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The subendothelial retention of apolipoprotein B (apoB)-containing lipoproteins is a critical step in the initiation of pro-atherosclerotic processes. Recent genetic and clinical evidence strongly supports the concept that the lipid content of the particles is secondary to the number of circulating atherogenic particles that are trapped within the arterial lumen. Since each low-density lipoproteins (LDL) particle contains one apoB molecule, as do intermediate density lipoprotein (IDL) and very low-density lipoprotein (VLDL) particles, apoB level represents the total number of atherogenic lipoproteins, which is independent of particle density, and not affected by the heterogeneity of particle cholesterol content (clinically evaluated by LDL-cholesterol level). From this perspective, apoB is proposed as a better proxy to LDL-cholesterol for assessing atherosclerotic cardiovascular disease risk, especially in specific subgroups of patients, including subjects with diabetes mellitus, with multiple cardiometabolic risk factors (obesity, metabolic syndrome, insulin resistance, and hypertension) and with high triglyceride levels and very low LDL-cholesterol levels. Therefore, given the causal role of LDL-cholesterol in atherosclerotic cardiovascular disease (ASCVD) development, routine measurement of both LDL-cholesterol and apoB is of utmost importance to properly estimate global cardiovascular risk and to determine the 'residual' risk of ASCVD in patients receiving therapy, as well as to monitor therapeutic effectiveness.

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Human female infertility, 20% of which is idiopathic, is a public health problem for which better diagnostics and therapeutics are needed. A novel cause of infertility emerged from studies of female mice deficient in the HDL receptor gene (Scarb1). These mice are infertile and have high plasma HDL cholesterol (C) concentrations, due to elevated HDL-free cholesterol (FC), which transfers from HDL to all tissues. Previous studies have indicated that oral delivery of probucol, an HDL-lowering drug, to female Scarb1-/- mice reduces plasma HDL-C concentrations and rescues fertility. Additionally, serum opacity factor (SOF), a bacterial virulence factor, disrupts HDL structure, and bolus SOF injection into mice reduces plasma HDL-C concentrations. Here, we discovered that delivering SOF to female Scarb1-/- mice with an adeno-associated virus (AAVSOF) induces constitutive SOF expression, reduces HDL-FC concentrations, and rescues fertility while normalizing ovary morphology. Although AAVSOF did not alter ovary-FC content, the ovary-mol% FC correlated with plasma HDL-mol% FC in a fertility-dependent way. Therefore, reversing the abnormal plasma microenvironment of high plasma HDL-mol% FC in female Scarb1-/- mice rescues fertility. These data provide the rationale to search for similar mechanistic links between HDL-mol% FC and infertility and the rescue of fertility in women by reducing plasma HDL-mol% FC.

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Immune and inflammatory mediators in autoimmune rheumatic diseases induce modification in the activity of enzymes pivotal for lipid metabolism and promote a proatherogenic serum lipid profile. However, disturbances in low- and high-density lipoprotein composition and increased lipid oxidation also occur. Therefore, lipoprotein dysfunction causes intracellular cholesterol accumulation in macrophages, smooth muscle cells, and platelets. Overall, both plaque progression and acute cardiovascular events are promoted. Single rheumatic diseases may present a particular pattern of lipid disturbances so that standard methods to evaluate cardiovascular risk may not be accurate enough. In general, antirheumatic drugs positively affect lipid metabolism in these patients.

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To increase our understanding of factors contributing to therapeutic failure (TF) in leishmaniasis, we have studied some plasma membrane features of host THP-1 cells infected with clinical isolates of Leishmania infantum from patients with leishmaniasis and TF. The fluorescent probes DPH and TMA-DPH were used to measure changes in membrane fluidity at various depths of the plasma membranes. Steady-state fluorescence anisotropy of DPH embedded in the infected THP-1 membranes showed a significant increase, thereby suggesting a substantial decrease in plasma membrane fluidity relative to controls. Considering that cholesterol affects membrane fluidity and permeability, we determined the cholesterol content in plasma membrane fractions of human macrophages infected with these L. infantum lines and observed a significant increase in cholesterol content that correlates with the measured decrease in plasma membrane fluidity. In order to define the pathways that could explain the increase in cholesterol content, we studied the transcriptomics of the cholesterol-enriched pathways in host THP-1 cells infected with TF clinical isolates by RNA-seq. Specifically, we focused on four enriched Gene Ontology (GO) terms namely cholesterol efflux, cholesterol transport, cholesterol metabolic process and cholesterol storage. Additionally, we analyzed the genes involved in these pathways. Overall, this study shows that these clinical isolates are able to modulate the expression of specific genes in host cells, thereby modifying the cholesterol content in plasma membranes and inducing changes in plasma membrane fluidity that could be associated with the parasite's ability to survive in the host macrophages, thereby possibly contributing to immune evasion and TF.

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The biofortification of basal laying hen feed with natural matrices can improve the beneficial potential of eggs produced without relying on artificial fortification. The present study aimed to evaluate the effects of hen diet supplementation with dried Moringa leaves (DML) and goji berries (DGB) on egg functional properties in terms of cholesterol and carotenoid content. Forty Lohman Brown Classic laying hens were randomly divided into four groups. The control group (G1) received the basal poultry diet, group G2 received a diet with 5% DML + 10% DGB, group G3 received a diet with 3% DML + 7% DGB, and group G4 received a diet with 15% DML. HPLC-DAD analysis showed that feed supplementation positively influenced the egg carotenoid content, with a valuable increase in xanthophylls concentration, especially lutein (+333.24% in G4, +258.15% in G2, +189.24% in G3, compared to G1). The same trend was also followed by the ß-carotene concentration (+181.38% in G3 and +116.01% in G4, compared to G1). Furthermore, the eggs obtained from G3 showed the lowest cholesterol content (-47.08%). Additionally, the performed antioxidant assays showed maximum activity in G2 (+39.11 compared to G1 for the DPPH test) and in G4 (+31.11 compared to G1 for the ABTS test). In conclusion, the G2 experimental diet could be potentially used in poultry industries to produce "functional eggs".

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The present study was designed to study the effects of conventional and free range systems on egg quality attributes, fatty acid profile, and cholesterol contents of laying hen. Two hundred and forty R1 cross of Rhode Island Red (RIR) × Fyoumi (F) layers during the peak production were divided into two groups and reared in two different housing systems (free range vs. conventional systems). The statistical analysis of the results revealed that the housing system (modification of diet) significantly (p < 0.05) affected egg quality, fatty acid profile, and cholesterol contents. The highest (p < 0.05) whole egg and albumen weights were recorded for eggs produced in a conventional system and higher yolk weight and shell thickness were recorded in free range hens. Significantly (p < 0.05) lower concentration of saturated and higher concentration of mono- and polyunsaturated fatty acids were observed in eggs produced under a free range system. In addition, higher level of total omega-3 fatty acids and higher ratio of omega-6 to omega-3 fatty acids were observed in free range eggs as compared to those in the conventional system. From the results of the present study, we concluded that eggs produced from Rhode Island Red × Fyoumi laying hens in a free range system are qualitatively superior compared to those in the conventional system.

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Encapsulation in liposomes has been an efficient strategy to improve the stability of sensitive bioactive compounds such as essential oils (EOs). However, the stability of liposomal formulations remains a key parameter controlling the delivery of encapsulated ingredients. Cholesterol (Chol) modulates the membrane properties conferring stability to the lipid bilayer. Thus, the Chol content in the liposome formulations encapsulating EO components should be carefully chosen. In this work, various liposome formulations differing by Chol content (DPPC:Chol 100:10; 100:25; 100:50; 100:75; 100:100) were exposed to a series of 22 EO components at DPPC/EO 100/25. The formulations were characterized for their final composition and their permeability to the hydrophilic fluorophore, sulforhodamine B (SRB), was monitored. Results showed that the Chol content experimentally determined for the various formulations (above 10% Chol) was below the theoretical weighed Chol. Among the tested components, 13 molecules displayed a significant permeabilizing effect on 10% Chol membranes. Most of these possess a hydroxyl group. The EO induced permeability was dependent on the Chol content which affects the membrane phase: their effect was reduced upon increasing Chol content keeping five EOs components effective at 40% Chol. The EO's effect was also linked to the hydrophobicity of the molecule. Hence, the DPPC:Chol ratio of the formulation is chosen considering the structure of the compound, its hydrophobicity and its effect on the permeability at different Chol content: a formulation comprising 40% Chol is suggested for highly hydrophobic molecules whereas a formulation with higher Chol content could be selected for less hydrophobic compounds.

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Coenzyme Q10 (CoQ, ubiquinone) is a redox-active lipid endogenously synthesized by the cells. The final stage of CoQ biosynthesis is performed at the mitochondrial level by the 'complex Q', where coq2 is responsible for the prenylation of the benzoquinone ring of the molecule. We report that the competitive coq2 inhibitor 4-nitrobenzoate (4-NB) decreased the cellular CoQ content and caused severe impairment of mitochondrial function in the T67 human glioma cell line. In parallel with the reduction in CoQ biosynthesis, the cholesterol level increased, leading to significant perturbation of the plasma membrane physicochemical properties. We show that 4-NB treatment did not significantly affect the cell viability, because of an adaptive metabolic rewiring toward glycolysis. Hypoxia-inducible factor 1α (HIF-1α) stabilization was detected in 4-NB-treated cells, possibly due to the contribution of both reduction in intracellular oxygen tension and ROS overproduction. Exogenous CoQ supplementation partially recovered cholesterol content, HIF-1α degradation, and ROS production, whereas only weakly improved the bioenergetic impairment induced by the CoQ depletion. Our data provide new insights on the effect of CoQ depletion and contribute to shed light on the pathogenic mechanisms of ubiquinone deficiency syndrome.

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The effects of organic Zn-L-selenomethionine (Zn-L-SeMet) at 0.3 ppm on carcass composition, meat characteristics, fatty acid composition, glutathione peroxidase activity, and ribonucleotide content were compared against the commercial inorganic sodium selenite (Na-Se) and the combination of the two, in commercial broilers. A total of 540 one day-old chicks were assigned at random to 3 dietary treatments : i) commercial inorganic selenium as control or T1, ii) a 1:1 ratio of inorganic and organic selenium as T2, and iii) organic selenium as T3. Carcass composition, meat characteristics, cholesterol content, fatty acid composition, and ribonucleotide content were generally unaffected by treatments. However, discrepancy were significantly observed in glutathione peroxidase activity (GSH-Px) and water holding capacity, with organic selenium showing higher glutathione peroxidase activity (p<0.01) and lower shrinkage loss (p<0.05), respectively. These findings could be explained by the contribution of organic selenium in bioavailability of GSH-Px. However, having conducted in a commercial close house system with sufficient amount of nutritional supplementation, the present study demonstrated little or no effects of organic Zn-L-SeMet on meat characteristics, fatty acid composition, and ribonucleotide content (flavor characteristic) in broiler chickens.

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The study was conducted to evaluate breast meat quality attributes of Marshal and FUNAAB Indigenous (Alpha C13 and Alpha C14) Broiler (FIB) chickens. Birds of average live weights (1.8 to 2.0 kg) were selected from flock fed same concentrate diet and managed under deep litter system. Birds were slaughtered, scalded in hot water at 65 °C, de-feathered, and frozen-stored. Proximate, mineral, cholesterol contents, pH, color, and fatty acid profile of the chickens' meat were determined. Data were subjected to ANOVA (p < 0.05). Moisture (73.87%) and fat (2.82%) contents of Marshal were higher than FIB. Protein (23.65%) and ash (1.56%) were highest (p < 0.05) for Alpha C14. pH ranged between 5.94 and 6.58. Lightness (L*) and redness (a*) values were 69.37-75.52 and 8.77-7.96. Alpha C13 had significantly higher L* and a* than Marshal. Potassium was most abundant mineral observed across breeds. Saturated (SFA, 38.03-57.10%), monounsaturated (MUFA, 29.78-41.55%), and polyunsaturated (PUFA, 10.91-20.29%) fatty acid compositions differed significantly among breeds. Cholesterol contents of Alpha C14 (4.23 mg/kg) was significantly lower, while Marshal (5.67 mg/kg) chicken showed significantly higher value. Conclusively, FIB chickens are valuable nutritional meat sources with desirable chemical composition and can constitute alternative chicken meats with human health benefit.

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In cholesterol/DSPE-PEG2000/DPPC systems, nano-disks could evolve between liposomes and spherical micelles in a certain range of PEG-lipids. How cholesterol or drug influences this evolution and what about the properties of discoidal micelles as drug carrier are still not clear. Aiming at this, nanocarriers with different contents of cholesterol and DSPE-PEG2000 were prepared by thin-film-hydration method. Firstly, the bilayer fluidity of nanocarriers was investigated and proved to decrease with the increase of cholesterol, and the cooperative degree between phospholipids was also related to cholesterol content in an order of 30 > 40 > 15 > 0 mol%. Then three different levels of cholesterol were chosen to study its effect on DSPE-PEG2000 limit used to form discoidal micelles. For transition from liposomes to disks, the limit of DSPE-PEG2000 employed is above 0.5 mol% at 0 mol% cholesterol, above 15 mol% at 30 mol% cholesterol and above 5 mol% at 40 mol% cholesterol observed from TEM images, indicating that nanocarriers with 30 mol% cholesterol formed discoidal micelles most difficultly due to the strongest interaction between phospholipids. However, membrane fluidity seems to have little responsibility for the different morphologies. And imatinib (IM) could promote the formation of discoidal micelles, resulting from the interaction between IM and polar headgroups of phospholipid demonstrated by DSC. The relative reduction of entrapment efficiency for three agents with different dissolving properties showed that lipophilic drugs were the most suitable drug that could be loaded into discoidal micelles. In addition, the discoidal micelle formulations tested could readily change to other morphologies after 80 days' storage. In conclusion, both the cholesterol content and IM could affect the formation of discoidal micelles by probably influencing the interaction between phospholipids, also the discoidal structure was not stable enough and only suitable for lipophilic drug loading. We hope this study could help to design the formula and choose the proper drugs that may retain the discoidal morphology.

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To transfer the viral genome into the host cell cytoplasm, internalized influenza A virus (IAV) particles depend on the fusion of the IAV envelope with host endosomal membranes. The antiviral host interferon (IFN) response includes the upregulation of interferon-induced transmembrane protein 3 (IFITM3), which inhibits the release of the viral content into the cytosol. Although IFITM3 induction occurs concomitantly with late endosomal/lysosomal (LE/L) cholesterol accumulation, the functional significance of this process is not well understood. Here we report that LE/L cholesterol accumulation itself plays a pivotal role in the early antiviral defense. We demonstrate that inducing LE/L cholesterol accumulation is antiviral in non-IFN-primed cells, restricting incoming IAV particles and impairing mixing of IAV/endosomal membrane lipids. Our results establish a protective function of LE/L cholesterol accumulation and suggest endosomal cholesterol balance as a possible antiviral target.IMPORTANCE With annual epidemics occurring in all parts of the world and the risk of global outbreaks, influenza A virus (IAV) infections remain a major threat to public health. Infected host cells detect viral components and mount an interferon (IFN)-mediated response to restrict virus propagation and spread of infection. Identification of cellular factors and underlying mechanisms that establish such an antiviral state can provide novel strategies for the development of antiviral drugs. The contribution of LE/L cholesterol levels, especially in the context of the IFN-induced antiviral response, has remained controversial so far. Here, we report that accumulation of cholesterol in the LE/L compartment contributes to the IFN-induced host cell defense against incoming IAV. Our results establish cholesterol accumulation in LE/L per se as a novel antiviral barrier and suggest the endosomal cholesterol balance as a putative druggable host cell factor in IAV infection.

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Esterified cholesterol content is often lower than free cholesterol content in biological systems and thus the determination of the esterified cholesterol content of cells is often challenging. Traditional methods use enzymatic assays in which an indirect measurement of the esterified cholesterol content is obtained by subtracting the measurements of the free from the total cholesterol content. However, this approach fails in the case where the total cholesterol content of cells is unchanged while the ratio of free to esterified cholesterol changes such that total and free cholesterol content are very similar and thus the difference may fall within the background noise of the enzymatic assay. To overcome this challenge, we here describe a method that utilizes a TLC-based technique to isolate esterified cholesterol. Isolated esterified cholesterol can then be measured using traditional enzymatic methods. Therefore, this method provides a practical and more sensitive assay to measure esterified cholesterol content in cellular extracts.

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This study evaluates the effects of betaine supplementation (1gkg-1 for 20weeks) on the regulation of genes involved in lipid and cholesterol metabolism of Longissimus lumborum and Biceps femoris from obese Alentejano pigs. Betaine supplementation led to an increase in total cholesterol in both muscles, complementing results previously published indicating a significant increase on the intramuscular lipid content. The expression of twelve genes involved in lipogenesis, lipolysis/FA oxidation, FA transport, and cholesterol metabolism, as well as two transcription factors were also evaluated. Genes related to lipid and cholesterol synthesis plus FA transport were consistently up-regulated in both muscles of betaine fed pigs. On the other hand, genes related to lipolysis/FA oxidation were not affected or down-regulated by betaine supplementation. Our data suggest that the underlying mechanism regulating IMF and cholesterol accumulation in Alentejano pigs supplemented with betaine is associated with the up-regulation of genes involved in lipid synthesis, FA transport, and cholesterol synthesis.

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A study was conducted using 144 laying hens to evaluate the effects of adding aqueous extract of Tribulus terrestris to diets on productive performance, egg quality traits, and some blood parameters of laying hens reared under cold stress condition (6.8 ± 3 °C). The birds were randomly assigned to each of four dietary treatments (C, T1, T2, and T3) with six replicate cages of six birds. Diet inclusion of aqueous extract of T. terrestris at the rate of 10, 20, and 30 ml/Lit offered to groups T1, T2, and T3, respectively, while group C served as the control diet with no addition. Feed intake (FI), feed conversion ratio (FCR), egg weight (EW), egg production (EP), and egg mass (EM) were evaluated during the 42-day trial period. The EP and EM increased, whereas FCR decreased (P < 0.001) in the hens fed the extract-included diet as compared to those fed the basal diet. The serum content of cholesterol decreased and the thickness of egg shell increased in the hens fed the T2 and T3 diet compared to those fed the basal diet. Overall from the results of the present experiment, it can be concluded that diet supplementation with aqueous extract of T. terrestris has beneficial effects on productive performance of laying hens reared under cold stress condition.

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O objetivo deste experimento foi avaliar a influência da variação do nível de energia metabolizável na dieta de frangos de corte, associada a uma variação proporcional da densidade nutricional da ração, sobre o desempenho das aves e sobre a composição da fração lipídica da carne. Foram avaliados separadamente machos e fêmeas da linhagem Cobb 500. Para análise de desempenho, foi utilizado um delineamento inteiramente casualizado em esquema fatorial 6x3, sendo seis níveis de energia metabolizável na ração (2.800, 2.900, 3.000, 3.100, 3.200 e 3.300kcal kg-1) e três idades ao abate (42, 49 e 56 dias). Foi utilizada a metodologia de superfície de resposta para estabelecer um modelo matemático, explicando o comportamento do peso vivo, do consumo de ração e da conversão alimentar. Para a avaliação da composição da fração lipídica da carne, determinou-se o teor de lipídeos totais e de colesterol na carne do peito, na coxa com pele e na coxa sem pele, seguindo um delineamento inteiramente casualizado em esquema fatorial 3x3x2, sendo três níveis de energia metabolizável na ração (2.800, 3.000 e 3.300kcal kg-1 de ração), três idades ao abate (42, 49 e 56 dias) e dois sexos. A redução da energia metabolizável da ração até valores próximos de 3.000kcal kg-1 não afetou o peso vivo, mas, reduzindo a energia da ração abaixo desse valor, o peso vivo diminui. O consumo de ração foi menor quando o nível energético da ração foi mais alto. A conversão alimentar melhorou de forma proporcional ao aumento do nível energético da ração. O desempenho de todas as aves ficou dentro da faixa considerada adequada para a linhagem. A carne de peito apresentou ...

The purpose of this experiment was to investigate the influence of changes in dietary metabolizable energy level, associated with a proportional nutrient density variation, on broiler chickens performance and on the lipid composition of meat. Males and females Cobb 500 broilers were evaluated separately. Performance evaluation followed a completely randomized design with factorial 6x3 arrangement - six energy levels (2,800, 2,900, 3,000, 3,100, 3,200 and 3,300kcal kg-1) and three slaughter ages (42, 49 and 56 days). Response surface methodology was used to establish a mathematical model to explain live weight, feed intake and feed conversion behavior. Total lipids and cholesterol were determined in breast meat and in thigh meat, with and without skin. For lipid composition analysis, a 3x3x2 factorial arrangement in a completely randomized design - three ration's metabolizable energy levels (2,800, 3,000 and 3,300kcal kg-1), three slaughter ages (42, 49 and 56 days) and two sexes - was used. The reduction in the diet metabolizable energy close to 3,000kcal kg-1 did not affect live weight, but below this value, live weight decreased. Feed intake was lower when the dietary energy level was higher. Feed conversion was favored in a direct proportion to the increase of the energy level of the diet. The performance of all birds was within the range considered appropriate for the lineage. Breast meat had less total lipids and cholesterol than thigh meat. Thigh with skin had more than the double of the amount of total lipids than skinless thigh, but the cholesterol content did not differ with the removal of the skin. Intramuscular fat content was lower in the meat from birds fed with lower energy level ration. This information may help to define the most appropriate nutritional management. Despite the decrease in bird's productive performance, the restriction of energy in broiler chickens feed may be a viable alternative, if the consumers are ...

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The effects of beef fat substitution with kashar cheese were studied in traditional Turkish fermented sausage; sucuk. Six sucuk formulations were prepared by replacing 0, 10, 20, 30, 40 and 50% of beef fat was substituted with kashar cheese. The fat substitution of fat with kashar cheese decreased fat content and increased protein content of the product that affected the chemical, physical and sensorial characteristics of products. Saturated fatty acid content increased and unsaturated, mono-unsaturated and poly-unsaturated fatty acids amount were decreased as the cheese amount increased. The formulation with 10% substitution of beef fat with cheese took the best sensory overall acceptability scores followed by 20% and control groups.