Increase In Cholesterol Content Research Articles

Unveiling the Potential of Silymarin, Spirulina platensis, and Chlorella vulgaris towards Cardiotoxicity via Modulating Antioxidant Activity, Inflammation, and Apoptosis in Rats

This study assessed the possible pharmacological effects of Chlorella vulgaris (Cg), Spirulina platensis (St), and silymarin (Sl) against thioacetamide (TA)-induced cardiotoxicity in rats, with a focus on their antioxidant, cardioprotective, and anti-inflammatory properties. The following is the random grouping of sixty male rats into six groups of ten animals each: the control (negative control), TA-intoxicated group (positive control; 300 mg/kg body weight (BW)), Sl + TA group (100 mg Sl/kg BW + TA), St + TA group (400 mg St/kg BW + TA), Cg + TA (400 mg Cg/kg BW + TA), and St + Cg + TA group (400 St + 400 Cg mg/kg BW + TA) were all administered for 30 days. At the start of the study, groups 2 through 6 were administered TA intraperitoneally at a dosage of 300 mg/kg BW for two consecutive days, with a 24 h gap between each dose, to induce cardiac damage. Blood samples were obtained to measure hematological parameters and perform biochemical assays, including lipid profiles and cardiac enzymes. For histopathology and immunohistochemistry determination, tissue samples were acquired. The current findings showed that TA injection caused hematological alterations and cardiac injury, as evidenced by greater serum levels of troponin I, creatine kinase-MB, and total creatine kinase (p < 0.05), as well as significantly elevated serum malondialdehyde and decreased serum total antioxidant capacity (p < 0.05) concentrations. Moreover, an increase in blood low-density lipoprotein and total cholesterol concentration (p < 0.05) was recorded in the TA group. There were alterations in the heart tissue’s histological structure of the TA group compared to the control ones. These alterations were characterized by vacuolar degeneration of myocytes, loss of cross striation, coagulative necrosis, and fibrosis of interstitial tissue, which was ameliorated by the supplementation of SI, St, and Cg. The TA-intoxicated group showed weak expression of B-cell lymphoma protein 2 (p < 0.05) and strong immunoreactivity of tumor necrosis factor-α and B-cell lymphoma protein 2-associated X (p < 0.05). However, the groups receiving Sl, St, and Cg experienced the opposite. The administration of Sl, St, Cg, and St + Cg along with TA significantly improved and restored (p < 0.05) erythrogram indices, including RBCs, hemoglobin, total leukocytic count, lymphocytes, and monocyte, to the normal control values. The administration of Sl, St, and Cg alleviated the cardiotoxicity caused by TA via reducing oxidative stress, inflammatory markers, and apoptosis in heart tissue. In summary, the current findings suggest that the treatment with Sl, St, and Cg was beneficial in ameliorating and reducing the cardiotoxicity induced by TA in rats.

Digestive utilization of Sahelian forages associated with cowpea haulms (Vingna unguiculata) and blood profile in Arabic rams

Digestive utilization of sahelian forages associated with Vingna unguiculata and blood profile study was conducted in scientific garden of the University of Sciences and Technology, Ati (JSUSTA). Three forages were used to assess in vivo digestibity in eighteen (18) rams weighing around 31.83±3.02 kg and displayed individually in metabolic cages in six (6) groups of three (3) animals. Control groups received 1000g of feed made up of Brachiaria deflexa, Dactyloctenium aegyptium and Echinocloa colona. While, experimental groups received basal feeds associated with 30% of Vingna unguiculata. The experiment was preceded by 14 days adaptation and 7 days of data collection during which, hundred (100g) grams of feeds and feces as well as urine (10 ml) were collected for chemical analysis and digestibility assessment. At the end of experimentation, two (2) ml of blood was collected from each animal before and after feeding for blood profile analysis. The results showed that the association of 30% Vingna unguiculata have significantly (p<0.05) increased intake of D. aegyptium and B. deflexa in Arab rams. In vivo digestibility of E. colona was significantly (p<0.05) higher in rams when associated or no with 30% of Vingna unguiculata. Blood profile analysis showed that E. colona associated with 30% of Vingna unguiculata induced significant (p<0.05) increase in blood protein, globulin and cholesterol concentration in Arabic rams.

Cascara kombucha potential as a functional food for fasting glucose regulation, cholesterol control, and liver function modulation: in vitro and in vivo study

SummaryUnhealthy dietary patterns are a major contributor to obesity worldwide, leading to various adverse metabolic effects such as elevated liver enzyme levels and increased cholesterol concentrations. This study investigates the potential of cascara kombucha as a functional beverage to provide health benefits in both in vitro and in vivo models. Our results indicate that kombucha fermentation enhances the bioactive compounds in cascara, including polyphenols and flavonoids. The in vitro study found that cascara kombucha effectively decreased free radicals (DPPH and ABTS) levels, α‐amylase, and lipase activity, suggesting its potential to act as an antioxidant and modulate carbohydrate and lipid metabolism. In a mouse model, oral administration of cascara kombucha at a dose of 300 mg/kg body weight for 28 days successfully decreased fasting blood glucose levels, improved liver health by reducing aspartate transaminase and alanine transaminase levels, and lowered serum cholesterol content induced by a fat‐enriched diet compared to the untreated group. The levels of superoxide dismutase and catalase, suppressed by the fat‐enriched diet, were restored to normal. Additionally, cascara kombucha downregulated genes associated with inflammation induced by a fat‐enriched diet, including tnf‐α, IL‐6, IL‐1β, and cox‐2. It also positively affected the balance of intestinal microflora in mice by reducing coliform density and enhancing probiotic populations. These findings highlight the potential of cascara kombucha as a promising approach to addressing health issues caused by poor dietary habits and metabolic imbalances.

Glial cholesterol redistribution in hypoxic injury in vitro influences oligodendrocyte maturation and myelination

Hypoxic insult to the fetal brain causes loss of vulnerable premyelinating oligodendrocytes and arrested oligodendrocyte differentiation. Astrocytes influence oligodendrocyte differentiation and the astrocytic response to hypoxia could affect oligodendrocyte maturation under hypoxia. To identify pathways by which astrocytes influence oligodendroglial maturation in hypoxic injury, human fetal neural stem cell-derived astrocytes were exposed to 0.2 % oxygen for 48 hours. Transcriptomic analysis revealed the upregulation of the cholesterol-biosynthesis pathway in hypoxia-exposed astrocytes. Hypoxia-exposed primary astrocytes and astrocytic cell line (SVG) showed increased expression of hydroxy-methyl-glutaryl-CoA reductase (HMGCR), squalene epoxidase (SQLE), apolipoprotein E (apoE) and ATP-binding cassette transporter 1 (ABCA1) on qPCR and Western blot. Hypoxic SVG also showed increased cholesterol content in cells and culture supernatants and increased cell surface expression of ABCA1. Interestingly hypoxia-exposed premyelinating oligodendrocytes (Mo3.13) showed reduced cholesterol along with decreased expression of HMGCR and SQLE on qPCR and Western blot. Exogenous cholesterol increased the differentiation of Mo3.13 as measured by increased expression of myelin basic protein (MBP) on flow cytometry. Hypoxia exposure resulted in increased cholesterol transport from astrocytes to oligodendrocytes in cocultures with BODIPY-cholesterol labelled SVG and membrane-labelled Mo3.13. As exogenous cholesterol enhanced oligodendrocyte differentiation, our findings indicate that increased cholesterol synthesis by astrocytes and transport to oligodendrocytes could supplement oligodendroglial maturation in conditions of hypoxic brain injury in neonates.

Lipid loss and compositional change during preparation of simple two-component liposomes

Liposomes are used as model membranes in many scientific fields. Various methods exist to prepare liposomes, but common procedures include thin-film hydration followed by extrusion, freeze-thaw, and/or sonication. These procedures can produce liposomes at specific concentrations and lipid compositions, and researchers often assume that the concentration and composition of their liposomes are similar or identical to what would be expected if no lipid loss occurred. However, lipid loss and concomitant biasing of lipid composition can in principle occur at any preparation step due to nonideal mixing, lipid-surface interactions, etc. Here, we report a straightforward HPLC-ELSD method to quantify the lipid concentration and composition of liposomes and apply that method to study the preparation of simple cholesterol/POPC liposomes. We examine common liposome preparation steps, including vortexing during resuspension, lipid film hydration, extrusion, freeze-thaw, and sonication. We found that the resuspension step can play an outsized role in determining the lipid loss (up to ∼50% under seemingly rigorous procedures). The extrusion step yielded smaller lipid losses (∼10–20%). Freeze-thaw and sonication could both be employed to improve lipid yields. Hydration times up to 60 min and increasing cholesterol concentrations up to 50 mol % had little influence on lipid recovery. Fortunately, even conditions with large lipid loss did not substantially influence the target membrane composition, as long as the lipid mixture was below the cholesterol solubility limit. From our results, we identify best practices for producing maximum levels of lipid recovery and minimal changes to lipid composition during liposome preparation for cholesterol/POPC liposomes.

Changes in performance, cecal microflora counts and intestinal histology of Japanese quails fed diets containing different fibre sources

The purpose of this experiment was to investigate how various fiber sources impact the performance, microbial population, and intestinal histology of Japanese quail that was performed in a completely randomized design for 42 days. The dietary treatments involved a fiber-free corn-soybean meal-based diet (control, CTL), and CTL with added levels of sunflower hulls (SFH) and sugar beet pulp (SBP) (20 and 40 g kg-1). Body weight gain (BWG) and feed intake (FI) were recorded weekly. Carcass characteristics, cecal microbial population, blood variables and intestinal histology were measured on the 42 day of age. Adding 40 g kg-1 of SBP led to a significant decrease in body weight gain and an increase in the feed conversion ratio of birds from 1 to 21 days (P < 0.05). The relative weight of the gastrointestinal tract and gizzard increased significantly in birds that consumed SFH. Blood triglyceride concentration decreased with the inclusion of fiber in the diet. However, there was a notable increase in blood cholesterol concentration in the birds that were fed SBP (20 and 40 g kg-1) in comparison to those fed SFH (P < 0.05). The population of E. Coli in the cecum increased significantly in the birds that were fed 4 g kg-1 of SBP as opposed to those fed 20 and 40 g kg-1 of SFH (P < 0.05). The villus height of the jejunum in birds that were fed 20 g kg-1 and 40 g kg-1 of SFH demonstrated a significant increase in comparison to the other treatments (P < 0.05). In general, the findings of this research indicated that the inclusion of 40 g kg-1 of SBP in the diet had a negative impact on performance and other physiological parameters. However, the use of SFH and 20 g kg-1 of SBP yielded similar results to birds in the CTL, and in some cases, even better outcomes.

P-596 Impaired fatty acid desaturation in COV434 human granulosa cells leads to increased omega-6 phospholipid abundance

Abstract Study question How does fatty acid desaturation in COV434 granulosa cells influence their lipogenic pathway? Summary answer Suppressing fatty acid desaturation in human granulosa cells leads to a depletion of polyunsaturated phospholipids and an increase in omega-6 fatty acid-containing lipids. What is known already Maintaining the balance between saturated and unsaturated lipids in oocyte-surrounding granulosa cells is crucial to supporting oocyte maturation. We have shown the link between the de novo production of oleic acid through fatty acid desaturation and the main functional parameters of granulosa cells, such as cell proliferation and steroid production. A potential contributor to the observed pathophysiological outcomes lies in the dysregulation of cellular lipids as a consequence of insufficient intracellular oleic acid availability. This lipidomic study investigates the adaptive lipogenic response of granulosa cells to deficient fatty acid desaturation. Study design, size, duration We tested the influence of a chemical inhibitor against stearoyl-CoA desaturase 1 (SCD1), a major fatty acid desaturase enzyme, on cell membrane lipid packing, lipidome, and lipogenic-related genes in immortalized granulosa cells, COV434. We applied free oleic acid to conduct a rescue experiment. Three independent technical replicates were performed for each experimental condition, and mean values were compared among test conditions using one-way ANOVA analysis. Participants/materials, setting, methods The level of membrane fluidity was estimated using the membrane polarity-sensitive dye Laurdan. We used liquid chromatography–mass spectrometry to investigate the lipidome profiles. Data were obtained by calculating the peak areas of intact lipid species. We used quantitative PCR with TaqMan primers to assess the expression of mechanistic target of rapamycin (mTOR), diacylglycerol acyltransferase 1 (DGAT1), and fatty acid synthase (FASN), with 18S ribosomal RNA as the internal control. Main results and the role of chance Reduced oleic acid availability increased cholesterol content 1.2-fold (p = 0.007) and membrane packing, representing intended modifications in de novo desaturation. We used data comprising 459 lipid species. SCD1 inhibition caused a significant increase in dipalmitoyl phosphatidic acid 32:0 (7.4-fold increase vs. untreated controls, p &amp;lt; 0.001) and a substantial decrease in dioleoyl phosphatidic acid 36:2 (-38% reduction, p &amp;lt; 0.001). SCD1 inhibition also led to an elevation of omega-6 phospholipids (e.g., 38:4, 40:4), but conversely, a depletion of polyunsaturated lipids (e.g., 34:2, 36:2). In contrast, oleic acid supplementation resulted in an enrichment of oleic acid-containing lipids (e.g. 34:1, 36:1) but also caused a significant depletion of all polyunsaturated lipids, including omega-6 species (e.g., 20:3, 20:4). These findings indicate that the de novo generation of oleic acid intricately modulates polyunsaturated phospholipid metabolism, especially that of omega-6 phospholipids.Although the reductions in the expression levels of lipogenic-related genes due to SCDinhib treatment were not statistically significant, the addition of oleic acid significantly enhanced the suppression of FASN by 67% (p = 0.022). This suggests the presence of an interaction effect that potentiates the downregulation of the de novo fatty acid synthesis and diverts metabolic pathways away from omega-6 fatty acid production. Limitations, reasons for caution Examining lipidomic profile in isolated primary ovarian cells is crucial for a better understanding of their biological significance. Wider implications of the findings We identified that the role of de novo oleic acid generation in polyunsaturated phospholipid metabolism has significance for cellular physiology in oocyte-surrounding granulosa cells. Potential future applications include targeted metabolic interventions for reproductive disorders impacting oogenesis. Trial registration number not

Cholesterol and Cytokines: Molecular Links to Atherosclerosis and Carcinogenesis.

An increase of cholesterol concentration within the artery obstructs arterial blood flow once it deposits alongside the arterial wall. This results in atherosclerosis. Carcinogenesis causes a quicker clearance of vascular cholesterol to meet the demands of tumour cell development. Both illnesses have an increased concentration of pro-inflammatory cytokines in the blood. To search the comparative characteristics of cholesterol and pro-inflammatory cytokines in the pathogenesis of atherosclerosis and carcinogenesis, a comprehensive online survey using MEDLINE, Scopus, PubMed, and Google Scholar was conducted for relevant journals with key search term cholesterol and cytokines in atherosclerotic and cancerous patients. According to reports, hypercholesterolaemia related dyslipidemia causes atherosclerosis in blood arteries and hypercholesterolaemia in cell nucleus is a reason for developing carcinogenesis. It is also noted that pro-inflammatory cytokines are involved in both of the aforementioned pathogenesis. Changes in anti-inflammatory cytokines are only the characteristic features of each kind. Thus, Cholesterol and pro-inflammatory cytokines are intensely interlinked in the genesis of atherosclerotic and carcinogenic consequences.

Early Effects of Modern Radiotherapy for Lung Cancer on Endothelial Damage and Myocardial Fibrosis: A Prospective Single-Center Study.

Radiotherapy (RT) may have a cardiotoxic effect on the heart and cardiovascular system. Postulated mechanisms mediating these complications include vascular endothelium damage and myocardial fibrosis. The aim of our study was to assess endothelial damage and myocardial fibrosis in the early period after RT on the basis of cardiac biomarkers and in relation to the radiation dose applied to individual heart structures in patients treated for non-small-cell lung cancer. This single-center prospective study included consecutive patients with lung cancer (LC) who were referred for treatment with radiochemotherapy (study group) or chemotherapy (control group). The study protocol included performing an echocardiographic examination, a standard ECG examination, and collecting blood samples for laboratory tests before starting treatment for lung cancer in the first week after completing RT (after four cycles of chemotherapy in the control group) and after 12 weeks from the end of treatment. The study included 23 patients in the study group and 20 patients in the control group. Compared to the baseline values, there was a significant increase in total cholesterol concentration in the study group immediately after the end of RT, which persisted for three months after the end of therapy. After taking into account the use of statins in the analysis, it was found that an increase in total cholesterol concentration after oncological treatment was observed only among patients who did not use statins. Taking into account the assessment of myocardial fibrosis markers, there were no significant changes in the concentration of matrix metallopeptidase 9 (MMP-9) and tissue inhibitors of metalloproteinases 1 (TIMP-1) in the study group. In patients treated with radiochemotherapy, there was a significant increase in the concentration of intercellular adhesion molecule 1 (ICAM-1) immediately after RT, when compared to the baseline. After taking into account the use of statins, an increase in ICAM-1 concentration immediately after RT was observed only in patients who did not use statins. There was also a significant correlation between the radiation dose received by the left anterior descending coronary artery (LAD) and left circumferential coronary artery, and vascular cell adhesion protein 1 (VCAM-1) concentration measured at three months after the end of RT. Immediately after completion of radiotherapy, a significant increase in the level of ICAM-1 is observed indicating endothelial damage. The radiation dose to coronary arteries should be minimized, as it correlates with the concentration of VCAM-1. The use of statins may prevent the increase in total cholesterol and ICAM-1 concentration after irradiation for lung cancer; however, further studies designed for this specific purpose are necessary to confirm the effectiveness of statins in this area.

Study of the effect of anemia on lipid levels in some patients in the city of Mosul

Many scientific studies indicated that there is no close correlation between the malnutrition and weight gain (obesity) with iron deficiency, which leads to anemia. This study was designed to find the nature of the relationship between fat accumulations in adults with anemia.This study and tests were conducted in Mosul/Iraq, and random samples werecollected from adults in two clinical analysis laboratories accredited by the Department of Health. The samples were taken after fasting for 14 consecutive hours to check out the level of cholesterol, LDL, the lipase enzyme, and some chemical elements such as iron and copper, as well as measuring the HB% percentage. The results were analyzed statistically based on the t-test using SPSS statistical program, and a value of p&amp;#60;0.05 was considered a significant value within the results.The ages of the people targeted in this investigation were between (18-50) years, and after collecting the test results and analyzing them statistically, the samples were divided into three groups based on the severity of iron deficiency (anemia).The people included in the first group (group A) were with severe anemia, while people in the second group (group B) were for moderate to weak anemia, whereas the third group (group C) designed to be a control group (people who do not have anemia). A comparison was made between group A with the control group C, and the average cholesterolconcentration in group A appeared to be higher than that in group C,-With a significant difference, while the average lipase and LDL levels were slightly, higher in group A compared to the control group, but it was not significant. As for the chemical elements, the average Fe in group A was low, with a significant difference at p&amp;#60;0.05 compared with C, while the copper element, was low in the first group compared to the third group, but it was not significant. The average hemoglobin (Hb) concentration was different between the three mentioned groups, and it was low in the first group compared to the second and third group. There were a significant changes in Hb concentrations between group A compared to group C, while there was no significant difference between group B and group C.The results of the study, after comparing the groups, showed a significant increase in cholesterol concentration in people with all groups of anemia and non-significant differences based on statistical significances in the concentration of lipase and low-density lipoprotein in people with anemia compared to the control group. As in copper element, it was non-significantly low in people with anemia, in contrast, the concentration of iron was low and significant in those with anemia, which is the significant cause of anemia in adults. The study results may agree with previous studies and/ or different from other studies. Therefore, it is suggested that more experiments and research have to be done to find the physiological causes for the extent of the effects of anemia on fat concentrations.

Assessment of non-communicable disease risk factors, functional performance, and health-related quality of life in adults: a comparative analysis in low-resourced urban and rural areas of South Africa

BackgroundGlobally, disparities between non-communicable disease (NCD) risk factors, functional performance, and health-related quality of life (HRQoL) exist in people living in rural and low-resourced urban settings. Evidence of these health differences determined with objective NCD risk factors and functional performance measurements in South Africa, is scarce. Therefore, the study aimed to determine the differences in NCD risk factors, functional performance and HRQoL between rural and low-resourced urban areas.MethodsThe study recruited 311 adults (35–80 years) presenting with at least one NCD risk factor from low-resourced urban- (n = 183) and rural (n = 128) communities. Objective measurements of physical activity (PA) by means of combined heart rate and accelerometery, body composition employing skinfolds, peripheral lipid and glucose concentrations, blood pressure, functional performance indicators (handgrip, single leg stand, sit-to-stand, timed-up-and-go speed, predicted peak VO2 max); and HRQoL were measured according to standard procedures. Independent t-tests, Mann-Whitney U, and chi-square tests were performed to determine differences between the variables of low-resourced urban and rural settings.ResultsThe participants from the low-resourced urban setting were significantly older than the rural residents (59.1 ± 10.7 years vs. 52.8 ± 11.3 years; p = 0.001). NCD risk factors were significantly more prevalent in the low-resourced urban participants compared to rural participants, in particular for elevated systolic (85.8% vs. 62.5%; p = 0.001), and diastolic blood pressure (88.5% vs. 65.6%; p = 0.001), physical inactivity (95.9% vs. 87.7%; p = 0.026), increased cholesterol concentrations (22.1% vs. 8.7%; p = 0.002), and increased waist circumference (61.9% vs. 49.2%; p = 0.027). Low-resourced urban residents presented with a higher average body fat percentage (27.69% ± 7.65% vs. 12.23% ± 4.67%; p < 0.001), and lower moderate to vigorous PA levels (37.19 ± 49.55 [95% CI = 29.12–45.27] vs. 62.92 ± 60.43 min/week [95% CI = 47.95–77.90]; p = 0.003) compared to rural residents. Rural residents showed significantly better functional performance, including peak VO2 (23.99 ± 9.89 vs. 16.95 ± 7.64 ml/min/kg; p = 0.001) and single leg stand (right leg: 44.96 ± 18.47 vs. 20.87 ± 19.18 s; p = 0.001) as well as higher HRQoL for the physical (51.06 ± 8.14% vs. 45.62 ± 11.13%; p < 0.001) and mental (54.75 ± 8.24% vs. 48.91 ± 12.27%; p < 0.001) component scores compared to participants from the low-resourced urban areas.ConclusionNCD risk factors, functional performance, and HRQoL significantly differ in rural communities compared to low-resourced urban communities in South Africa. Urban areas’ most prevalent risk factors were elevated blood pressure, physical inactivity, and increased waist circumference. Participants from rural areas demonstrated significantly better functional performance, such as fitness and balance. HRQoL was better in rural settings than in urban settings. Future intervention programmes should be tailored for specific settings.

Nanoliposomal Encapsulation of Capparis spinosa Extract and Its Application in Jelly Formulation.

This research aimed to encapsulate the Capparis spinosa fruit extract to increase its stability for incorporation into food products such as jelly or jelly powder. After extraction, the nanoliposomes containing the extract were prepared in ratios of 60-0, 50-10, 40-20, and 30-30 lecithin-to-cholesterol. The effects of lecithin-to-cholesterol concentrations on the related parameters were then evaluated. The results showed that the average particle size was in the range of 95.05 to 164.25 nm, and with an increasing cholesterol concentration, the particle size of the nanoliposomes increased. The addition of cholesterol increased the zeta potential from -60.40 to -68.55 millivolt. Furthermore, cholesterol led to an increase in encapsulation efficiency, and even improved the stability of phenolic compounds loaded in nanoliposomes during storage time. Fourier transform infrared (FTIR) spectroscopy confirmed the successful loading of the extract. Field emission scanning electron microscopy (FE-SEM) analysis revealed nano-sized spherical and almost-elliptical liposomes. For jelly powders, the water solubility index ranged from 39.5 to 43.7% (p > 0.05), and the hygroscopicity values ranged between 1.22 and 9.36 g/100 g (p < 0.05). In conclusion, nanoencapsulated Capparis spinosa extract displayed improved stability and can be used in jelly preparation without any challenge or unfavorable perception.

Lipid loss and compositional change during preparation of liposomes by common biophysical methods.

Liposomes are widely used as model lipid membrane platforms in many fields, ranging from basic biophysical studies to drug delivery and biotechnology applications. Various methods exist to prepare liposomes, but common procedures include thin-film hydration followed by extrusion, freeze-thaw, and/or sonication. These procedures have the potential to produce liposomes at specific concentrations and membrane compositions, and researchers often assume that the concentration and composition of their liposomes are similar to, if not identical, to what would be expected if no lipid loss occurred during preparation. However, lipid loss and concomitant biasing of lipid composition can in principle occur at any preparation step due to nonideal mixing, lipid-surface interactions, etc. Here, we report a straightforward method using HPLC-ELSD to quantify the lipid concentration and membrane composition of liposomes, and apply that method to study the preparation of simple POPC/cholesterol liposomes. We examine many common steps in liposome formation, including vortexing during re-suspension, hydration of the lipid film, extrusion, freeze-thaw, sonication, and the percentage of cholesterol in the starting mixture. We found that the resuspension step can play an outsized role in determining the overall lipid loss (up to ~50% under seemingly rigorous procedures). The extrusion step yielded smaller lipid losses (~10-20%). Freeze-thaw and sonication could both be employed to improve lipid yields. Hydration times up to 60 minutes and increasing cholesterol concentrations up to 50 mole% had little influence on lipid recovery. Fortunately, even conditions with large lipid loss did not substantially influence the target membrane composition more than ~5% under the conditions we tested. From our results, we identify best practices for producing maximum levels of lipid recovery and minimal changes to lipid composition during liposome preparation protocols. We expect our results can be leveraged for improved preparation of model membranes by researchers in many fields.

The Influence of a High-Cholesterol Diet and Forced Training on Lipid Metabolism and Intestinal Microbiota in Male Wistar Rats.

Adequate experimental animal models play an important role in an objective assessment of the effectiveness of medicines and functional foods enriched with biologically active substances. The aim of our study was a comparative assessment of the effect of consumption of 1 or 2% cholesterol with and without regular (two times a week), moderate running exercise on the main biomarkers of lipid and cholesterol metabolism, as well as the intestinal microbiota of male Wistar rats. In experimental rats, a response of 39 indicators (body weight, food consumption, serum biomarkers, liver composition, and changes in intestinal microbiota) was revealed. Total serum cholesterol level increased 1.8 times in animals consuming cholesterol with a simultaneous increase in low-density lipoprotein cholesterol (2 times) and decrease in high-density lipoprotein cholesterol (1.3 times) levels compared to the control animals. These animals had 1.3 times increased liver weight, almost 5 times increased triglycerides level, and more than 6 times increased cholesterol content. There was a tendency towards a decrease in triglycerides levels against the background of running exercise. The consumption of cholesterol led to a predominance of the Bacteroides family, due to a decrease in F. prausnitzii (1.2 times) and bifidobacteria (1.3 times), as well as an increase in Escherichia family (1.2 times). The running exercise did not lead to the complete normalization of microbiota.

Effect of cholesterol-loaded cyclodextrin treatment on boar sperm cryopreservation.

This study investigated the efficacy of different concentrations of cholesterolloaded cyclodextrin (CLC) on cryopreservation in boar sperm quality. In this study, we treated boar sperm with different concentrations of CLC before freezing and analyzed the sperm cholesterol concentration, plasma membrane, acrosome integrity rate and total motility rate before and after freeze-thawing. We also investigated the levels of reactive oxygen species (ROS), malondialdehyde (MDA), adenosine triphosphate (ATP), and structural- and oxidative-damage related proteins in all groups after thawing. The results revealed that the cholesterol concentration of the CLC-treated groups was higher than that of the control group, both before freezing and after thawing (p<0.05). The plasma membrane integrity rate, acrosome integrity rate, and total motility rate of sperm were also enhanced after thawing in the CLC-treated group (all p<0.05). Moreover, ROS and MDA production and ATP loss were reduced in CLC-treated sperm during freezing and thawing (p<0.05). Finally, CLC pretreatment partially prevented the consumption of various proteins involved in metabolism including capping actin protein of muscle Z-line subunit beta (CAPZB), heat shock protein 90 alpha family class A member 1 (HSP90AA1) and phosphoglycerate mutase 2 (PGAM2) (p<0.05). The CLC treatment increased cholesterol concentration and decreased structural injury and oxidative damage during boar sperm freezing and thawing, improving the efficacy of sperm cryopreservation in boar.

Association of FoxP3&lt;sup&gt;+&lt;/sup&gt; T regulatory lymphocytes with epicardial adipose tissue thickness in patients with coronary heart disease

Background. Increase ofthe epicardial adipose tissue (EAT) thickness isassociated with development of inflammation and cardiovascular complications, however, there is no data on the relationship between EAT thickening and the number of immunosuppressive regulatory T lymphocytes.The aim. To study the number of circulating T regulatory lymphocytes and nuclear translocation ofthe FoxP3 transcription factor in patients with stable coronary heart disease (CHD) depending on the epicardial adipose tissue thickness.Materials and methods. We examined 30 patients with chronic stable CHD. The EAT thickness was measured by echocardiography. Patients were divided into groups depending on the presence and absence of EAT thickening above 5 mm (groups 1 and 2, respectively). Imaging flow cytometry was used to determine the number of T regulatory lymphocytes and the level of FoxP3 nuclear translocation. The concentration of cytokines and high sensitivity C-reactive protein (hsCRP) was determined using enzyme-linked immunosorbent assay in blood serum.Results. Anthropometric indicators of obesity and the severity of atherosclerosis were comparable between groups. In group 2, there was an increase in low-density lipoprotein cholesterol concentration (p = 0.043), ratio of low-density lipoprotein cholesterol tohigh-density lipoprotein cholesterol (p = 0.017) and the concentration ofhsCRP (p = 0.044) andIL-1β (p = 0.005), adecrease in the number and relative count of Tregulatory lymphocytes (p = 0.020 andp = 0.026, respectively), aswellas thenumber of cells withFoxP3 nuclear translocation (p = 0.018) compared togroup1. According tomultiple logistic regression, the concentration ofhsCRP, IL-1β and T regulatory lymphocytes relative count in total were the predictors of EAT thickening (accuracy 80 %; sensitivity 75 %; specificity 84,6 %; AUC = 0.89).Conclusions. Thickening of epicardial adipose tissue inpatients withcoronary heart disease is associated with a decrease in the number of T regulatory lymphocytes andFoxP3 nuclear translocation inthem in presence of comparable anthropometric parameters of obesity and the severity of coronary atherosclerosis.

Impacts of astaxanthin-enriched Paracoccus carotinifaciens on growth, immune responses, and reproduction performance of broodstock Nile tilapia during winter season.

The growth, immune response, and reproductive performance of broodstock of Nile tilapia (Oreochromis niloticus) under winter stress conditions were investigated the effects of supplementary diets with astaxanthin-enriched Paracoccus carotinifaciens. Throughout an eight-week period in the winter season, male and female tilapia were fed with diets containing different levels of P. carotinifaciens dietary supplementation: 0 g/kg (T1; control), 5 g/kg (T2), 10 g/kg (T3), and 20 g/kg (T4). Subsequently, a four-week mating system was implemented during the winter stress period. The results revealed that there were no significant differences observed in growth, hematological indices, and blood chemical profiles among all treatment groups for both male and female tilapia. However, a significant increase in cholesterol content was noted in both male and female tilapia fed with the T4 diet (p<0.05). The total carotenoid content in the muscle was evaluated, and significantly higher values were found in both male and female tilapia that fed T4 supplementation (p<0.05). Moreover, immunological parameters such as myeloperoxidase and antioxidant parameters in the liver including superoxide dismutase activity and catalase enzyme activity showed significant increases in tilapia fed with the T4 diet. The impact of P. carotinifaciens supplementation on broodstock tilapia indicated a significant increase in spermatozoa concentration in males and increased egg production in females after consumption of the T4 diet (p<0.05). Thus, this study highlighted that the presence of astaxanthin-enriched P. carotinifaciens in the diet of broodstock Nile tilapia can lead to the accumulation of carotenoids in their muscle tissue, improvement in antioxidant status, enhancement of immune function, and potential enhancement of reproductive capabilities, even under overwintering conditions.

The E3 ubiquitin ligase Itch regulates death receptor and cholesterol trafficking to affect TRAIL-mediated apoptosis

The activation of apoptosis signalling by TRAIL (TNF-related apoptosis-inducing ligand) through receptor binding is a fundamental mechanism of cell death induction and is often perturbed in cancer cells to enhance their cell survival and treatment resistance. Ubiquitination plays an important role in the regulation of TRAIL-mediated apoptosis, and here we investigate the role of the E3 ubiquitin ligase Itch in TRAIL-mediated apoptosis in oesophageal cancer cells. Knockdown of Itch expression results in resistance to TRAIL-induced apoptosis, caspase-8 activation, Bid cleavage and also promotes cisplatin resistance. Whilst the assembly of the death-inducing signalling complex (DISC) at the plasma membrane is not perturbed relative to the control, TRAIL-R2 is mis-localised in the Itch-knockdown cells. Further, we observe significant changes to mitochondrial morphology alongside an increased cholesterol content. Mitochondrial cholesterol is recognised as an important anti-apoptotic agent in cancer. Cells treated with a drug that increases mitochondrial cholesterol levels, U18666A, shows a protection from TRAIL-induced apoptosis, reduced caspase-8 activation, Bid cleavage and cisplatin resistance. We demonstrate that Itch knockdown cells are less sensitive to a Bcl-2 inhibitor, show impaired activation of Bax, cytochrome c release and an enhanced stability of the cholesterol transfer protein STARD1. We identify a novel protein complex composed of Itch, the mitochondrial protein VDAC2 and STARD1. We propose a mechanism where Itch regulates the stability of STARD1. An increase in STARD1 expression enhances cholesterol import to mitochondria, which inhibits Bax activation and cytochrome c release. Many cancer types display high mitochondrial cholesterol levels, and oesophageal adenocarcinoma tumours show a correlation between chemotherapy resistance and STARD1 expression which is supported by our findings. This establishes an important role for Itch in regulation of extrinsic and intrinsic apoptosis, mitochondrial cholesterol levels and provides insight to mechanisms that contribute to TRAIL, Bcl-2 inhibitor and cisplatin resistance in cancer cells.

Cuban policosanol improves high-density lipoprotein cholesterol efflux capacity in healthy Japanese subjects.

Policosanol supplementation has been reported to increase high-density lipoprotein (HDL)-cholesterol (HDL-C). However, the association between Cuban policosanol supplementation and HDL cholesterol efflux capacity (CEC), an important function of HDL, remains unclear. We performed a lipoprotein analysis investigating 32 Japanese healthy participants (placebo, n = 17 or policosanol supplementation for 12 weeks, n = 15) from a randomized Cuban policosanol clinical trial. First, HDL CEC and HDL-related factors were measured before and after policosanol supplementation. Then, through electron microscopy after ultracentrifugation and high-performance liquid chromatography, HDL morphology and subclass were analyzed, respectively. Finally, the effects of policosanol supplementation regarding HDL function, HDL-related factors, and HDL morphology/component were examined. Cuban policosanol considerably increased the HDL CEC and HDL-C and apolipoprotein A-I (ApoA-I) levels. Furthermore, policosanol supplementation led to larger HDL particles, increased cholesterol content in larger HDL particles, and reduced triglyceride content in smaller HDL particles. In participants with high baseline HDL-C levels, the policosanol effects for HDL CEC are observed. HDL CEC fluctuation induced by policosanol was highly associated with HDL-C and ApoA-I changes. In conclusion, for the first time, we demonstrated that policosanol supplementation increased the HDL CEC in healthy participants.

Determination of pore edge tension from the kinetics of rupture of giant unilamellar vesicles using the Arrhenius equation: effects of sugar concentration, surface charge and cholesterol.

The pore edge tension (Γ) of a membrane closely intertwines with membrane stability and plays a vital role in the mechanisms that facilitate membrane resealing following pore formation caused by electrical and mechanical tensions. We have explored a straightforward procedure to determine Γ by fitting the inverse of the tension-dependent logarithm of the rate constant of rupture of giant unilamellar vesicles (GUVs) using the Arrhenius equation. The GUVs were prepared using a combination of 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DOPG) and 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) in a physiological environment. The effects of sugar concentration, membrane surface charge density, and membrane cholesterol concentration on Γ have been investigated. The values of Γ increase with sugar concentration in the physiological buffer, measuring 9.6 ± 0.3, 10.4 ± 0.1, and 16.2 ± 0.1 pN for 40, 100, and 300 mM, respectively. A higher concentration of anionic lipids (70 mol% of DOPG) significantly reduces Γ. An increasing trend of Γ with cholesterol content was observed; specifically, the values of Γ were 11.9 ± 0.9, 13.9 ± 0.7, and 16.2 ± 0.4 pN for 15, 29, and 40 mol% cholesterol, respectively. Thus, the presence of higher anionic lipids in the bilayer led to a decrease in membrane stability. In contrast, the presence of higher sugar concentrations in the buffer and increased cholesterol concentration in the membranes enhanced membrane stability.