Triglyceride Removal Research Articles

Extraction of Coumarin Mixture from Tamanu Oil using Food-Grade

Indonesia’s flora and fauna are divided into four significant ecosystems: freshwater (rivers and lakes), marine, semi-terrestrial (mangrove and riparian), and terrestrial. Tamanu (Calophyllum inophyllum) is a member of the mangrove-associated plant species belonging to the mangosteen family. It is also known as Alexandrian laurel, bintangur (Malaysia), Borneo-mahogany, polanga (India), and nyamplung (Indonesia). The tamanu seed can produce oil with a 60-75% yield. It is applicable to treat various skin problems, including eczema, psoriasis, herpes, acne, haemorrhoids, and certain allergies. It is also beneficial as an antimicrobial, anticancer, anti-HIV, anti-inflammation, and anticoagulant agent. This is because tamanu oil contains bioactive compounds named coumarin. Previous works isolated coumarins with toxic solvents and using methods that are hard to scale up. Therefore, this study aims to know the best concentration of food-grade ethanol, the number of stages, and the solvent system to extract crude coumarin mixtures from tamanu oil by batchwise solvent extraction. The free fatty acid and triglyceride removal was also studied. The initial contents of free fatty acid, coumarin mixture, and triglyceride in tamanu oil were 25.87%, 11.96%, and 51.16%, respectively. Tamanu oil was extracted simultaneously with binary solvents (n-hexane and 70%/80%/90% aqueous food-grade ethanol) and a single solvent (96% food-grade ethanol). Each fraction was analysed by Thin Layer Chromatography (TLC) and High Temperature-Gas Chromatography (HT-GC). The best result was obtained using 80% aqueous food-grade ethanol, eight extraction stages, and a binary solvent system, which produced 48.64±6.57% purity and 88.68±1.38% recovery of coumarin mixture. Moreover, it was found that the method removed 64.15% of free fatty acid and 99.96% of triglyceride from the crude tamanu oil after eight stages of batchwise extraction.

Adipose Tissue Dynamics: Cellular and Lipid Turnover in Health and Disease.

The alarming increase in obesity and its related metabolic health complications, such as type 2 diabetes, has evolved into a global pandemic. Obesity is mainly characterized by excessive accumulation of adipose tissue, primarily due to an imbalance between energy intake and expenditure. Prolonged positive energy balance leads to the expansion of existing adipocytes (hypertrophy) and/or an increase in preadipocyte and adipocyte number (hyperplasia) to accommodate excess energy intake. However, obesity is not solely defined by increases in adipocyte size and number. The turnover of adipose tissue cells also plays a crucial role in the development and progression of obesity. Cell turnover encompasses the processes of cell proliferation, differentiation, and apoptosis, which collectively regulate the overall cell population within adipose tissue. Lipid turnover represents another critical factor that influences how adipose tissue stores and releases energy. Our understanding of adipose tissue lipid turnover in humans remains limited due to the slow rate of turnover and methodological constraints. Nonetheless, disturbances in lipid metabolism are strongly associated with altered adipose tissue lipid turnover. In obesity, there is a decreased rate of triglyceride removal (lipolysis followed by oxidation), leading to the accumulation of triglycerides over time. This review provides a comprehensive summary of findings from both in vitro and in vivo methods used to study the turnover of adipose cells and lipids in metabolic health and disease. Understanding the mechanisms underlying cellular and lipid turnover in obesity is essential for developing strategies to mitigate the adverse effects of excess adiposity.

Effects of a genetic variant rs13266634 in the zinc transporter 8 gene (SLC30A8) on insulin and lipid levels before and after a high-fat mixed macronutrient tolerance test in U.S. adults

BackgroundThe common C-allele of rs13266634 (c.973C>T or p.Arg325Trp) in SLC30A8 (ZNT8) is associated with increased risk of type 2 diabetes. While previous studies have examined the correlation of the variant with insulin and glucose metabolism, the effects of this variant on insulin and lipid responses after a lipid challenge in humans remain elusive. The goal of this study was to determine whether the C-allele had an impact on an individual’s risk to metabolic syndromes in U.S. adults. MethodWe studied the genotypes of rs13266634 in 349 individuals aged between 18 and 65 y with BMI ranging from 18.5 to 45 kg/m2. The subjects were evaluated for insulin, glucose, HbA1c, ghrelin, and lipid profiles before and after a high-fat mixed macronutrient tolerance test (MMTT). ResultsWe found that the effects of variants rs13266634 on glucose and lipid metabolism were sex-dimorphic, greater impact on males than on females. Insulin incremental area under the curve (AUC) after MMTT was significantly decreased in men with the CC genotype (p < 0.05). Men with the CC genotype also had the lowest fasting non-esterified fatty acid (NEFA) concentrations. On the other hand, the TT genotype was associated with a slower triglyceride removal from the circulation in men after MMTT. The reduced triglyceride removal was also observed in subjects with BMI ≥ 30 carrying either the heterozygous or homozygous T-allele. Nevertheless, the SNP had little effect on fasting or postprandial blood glucose and cholesterol concentrations. ConclusionWe conclude that the CC genotype negatively affects insulin response after MMTT while the T-allele may negatively influence lipolysis during fasting and postprandial blood triglyceride removal in men and obese subjects, a novel finding in this study.

Insight into triglyceride-reducing potential of quercetin in blood plasma environment

Nowadays, hyperlipidemia and abnormal triglyceride levels are global problems menacing human health worldwide. The objective of this study is in-vitro reduction of plasma triglyceride level of the human blood using quercetin through batch adsorption, process optimization, and enzyme-based photometric investigations. The molecular dynamic simulation was also employed to study the peroxidase inhibition effects of quercetin through the enzymatic assay. The best response to the objective function (final triglyceride concentration in the range of 120–170 mg/dl) was obtained at the optimal conditions i.e., contact time of 63 ± 2 min, shaking rate of 264 ± 0.2 rpm and adsorbent dose of 0.037 ± 0.001 g, using experimental design, response surface methodology. Optimization results indicated that quercetin dosage is the most important variable affecting triglyceride removal. Various adsorption isotherms and kinetic models were evaluated to describe the adsorption process. Langmuir type II and Freundlich isotherms as well as pseudo-second-order kinetic model fitted the experimental data appropriately. Based on inhibitory studies and molecular dynamic simulations, quercetin might adsorb plasma triglycerides and inhibit peroxidase enzyme.

Easily operated COF-based monolithic sponges as matrix clean-up materials for non-targeted analysis of chemical hazards in oil-rich foods

Non-targeted analysis of chemical hazards in foods plays a crucial role in controlling food safety. However, because it brings forward high demand for sample pretreatment, materials suitable for the pretreatment of foods, especially animal foods, are rare. Herein, covalent organic frameworks (COF)-based monolithic materials were constructed by three successive steps: preparation of polydimethylsiloxane (PDMS) sponge using sugar cube as a sacrificial template, loading of a heteroporous COF on PDMS sponge via ultrasonic or in-situ growth method, coating of the obtained PDMS@COF by polydopamine (PDA) network. As-prepared PDMS@COF@PDA sponges were demonstrated to work well in sample pretreatment of animal foods for non-targeted analysis of chemical hazards. After a simple vortex treatment for about 2 min, more than 98% triglycerides, the main interfering matrix components in animal foods, could be removed from lard and pork samples, accompanied by “full recovery” (recovery efficiencies: ≥63%) of 44 chemical hazards with different physicochemical properties. Besides providing promising sample pretreatment materials for non-targeted food safety analysis, this work also paves a feasible way to improve COF-based monolithic materials and thus promote their practical applications, because we found that the introduction of PDA network on COF-based monolithic material surface could play a role in “killing three birds with one stone”: enhancing the stability of the materials by overcoming the detachment of COF during operations; controllably adjusting hydrophobic and hydrogen-bonding interactions on the material surface to promote the removal of triglycerides; weakening the hydrophobic and π-π interactions between COF and chemical hazards to increase the recoveries of chemical hazards.

Studies on the partial characterization of extracted glycosaminoglycans from fish waste and its potentiality in modulating obesity through in-vitro and in-vivo.

Glycosaminoglycans (GAGs) are bioactive polysaccharides or glycoconjugates found in the fish waste having significant health impacts. In the present study it has been attempted to extract GAGs from mackerel fish waste through chemical and enzymatic methods. Further, the extracted GAGs (e-GAGs) were analyzed for their composition (uronic acid, total sugar & sulfate), chemical characterization was carried out through techniques of scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) & Proton NMR. Further, probable major GAGs present was identified by enzymatic digestion. The biological potential of the extracted glycoconjugate was assessed further through in-vitro and in-vivo studies. In-vitro biological activity showed good lipase inhibition (IC50, 2.6mg/mL) and bile acid binding properties (dose-dependent). Lipid accumulation lowered in the e-GAGs differentiated 3T3L1 preadipocyte cells have also been observed. The high fat fed animal (in-vivo) study showed ameliorative effect via reducing blood sugar∼1.28↓, lipid profile↓, plasma insulin∼3.5↓, improved glucose tolerance, and homeostatic model assessment for insulin resistance (HOMA-IR, ∼3.0↓). Furthermore, elimination of bile acid (BA) due to GAG-BA binding properties resultant in removal of elevated fecal triglyceride and cholesterol suggesting its lipid lowering activity. Regulation of various proteins linked to carbohydrate and lipid metabolism including fatty acid synthase (FAS), low density lipoproteins receptor (LDL-R), 7α-hydroxylase, glucose transporter-4 (GLUT4) and Peroxisome proliferator- activated receptor gamma (PPAR-γ) were significant (p < 0.05) with e-GAGs treatment when compared to HFD group. Thus, the e-GAGs showed potential hypolipidemic activity through elimination of bile acid binding property together with regulating the specific protein related to obesity and its associated complications.

Rapid quantification of fatty acids in plant oils and biological samples by LC-MS

Analysis of fatty acids (FA) in food and biological samples such as blood is indispensable in modern life sciences. We developed a rapid, sensitive and comprehensive method for the quantification of 41 saturated and unsaturated fatty acids by means of LC-MS. Optimized chromatographic separation of isobaric analytes was carried out on a C8 reversed phase analytical column (100 × 2.1 mm, 2.6 μm core–shell particle) with a total run time of 15 min with back pressure lower than 300 bar. On an old triple quadrupole instrument (3200, AB Sciex), pseudo selected reaction monitoring mode was used for quantification of the poorly fragmenting FA, yielding limits of detection of 5–100 nM. Sample preparation was carried out by removal of phospholipids and triglycerides by solid-phase extraction (non-esterified fatty acids in oils) or saponification in iso-propanol (fatty acyls). This is not only a rapid strategy for quantification of fatty acyls, but allows the direct combination with the LC-MS-based analysis of fatty acid oxidation products (eicosanoids and other oxylipins) from the same sample. The concentrations of fatty acyls determined by means of LC-MS were consistent with those from GC-FID analysis demonstrating the accuracy of the developed method. Moreover, the method shows high precisions with a low intra-day (≤ 10% for almost all fatty acids in plasma and ≤ 15% in oils) and inter-day as well as inter-operator variability (< 20%). The method was successfully applied on human plasma and edible oils. The possibility to quantify non-esterified fatty acids in samples containing an excess of triacylglycerols and phospholipids is a major strength of the described approach allowing to gain new insights in the composition of biological samples.Graphical abstract

Human apoCIII transgenic mice with epicardial adipose tissue inflammation and PRESERVATION of the cardiac plexus.

Hypertriglyceridemia is a result of the increase in the serum levels of lipoproteins, which are responsible for the transport of triglycerides and can be caused by genetic and/or metabolic factors. Animal models which either express or lack genes related to changes in the lipoproteins profile are useful to understand lipid metabolism. Apolipoprotein CIII (apoCIII) is an important modulator of hepatic production and peripheral removal of triglycerides. Mice that overexpress the apoCIII gene become hypertriglyceridemic, showing high concentrations of free fatty acids in the blood. Since hypertriglyceridemia is related to atherosclerosis, and the latter refers to cardiac alterations, this study aimed at evaluating the morphological, morphometric and quantitative profiles of the cardiac plexus, as well as the morphometric and histopathological aspects of the epicardial adipose tissue in human apoCIII transgenic mice. Therefore, 8-12-month-old male C57BL/6 mice that overexpressed human apoCIII (CIII) and their respective controls were used. Our results showed that overexpression of human apoCIII did not modify morphological or quantitative parameters of cardiac plexus neurons; however, age increased both, the area and the number of such cells. Furthermore, there was a direct correlation of this dyslipidemia to the thickening of periganglionar type 1 collagens. On the other hand, this overexpression caused epicardial adipose tissue inflammation and an increase in the area of the adipocytes, thus, favoring the recruitment of inflammatory cells in this tissue. In conclusion, this overexpression is harmful since it is related to an increase in cardiac adiposity, as well as to a predisposition to an inflammatory environment in the epicardial fat and to the incidence of cardiovascular diseases.

Severe L-asparaginase-induced Hypertriglyceridaemia Treated with Plasmapheresis.

Pancreatitis following L-asparaginase-induced hypertriglyceridaemia is a severe and potentially reversible condition in both adults and children treated for acute lymphoblastic leukaemia.Early plasmapheresis is more effective to rapidly decrease serum triglycerides.

A Comprehensive Update on the Chylomicronemia Syndrome.

The chylomicronemia syndrome is characterized by severe hypertriglyceridemia and fasting chylomicronemia and predisposes affected individuals to acute pancreatitis. When due to very rare monogenic mutations in the genes encoding the enzyme, lipoprotein lipase, or its regulators, APOC2, APOA5, GPIHBP1, and LMF1, it is referred to as the familial chylomicronemia syndrome. Much more frequently, the chylomicronemia syndrome results from a cluster of minor genetic variants causing polygenic hypertriglyceridemia, which is exacerbated by conditions or medications which increase triglyceride levels beyond the saturation point of triglyceride removal systems. This situation is termed the multifactorial chylomicronemia syndrome. These aggravating factors include common conditions such as uncontrolled diabetes, overweight and obesity, alcohol excess, chronic kidney disease and pregnancy and several medications, including diuretics, non-selective beta blockers, estrogenic compounds, corticosteroids, protease inhibitors, immunosuppressives, antipsychotics, antidepressants, retinoids, L-asparaginase, and propofol. A third uncommon cause of the chylomicronemia syndrome is familial forms of partial lipodystrophy. Development of pancreatitis is the most feared complication of the chylomicronemia syndrome, but the risk of cardiovascular disease as well as non-alcoholic steatohepatitis is also increased. Treatment consists of dietary fat restriction and weight reduction combined with the use of triglyceride lowering medications such as fibrates, omega 3 fatty acids and niacin. Effective management of aggravating factors such as improving diabetes control, discontinuing alcohol and replacing or reducing the dose of medications that raise triglyceride levels is essential. Importantly, many if not most cases of the chylomicronemia syndrome can be prevented by effective identification of polygenic hypertriglyceridemia in people with conditions that increase its likelihood or before starting medications that may increase triglyceride levels. Several new pharmacotherapeutic agents are being tested that are likely to considerably improve treatment of hypertriglyceridemia in people at risk.

S1423 Lipoprotein Lipase Deficiency as a Rare Cause of Acute Pancreatitis

INTRODUCTION: Hypertriglyceridemia (HTG)-induced pancreatitis comprises 1 to 14 % of all cases of acute pancreatitis. Generally, primary HTG refers to a genetic disorder that includes type 1, 4 and 5 dyslipidemia. Familial lipoprotein lipase (LPL) deficiency is a rare condition of primary HTG with autosomal recessive transmission and a prevalence of 1-2:1,000,000. This case describes a patient with acute pancreatitis attributed secondary to LPL deficiency. CASE DESCRIPTION/METHODS: A 28 year old female with a history of type 1 diabetes mellitus complicated by diabetic gastroparesis s/p gastrojejunal bypass, HTN, HTG, recurrent pancreatitis presented to ED with abdominal pain. She had numerous admissions in the past due to recurrent pancreatitis secondary to HTG , one of which was complicated by necrotizing pancreatitis and wall off necrosis for which she underwent partial pancreatectomy. The patient also has an extensive family history of early onset type 1 diabetes and HTG. Despite the use of high intensity statin, fenofibrate, diet restriction and insulin pump, triglyceride levels has been very difficult to control. On physical examination, multiple red-yellowish bumps are noted on the arms consistent with xathomas. Labs were significant for elevated triglycerides and lipase (4500 and 400 respectively). Abdomen and pelvis CT scan showed evidence suggestive of acute pancreatitis, splenic vein thrombosis and hepatosplenomegaly. Due to high suspicion, molecular genetic testing for LPL deficiency was sent and came back positive. DISCUSSION: Patients with LPL are classically found to have high serum plasma triglycerides, low HDL-cholesterol, and presence of fasting chylomicrons in the plasma due to mutation of the enzyme that is responsible for hydrolysis and removal of triglycerides from the plasma. The most feared complication of the disease is recurrent and potentially life threatening acute pancreatitis. Although this is a very rare condition, it is an important differential diagnosis to consider due to refractoriness of conventional medical therapies including statins, fibrates and reduced fat intake(less than 20 g/day). There have been some studies that suggested high dose antioxidant therapy could potentially help lower the level of triglycerides and prevent recurrent episodes of acute pancreatitis in that particular group of patients. It is also critical to make the diagnosis as soon as possible for early medical interventions during childhood and genetic counseling and testing for family members.

EarLy Elimination of Fatty Acids iN hypertriglyceridemia-induced acuTe pancreatitis (ELEFANT trial): Protocol of an open-label, multicenter, adaptive randomized clinical trial

IntroductionAcute pancreatitis (AP) is a life-threatening inflammatory disease, with no specific pharmacological treatment. However, concerning some etiologies, early specific intervention (such as ERCP in biliary AP) has proven to be remarkably beneficial. Hypertriglyceridemia (HTG) induces severe pancreatic damage by several direct (cellular damage) and indirect (deterioration of microcirculation) mechanisms. Published data suggest that early removal of triglycerides (TGs) and toxic free fatty acids (FFAs) may be advantageous; however, high-quality evidence is still missing in the literature. MethodsDesign: ELEFANT is a randomized controlled, multicenter, international trial testing the concept that early elimination of TGs and FFAs from the blood is beneficial in HTG-AP. The study will be performed with the adaptive “drop-the-loser” design, which supports the possibility of dropping the inferior treatment arm, based on the results of the interim analysis. Patients with HTG-AP defined by TG level over 11.3 mmol/l (1000 mg/dL) are randomized into three groups: (A) patients who undergo plasmapheresis and receive aggressive fluid resuscitation; (B) patients who receive insulin and heparin treatment with aggressive fluid resuscitation; and (C) patients with aggressive fluid resuscitation. Please note that all intervention must be started within 48 h from the onset of abdominal pain. Exclusion criteria are designed logically to decrease the possibility of any distorting effects of other diseases. The composite primary endpoint will include both severity and mortality. ResultsOur null hypothesis is that early elimination of HTG and FFAs reduces the risk of mortality and severity of AP. Sample size calculation suggests that 495 patients will need to be enrolled in order to confirm or reject the hypothesis with a 10% dropout, 80% power and 95% significance level. The general safety and quality checks required for high-quality evidence will be adhered to. The study will be organized between February 2020 and December 2025. ConclusionOur study would provide the first direct evidence for or against early intervention in HTG-induced AP.

Fe3O4@OA@Poloxamer nanoparticles lower triglyceride in hepatocytes through liposuction effect and nano-enzyme effect

Lipid deposition induced various diseases including nonalcoholic fatty liver and hyperlipemia. The excessive accumulation of triglyceride (TG) and the deposition of fat were the two most critical causes. Here, we developed Fe3O4@OA@Poloxamer nanoparticles (NPs) with amphiphilic structures, which exhibited an excellent role in eliminating excess TG. Hydrophobic TG was adsorbed efficiently by Fe3O4@OA@Poloxamer NPs through the “liposuction effect” and the formation of NPs@TG complex was then conducted. The NPs@TG complex was further enclosed by the endosome based on the endocytosis and subsequently was taken into the lysosome, degrading with the help of lipases. Meanwhile, the “nano-enzyme effect” of Fe3O4 NPs recovered the lipid-regulated proteins including PPARα, further triggering biodegradation pathways of TG, although the lipid-regulated proteins were obviously inhibited in the high-fat hepatocytes models. These two mechanisms of Fe3O4@OA@Poloxamer NPs together achieved the down-regulation of TG in vivo and in vitro. Therefore, our findings provided a novel thought in treating these diseases associated with lipid deposition, that is, nanoparticles modified by specific structure exhibit a superior TG removal.

Efficient separation of triglyceride from blood serum using Cinnamon as a novel biosorbent: Adsorption thermodynamics, kinetics, isothermal and process optimization using response surface methodology

Abstract Batch adsorption of triglyceride (TG) from the human blood serum was investigated using cinnamon as a novel serum-contact biosorbent. Important factors affecting the adsorption including; temperature, contact time, stirring rate, adsorbent dose and, particle size were investigated through optimization using central composite design methodology. SEM and FTIR analyses were employed to characterize the adsorbent. The Langmuir, Freundlich, Temkin, and Dubinin-Radushkevitch isotherm models were employed to study the adsorption equilibria and the Langmuir and D-R models showed better agreement with the experimental data. The maximal TG adsorption and removal percentage values were determined as 18.12 mg/g and 75.16%, respectively. The pseudo-first order, pseudo-second order, intra-particle diffusion, and Elovich models were used to describe the adsorption kinetic and the pseudo-second order model better represented the data. Gibbs free energy and enthalpy changes were determined as -22.64 and 20.21 kJ/mol, respectively. Optimization results demonstrated that the optimal conditions for the adsorption were the adsorbent dose of 0.94 g, milling time of 9.19 min, stirring rate of 255 rpm, temperature of 23.41 °C and, contact time of 35 min which resulted in the optimal TG removal (88.2%). Based on the results, cinnamon is an efficient triglyceride reducer for possible application in extracorporeal blood perfusion.

Plasmapheresis for hypertriglyceridemia: The association between blood viscosity and triglyceride clearance rate.

Several factors in double filtration plasmapheresis (DFPP) were associated with triglyceride (TG) clearance rate. This study examines whether baseline whole blood viscosity was a predictor for efficient TG removal. Adult subjects who receiving DFPP for hyperlipidemia in Taoyuan General Hospital from January 2015 to March 2018 were classified into efficient and inefficient TG removal according to TG removal rate ≥50% vs <50%. TG removal rate was defined as following formula: (pre-apheresis TG- post-apheresis TG)/pre-apheresis TG. Whole blood viscosity (WBV) was estimated by following equation: WBV=0.12×hematocrit +0.17×(total protein -2.07). Univariate linear regression was used to assess the association between TG removal rate and WBV. Odds ratios (ORs) and 95% confidence interval (95%CI) for associations between variables and efficient TG removal were evaluated by logistic regression model to including univariate and multivariate adjustment. From a total of 66 subjects receiving DFPP, 37 subjects reached efficient TG removal. The difference in pre-apheresis TG levels, hematocrit, and WBV between efficient vs. inefficient TG removal groups was 4.1 vs 6.7mmol/L; 43.1% vs 39.5%; and 6.0cP vs 5.cP (Ps <0.05). After multivariate adjustment, WBC was a significant predictor for efficient TG removal (ORs and 95% CI were 3.192 (1.300-7.838), P<0.05). The correlation between WBV and extraction of TG was significant (r=-0.255, P=0.039). Hyperviscosity reduced the efficiency of TG removal in those receiving DFPP.

Effect of Extraction Time on Unreacted Oil Removal in Biodiesel Purification Using Deep Eutectic Solvent

Rice bran oil (RBO) based biodiesel contains unreacted oil such as monoglyceride (MG), diglyceride (DG) and triglyceride (TG) to be purified. The liquid-liquid extraction (LLE) method was used for purification using Deep Eutectic Solvent (DES). The objective of this work was to study the effect of extraction time on unreacted oil removal. RBO containing 16.49% oil with free fatty acids (FFA) content of 44.75%. Acid catalyzed methanolysis was used for biodiesel production under operating conditions: T = 60°C, t = 8 hours, molar ratio of oil/methanol was 1/10, H2SO4 1% w/w of RBO. Crude biodiesel containing 89.05% fatty acid methyl ester (FAME), 0.05% FFA, TG 4.03%, DG 4.01% and MG 0.30%. DES was made from choline chloride and ethylene glycol with 1/2 molar ratio, while molar ratio of biodiesel/DES was 1/2. The extraction time was varied from 15 to 240 minutes at 30°C. The highest TG, DG and MG removal were obtained at 240 minutes, they were 3.01%, 0.22% and 0.03%, respectively. FAME and FFA content were 96.55% and 0.03%. Keywords: biodiesel; DES; extraction; unreacted oil; purification

Development of a Fatty Liver Model by Restricted Feeding of Lactating Sheep

Background: As a frequent subclinical disease, fatty liver disease (FLD) is associated with a severe negative energy balance (NEB) during the early lactation period, and usually cause of economic loss to dairy farmers. Liver biopsy is the gold standard for the assessment of FLD. However, as an invasive procedure, liver biopsy has several limitations and such procedures are not readily available to dairy farmers. To further evaluate FLD in dairy cows, a FLD model of lactating sheep was developed by simulation of the state of negative energy balance (NEB).Materials, Methods &amp; Results: Fourteen pregnancy thin-tail ewes were divided into control group (CG, n = 4), non-lamb restrained feeding group (NRG, n = 4) and single birth restrained feeding group (SRG, n = 6). After lambing, NRG and SRG ewe were received a feed restrained diet for 16 days. Liver biopsies and blood was collected on days 1, 4, 7, 10, 13, and 16, and biochemical parameters were analyzed. With restricted feeding and lactation administration, ewes in SRG showed increased liver fat concentrations (LFC) from days 4 post-administration and severe LFC was detected at day 13. Compared with CG, SRG sheep showed significant lower concentration of serum glucose (Glu) from days 7-13 and higher non-esterified fatty acid (NEFA) from days 4-16, β-hydroxybutyric acid (BHBA) from days 4-16, triglyceride from days 4-16, low-density lipoprotein cholesterol from days 4-16, lactate dehydrogenase (LDH) from days 13-16, aspartate aminotransferase (AST) at days 16. While, ewes in NRG showed normal LFC levels, and high concentration of serum Glu and insulin from days 4-16 were detected than CG and SRG ewes. With restricted feeding, ewes in NRG and SRG showed significant low level of revised quantitative insulin sensitivity check index from days 4-16 and high level of liver total cholesterol (TC) at day 16. Liver pathological characteristics showed LFC of NEB sheep was first detected around the liver portal area.Discussion: In this study, a model of FLD in lactating thin-tail sheep was developed by restricted feeding. Serum glucose concentrations were sharply decreased in SRG sheep，that was due to the large energy requirements for lactation and low energy provided by a restricted diet. While non-lactating NRG sheep demonstrated lower fat mobilization, which was considered to contribute to the high concentrations of serum glucose, as compared to SRG sheep. Meanwhile, in a state of NEB, oxaloacetic acid, which is generated by glycolysis and glycogenic amino acids, tends to be used for gluconeogenesis, that a generous amount of NEFA is incompletely oxidized to generate ketone body in SRG sheep, which is a major component of BHBA. Liver TC concentrations were significantly higher in NRG sheep than those in the SRG sheep, while liver triglyceride was significantly lower. The high level of liver TC in NRG sheep was considered to induce removal of triglyceride from the liver in the form of VLDL. Compared with CG sheep, although higher levels of liver TC were detected in SRG sheep on postpartum day 16, these levels were considered too low to induce significant depletion of triglycerides from the liver. In this study, the increase in serum AST and LDH was considered to cause by oxidative stress in mitochondria, and LDH concentrations was considered more sensitively than AST for LFC caused by NEB. Liver pathological characteristics showed that FLD caused by NEB had a major impact on reduced LFC, although no significant liver fibrosis was detected. While different from FLD caused by high-fat diet, TG was first accumulates around the hepatic lobules and LFC of NEB sheep was first detected around the liver portal area. It was considered that high concentrations of NEFA are prioritized for oxygenation in the liver portal area, which results in triglyceride accumulation.

The culture or co-culture of Candida rugosa and Yarrowia lipolytica strain rM-4A, or incubation with their crude extracellular lipase and laccase preparations, for the biodegradation of palm oil mill wastewater

Wastewater generated by the palm oil industry contains a high concentration of oil, grease and phenolic compounds, which makes it difficult to treat. The aim of this research was to evaluate the ability of free Candida rugosa and Yarrowia lipolytica rM-4A yeast cell cultures, as a single yeast strain and as a co-culture, as well as their crude extracellular enzyme preparations, to reduce the total phenolic compounds, triglycerides and chemical oxygen demand (COD) level in palm oil mill effluent (POME) that contains high amounts total phenolic compounds (9783 mg L−1) and oil and grease (7762 mg L−1). Culture with Y. lipolytica rM-4A effectively reduced the total phenolic content in undiluted POME, resulting in removal of 36% of the total phenolic content after 96 h. The highest removal of triglyceride (98.5%) and COD (60.3%) were observed in undiluted POME treated by the co-culture of C. rugosa and Y. lipolytica for 120 h. Triglyceride was removed rapidly by the extracellular lipase produced by C. rugosa, reaching 93% degradation after 48 h. This research revealed the excellent potential of both crude extracellular enzyme preparations and the co-culture of the two yeasts in the pretreatment of high fat and oil containing POME.

Butter Blood

Acute pancreatitis induced by hypertriglyceridemia is a rare but well-established complication, though the exact pathogenesis remains unclear. Estimated rates account for 1-4% of acute pancreatitis cases with noticeable rising risk as serum levels increase (triglycerides > 1000 carry 5% risk; triglycerides > 2000 carry 10-20% risk). Occurrence is more commonly associated in those with acquired disorders (such as diabetes mellitus, hypothyroidism, obesity, pregnancy, medication-induced, nephrotic syndrome) than in those with inborn errors of lipoprotein metabolism (such as familial combined hyperlipidemia, familial hypertriglyceridemia). Therapeutic measures in such patients include standard pancreatitis management, along with lipidlowering oral agents, low-molecular weight heparin, and insulin in those who are diabetic. In situations where these measures fail to improve patient clinic course, plasma exchange is used to expedite removal of triglycerides (TG). Here, we present a case of a noncompliant, insulin-dependent diabetic patient found to have hypertriglyceridemia-induced pancreatitis. Case: A 40-year-old African American male with poorly-controlled, insulin-dependent diabetes presented to our facility with complaints of abdominal pain accompanied with nausea and vomiting. Laboratory examination was consistent with severe pancreatitis and mild ketoacidosis (Table 1). Abdominal CT scan confirmed the clinical diagnosis of severe pancreatitis of the pancreatic head and uncinate process with inflammation of the adjacent duodenum (Image 1-2). The appearance of latescent serum accompanied with lipemic serum pointed toward a diagnosis of pancreatitis secondary to hypertriglyceridemia.Figure 1Figure 2Figure 3As our patient did not carry a familial history of genetic abnormalities of lipoprotein metabolism, we attributed the hypertriglyceridemia secondary to his poorly-controlled diabetes and diabetic ketoacidosis. The patient was treated with gemfibrozil, atorvastatin, IV fluids, bowel rest, and insulin infusion. Improvement of his triglyceride level coincided with improvement in blood sugars and abdominal pain. Plasmapharesis was not undertaken in this patient given improvement with conventional therapies.

Abstract 537: Characterization of the Lipase Stimulating Domain for Apolipoprotein A-V and the Development of a Therapeutic Peptide for the Treatment of Hypertriglyceridemia

The impaired clearance of plasma triglycerides is a major cause of clinical hypertriglyceridemia, a condition which affects about one third of adults in the G7 countries and is a risk factor for a number of diseases, including cardiovascular disease, cancer and pancreatitis. The hydrolysis and removal of triglycerides from the plasma is dependent on cell surface lipoprotein lipase (LpL) and its enzymatic activity is in turn regulated by specific apolipoproteins; apoC-III, which acts as a potent inhibitor and apoC-II and apoA-V, which act as stimulators. Their importance in hypertriglyceridemia is underscored by GWAS analyses, which have identified the APOA1/C3/A4/A5 gene cluster (chromosome 11q23) as an important determinant of dylipidemia. ApoA-V is of particular interest since its physiological concentration (3.8 nM) is 10,000x lower than that of apoC-II and the other major apolipoproteins. To investigate the influence of different lipoproteins on lipase kinetics we have developed a fluorescence-based assay that uses a fluorogenic triglyceride analog (EnzChek®) in a microplate format. Here we report that a short alpha-helical amphipathic peptide (AV199-232) corresponding to apoA-V residues 199-232 exhibit; i) lipid-binding, ii) heparin-binding and iii) lipase-stimulating activities. Structural characterization and residue substitution studies have been employed to map these three activities to three separate discrete domains within AV199-232. Enzyme kinetic analysis with pure LpL indicates that AV199-232 can overcome the inhibition of LpL by apoC-III and stimulate enzyme activity by reducing its Km 8-fold. Similar increases in lipase activity are also observed with live THP-1 monocytes and HepG2 cells that express hepatic lipase. The influence of AV199-232 in cell culture is saturable, dependent on the presence of cell surface lipase, and enzymatic activation can be sustained for at least 2 hrs. We postulate that apoA-V and AV199-232 stimulate lipase activity through an allosteric mechanism by binding to lipases with high affinity near their catalytic site to affect enzyme kinetics.