. Stability (n=10) and expected values (n=206) were determined using serum from healthy volunteers. Linearity (n=5) and precision (5 replicates on 2 runs over 5 days) were determined using residual clinical serum spiked with exogenous glycerol as necessary to span the analytical measuring range. Glycerol corrected triglyceride concentrations were calculated by subtracting glycerol (mmol/L) from triglyceride (mmol/L) and converting to mg/dL (x88.59). Samples previously measured using the Triglyceride/Glycerol Blanked Reagent (n=53, Roche Diagnostics) were used to assess accuracy. (Figure 1) Results Glycerol measures were stable 3 days refrigerated and 30 days frozen. Precision coefficient of variation was 1.0% within run and 7.1% within lab. Expected values were 10 - 109 mcmol/L, and the method was linear between 2 - 3,971 mcmol/L. The average bias between methods was 11.0% with slope of 1.02 and R2&amp;gt;0.97 (Figure 1). Conclusions Glycerol can be accurately measured using the Randox method and the result can be used to generate an accurate glycerol-corrected triglyceride concentration.

B-310 Freezing and thawing serum without mixing prior to analysis causes false low Thyroglobulin results

Abstract Background Thyroglobulin (Tg), a glycoprotein that is synthesized in follicular cells of the thyroid gland, is the precursor of Thyroxin (T4) and Triiodothyronine (T3). Tg is a biomarker used for many clinical conditions, including monitoring for recurrence in patients with differentiated thyroid carcinoma. Typically, Tg levels drop to very low or undetectable levels after surgical excision and/or radioiodine ablation of the thyroid, with post-operative levels greater than 1-2 ng/ml indicating potential disease recurrence. Discordant Tg results in a patient sample were identified during a quality check of Tg results. This study investigates the root of cause of the discrepant Tg results. Given that: 1) Tg analyses were performed twice a week at our laboratory due to low daily test volume and patients’ samples received on an unanalytical day were stored at -20°C until analysis; 2) Tg analyses were performed by various medical technologists; 3) sample mixation prior to analysis was not routine practice for our automated laboratory, we hypothesized that the discrepancy was due to inconsistent sample handling, e.g., mixation vs unmixation prior to analysis. Methods Analysis of the daily QC and PT results in the period of the incident occurence ruled out causes in analytical phase. We split 52 de-identified patient serum samples into two equal aliquots and subsequently stored both aliquots at -20°C for 48 hours. Both aliquots of each sample were thawed at room temperature for 1 hour. One of the aliquots was analyzed for Tg without mixing, and the other was analyzed after mixing using a Vortexer for 4 seconds at 250 xg. To investigate if Tg concentrations form a gradient from the top to the bottom of a red-top vacutainer tube, five patient samples with Tg at 2.59, 2.79, 2.92, 52.37 and 144.12 ng/mL were thawed without mixing following storage at -20°C for 48 hours. Five layer (600 μl/layer) aliquots were carefully and sequentially taken from top to bottom for each specimen, and the Tg concentration in the aliquot of each layer was determined using the Beckman Coulter Access-2 Immunoassay. Results Of the 52 patients samples, 14 Tg results were excluded from statistical analysis due to exceeding the assay dynamic range (2 above and 12 below the upper and low limits, respectively). Analysis of the remaining 38 patient samples showed the mean of Tg concentrations was 22.8% (SD 18%) higher in mixed samples than the un-mixed samples. Regression analysis showed an R2 = 0.9976 (Mixed Tg = 1.1523 x Un-mixed + 0.4174, range 0.1 - 275.7 ng/mL, intercept 95%CI -0.721 to 1.556, slop 95%CI 1.1331 to 1.1715, p-value &amp;lt;0.001). Results of Tg gradient analysis showed significant higher Tg values in the 2nd, 3rd, 4th and 5th than the 1st layer by 26% (SD 11%), 63% (26%), 112% (57%), and 160% (76%), respectively. Conclusions False Tg values were found in samples without mixing before analysis after thawing from frozen state, due to formation of Tg concentration gradient in test tube. For obtaining reliable results, samples must be mixed before analysis when thawed from frozen storage.

A-119 Lipemia Interference

Abstract Background Lipemic specimens is a common problem in clinical laboratories. Analytical results may be perturbed by lipemia, leading to misdiagnosis and unnecessary treatments for patients. Serum index testing is an important preanalytical test for assessing the potential effect of endogenous HIL interferents (hemoglobin (H), bilirubin (I) or lipids (L)) on test results. Automated HIL systems are not standardized and differ in their response. HIL induced interference should be verified by all clinical laboratories. Intralipid, a commercial lipid emulsion with particle size ranges from 200 to 600 nm is often used to verify the L index. However, intralipid lacks particles that mimic large VLDL, as well as the lower and upper ranges for chylomicrons size. In contrast, patient samples contain a complex mixture of macromolecular lipid and protein structures. Therefore, lipemia induced by Intralipid is not identical to lipemia in patient serum samples. The purpose of this study is to increase awareness of lipid interference and demonstrate the relationship of a lipid L Index to intralipid and triglyceride interference. Methods A set of 16 panels were prepared from a commercially available Triglyceride rich Lipoprotein material. These panels spanned a lipid concentration from 0 to 3000 mg/dL. Panels were tested on the Alinity c and ARCHITECT c8000 TRIG2 assays (04U0615 and 04T1016 respectively) on 2 reagent lots in a single run to determine the Triglyceride concentration. Then these samples were assessed for an HIL L score using the Abbott HIL saline protocol. Results The relationship between the Triglyceride rich Lipoprotein and the L Index was linear with a high correlation (r= 0.999) for Alinity and ARCHITECT respectively. The L Index underestimated the Triglyceride Rich Lipoprotein concentration. The reason for the lower estimate of Triglyceride rich lipoprotein specimens on an L Index is likely due to the differences in size of the lipid particles between the two artificial lipid materials, Intralipid and Triglyceride rich lipoprotein. In addition, while the triglyceride assay provides a quantitative measurement of the lipemic concentration, the L Index is designed to provide an estimated result. Conclusions Intralipid is an acceptable material used to verify spectrophotometric HIL systems. The L Index provides an estimate of sample turbidity, not the concentration of Triglyceride. Vendor purchased human derived Triglyceride rich lipoprotein, which contains a heterological mixture of lipid particles, may be regarded as a more appropriate material for assessing lipid interference. Thus, while recovery of the L Index correlates well to the Intralipid concentration, it can have a poorer correlation with vendor purchased samples containing Triglyceride due to the different vesicle sizes in these materials. This study demonstrates the Triglyceride rich material used to assess lipid interference in Sigma Strong assays provides a difference in response between the Triglyceride concentration of this material determined on a Triglyceride assay vs the L Index. The choice of lipemic material to use, (eg. artificial, native), can influence the results. Thus, care should be taken in comparing results for these materials.

A-228 Evaluation of an Equation for Low-Density Lipoproteins-Triglycerides in Predicting ASCVD Risk in the UK Biobank Population

Abstract Background Low-density lipoprotein-cholesterol (LDL-C) is the main univariate risk marker recommended for initial atherosclerotic cardiovascular disease (ASCVD) risk stratification. LDL, however, transports other constituents that can serve as pro-atherogenic risk markers. Like LDL-C, the triglyceride content of LDL (LDL-TG) can also be estimated from the lipid panel. Using data from UK Biobank (UKB), we compared LDL-C to LDL-TG for predicting future ASCVD events. Methods A UKB dataset (N=270,436) of primary prevention individuals not on lipid lowering therapy was used for analysis. LDL-C was calculated by the following equations: Friedewald, Extended Martin-Hopkins, Sampson-NIH, and Enhanced Sampson-NIH. LDL-TG was calculated by our newly devised equation (eLDL-TG = TG/38.46 + nonHDL-C/5.75 + (TG/nonHDL-C)/0.103 + (1/HDL-C)/0.0041 - 2.95)), which was derived from a dyslipidemic cohort (N=40,202) analyzed by the beta-quantification reference method. Results Although there was only a modest comparison between eLDL-TG and measured LDL-TG (R2=0.601, slope=0.611), eLDL-TG appeared to be a better risk marker than LDL-C in UKB. When the population was divided into quintiles and analyzed by survival curve analysis, eLDL-TG was better than LDL-C in segregating patients into low- and high-risk groups (Figure). Similar findings on the relative value of LDL-TG over other risk markers were also found when analyzed by ROC analysis (AUC: eLDL-TG&amp;gt;HDL-C&amp;gt;remnant-cholesterol&amp;gt;NonHDL-C&amp;gt;TG&amp;gt;apoB&amp;gt;Sampson-NIH-LDL-C). By Cox-proportional hazard analysis eLDL-TG showed alongside apoB and NonHDL-C the strongest association with future ASCVD events. Discordance analysis showed that when stratified as LDL-TG/LDL-C as High/Low there was an 26% increased risk for ASCVD, and when as High/High showed an 40% increased risk for ASCVD. Conclusions Like LDL-C, LDL-TG levels can also be estimated from the standard lipid panel. If our findings hold in other populations, the superiority of LDL-TG over LDL-C as a risk marker suggest that the reporting of calculated LDL-TG along with LDL-C could be a new strategy for improving ASCVD risk stratification.

B-167 Quantification of Thyroglobulin in Serum for Clinical Research using SISCAPA Workflow Combined with LC-MS/MS

Abstract Background Accurate measurement of Thyroglobulin (Tg) using existing immunoassay-based techniques can be challenging in the presence of anti-Tg antibodies (TgAb), which can prevent the binding of Tg to assay antibodies thus leading to non-quantifiable Tg concentrations. The Stable Isotope Standards and Capture with Anti-Peptide Antibodies (SISCAPA™) Workflow combined with LC-MS/MS has been successfully employed to circumvent this problem by digesting the serum sample thus eliminating interfering TgAb and measuring a Tg-specific surrogate peptide instead. Methods Using a Andrew+™ Pipetting Robot, denaturation and reduction buffer was added to 250µL serum samples and incubated at 37°C. Internal standard and trypsin were added and samples incubated at 37°C. The samples were quenched with protease inhibitor and Anti-Tg FSP mAb SISCAPA was added to the serum samples and mixed for 60 minutes. Using a magnetic array plate, the beads were left to pull down, samples were discarded, and the beads were twice washed with PBS/CHAPS prior to elution with acetonitrile with formic acid. Using an ACQUITY™ UPLC™ I-Class PLUS FL System, samples were injected onto a Waters™ XSelect™ HSS T3 Column using a water/acetonitrile/formic acid gradient elution profile and the Tg FSP peptide was quantified using a Waters Xevo™ TQ Absolute Mass Spectrometer. Results The method demonstrated no significant carryover or matrix effects and was shown to be linear from 0.1 - 50 ng/mL. Analytical sensitivity investigations indicate the analytical sensitivity of this method would allow precise quantification (&amp;lt;20%) at 0.1 ng/mL with S/N (PtP) &amp;gt;10:1. Coefficients of variation (CV) for total precision and repeatability on 5 analytical runs for low, mid and high QCs were all &amp;lt; 9.5% (n = 25) for both automated and manual precision assessments. Comparison with samples from the UK NEQAS scheme demonstrated significant method bias of -40% for the developed LC-MS/MS method. Re-assignment of the calibrator concentrations using the EQA samples reduced the bias to -5.5%, indicating differences in the calibration materials used for these measurements. Conclusions A LC-MS/MS clinical research method for serum thyroglobulin was developed using SISCAPA, followed by analysis using ACQUITY UPLC I-Class PLUS FL System and the Xevo TQ Absolute Mass Spectrometer. The method provides analytical sensitivity down to 0.1 ng/mL from 250 µL serum, while providing sufficient sample for re-analysis. The method demonstrates excellent linearity across the calibration range, with no significant carryover, interferences, and matrix effects. Total reproducibility and repeatability of the method was ≤9.5% RSD for manual and automated sample preparation, using the Andrew+ Pipetting Robot. In addition, the Andrew+ Pipetting Robot can minimize user touch-time, allowing a full plate to be prepared within four hours without user intervention.