Non-esterified Cholesterol Research Articles

Physiological Effects of Soat1 Inactivation on Homeostasis of the Mouse Ocular Surface.

Soat1/SOAT1 have been previously reported to be critical for the biosynthesis of cholesteryl esters (CEs) in the mouse Meibomian glands (MGs) as the loss of function led to an arrest of CE production and a substantial accumulation of nonesterified cholesterol in the meibum, causing an increase in its melting temperature. The purpose of this study was to further investigate the role of Soat1 in meibogenesis and ocular surface physiology. The mouse ocular features of knockout Soat1-/- and wild type (WT) mice were studied using various ophthalmic and histological techniques, mouse lipidomes were monitored using liquid chromatography/mass spectrometry, whereas their transcriptomes were compared to characterize the effects of the mutation on the gene expression profiles in the MG and cornea. Soat1-/- mice displayed increased tear production and severe corneal abnormalities, such as corneal thinning, (neo)vascularization, ulceration, and opacification that progressed with aging. Transcriptomic analyses led to identification of a range of significantly disrupted pathways, which included general and specific lipid metabolism-related pathways, keratinization, angiogenesis/(neo)vascularization, muscle contraction, and several other pathways. In addition, histological and histochemical experiments revealed morphological changes in the MG, cornea, and conjunctiva in Soat1-/- mice. Notably, the mRNA microarray expression level of Soat1 in WT MGs (log2 17.5) was 1000 × of that in the mouse cornea (log2 7.5). These findings suggest a direct involvement of Soat1/SOAT1 in MGs in maintaining ocular surface homeostasis, in general, and corneal health, specifically.

Paraoxonase, haptoglobin, serum amyloid A, tumor necrosis factor and acetylcholinesterase levels in ewes with pregnancy toxemia

Objective: In this study were investigated serum paraoxonase, haptoglobin, tumor necrosis factor-α, acetylcholinesterase, serum amyloid A, beta-hydroxybutyric acid, nonesterified fatty acids, glucose, total protein, aspartate aminotransferase, gamma-glutamyl transferase, cholesterol and triglyceride in ewes with pregnancy toxemia Materials and Methods: This study consisted of 10 control and 10 groups with pregnancy toxemia, comprising 20 merino hybrid ewes aged between 2-6 years. Results: The analysis of blood serum samples revealed that serum amyloid A (SAA), haptoglobin (HPT), tumor necrosis factor-α (TNF), paraoxonase (PON1), acetylcholinesterase (ACHE), aspartate aminotransferase (AST), gamma-glutamyl transferaz (GGT) and cholesterol did not differ statistically significant between two groups although SAA, HPT, TNF, PON1, ACHE, AST, GGT levels were higher in ewes with pregnancy toxemia when compared to healthy ewes. Beta hydroxybutyric acid (BHBA) and nonesterified fatty acids (NEFA) (P<0.001), triglyceride, and total protein (P<0.01) increased that glucose (P<0.001) levels decreased in sheep with pregnancy toxemia compared with healthy ewes. Conclusion: Acetylcholinesterase, paraoxonase, haptoglobin, tumor necrosis factor-α, serum amyloid A concentration researched may prove beneficial laboratory findings diseases in sheep with pregnancy toxemia.

The effects of maternal calcium status during late gestation on lamb growth and survival in twin-bearing Merino ewes grazing pasture.

Our project aimed to investigate whether low maternal calcium status during late gestation decreased weight, growth and survival of lambs born to twin-bearing Merino ewes grazing pasture. On day 130 of gestation, approximately a 10 mL blood sample was collected from 108 twin-bearing Merino ewes. Blood pH and calcium ion concentration were measured in whole blood and serum was analysed for calcium, magnesium, phosphate, beta hydroxy butyrate, non-esterified fatty acids and cholesterol. Six to 18 h after birth and at 49 days of age, lamb live weight was recorded. Ewe serum calcium level at day 130 of gestation were divided into tertiles (1.58-2.13, 2.14-2.30, and 2.31-2.99 mmol/L) with ewes classified as either Low, Med or High (n = 36 per group). A general linear mixed model was used to analyse the data. Survival to 1, 2, 3 and 7 days of age was lower for lambs born to ewes in the Low calcium group compared with those born in the Med and High calcium groups (P < 0.05). At day 49 of age, lamb survival tended to be lower for lambs in the Low (73.6%) compared with Med (86.8%) and High (85.3%) calcium groups (P < 0.1). There were no differences in lamb body weight (P > 0.05). Calcium levels of ewes in the Low calcium group were, on average, indicative of subclinical hypocalcaemia, with this group of ewes also having the lowest lamb survival rate, suggesting that maternal subclinical hypocalcaemia reduces lamb survival. Further studies, particularly on improving maternal calcium levels, are warranted.

The Influence of Recombinant Bovine Somatotropin (rbST) on the Metabolic Profile and Milk Composition of Lactating Murrah Buffalo.

The use of recombinant bovine somatotropin (rbST) leads to an increase in variable amounts of milk production in buffalo, but there is a lack of information on the influence of rbST on their metabolism. This study looked at the effects of a single 500 mg dose of rbST on the lipid profile, liver and kidney function, and physical, chemical, and cellular constitution of milk in 14 buffalo over 14 days, from the 100th day of lactation, compared with 14 animals in a control group. From the first day after rbST, there was a rise in beta-hydroxybutyrate (β-HBO), possibly due to higher dry matter intake or the biotransformation of NEFA into β-HBO. The treatment did not influence blood glucose, non-esterified fatty acids (NEFAs), triglycerides, cholesterol, total protein, albumin, AST, GGT, bilirubin, urea, or creatinine levels. In 71.3% of the buffalo, there was a gradual increase in milk production, with the maximal response occurring in the first week followed by a gradual decrease, whilst in 21.4%, the increase in production occurred between 7 and 10 days. Only 7.1% of the animals did not respond. On the 3rd, 5th, 7th, and 10th days after treatment, an increase was found in daily milk production between the two groups equal to 1.04, 1.52, 1.42, and 1.06 L, respectively. In relative terms, this means an increase in milk production, respectively, of 15.1%, 21.0%, 19.8%, and 15.1%. The constitution of the milk showed no difference in the amounts of fat, lactose, total solids, or somatic cell count; however, on the third day after rbST administration, there was a decrease in protein. Notably, from the fifth day, the protein values showed no statistical difference. It can be concluded that the use of rbST in buffalo from the 100th day of lactation is metabolically safe since the treatment neither caused imbalances in fat metabolism nor overloaded the liver or renal function, and the changes in milk composition were transient and limited to a decrease in milk protein.

Evidence for hypoxia-induced dysregulated cholesterol homeostasis in preeclampsia: Insights into the mechanisms from human placental cells and tissues.

Preeclampsia (PE) poses a considerable risk to the long-term cardiovascular health of both mothers and their offspring due to a hypoxic environment in the placenta leading to reduced fetal oxygen supply. Cholesterol is vital for fetal development by influencing placental function. Recent findings suggest an association between hypoxia, disturbed cholesterol homeostasis, and PE. This study investigates the influence of hypoxia on placental cholesterol homeostasis. Using primary human trophoblast cells and placentae from women with PE, various aspects of cholesterol homeostasis were examined under hypoxic and hypoxia/reoxygenation (H/R) conditions. Under hypoxia and H/R, intracellular total and non-esterified cholesterol levels were significantly increased. This coincided with an upregulation of HMG-CoA-reductase and HMG-CoA-synthase (key genes regulating cholesterol biosynthesis), and a decrease in acetyl-CoA-acetyltransferase-1 (ACAT1), which mediates cholesterol esterification. Hypoxia and H/R also increased the intracellular levels of reactive oxygen species and elevated the expression of hypoxia-inducible factor (HIF)-2α and sterol-regulatory-element-binding-protein (SREBP) transcription factors. Additionally, exposure of trophoblasts to hypoxia and H/R resulted in enhanced cholesterol efflux to maternal and fetal serum. This was accompanied by an increased expression of proteins involved in cholesterol transport such as the scavenger receptor class B type I (SR-BI) and the ATP-binding cassette transporter G1 (ABCG1). Despite these metabolic alterations, mitogen-activated-protein-kinase (MAPK) signaling, a key regulator of cholesterol homeostasis, was largely unaffected. Our findings indicate dysregulation of cholesterol homeostasis at multiple metabolic points in both the trophoblast hypoxia model and placentae from women with PE. The increased cholesterol efflux and intracellular accumulation of non-esterified cholesterol may have critical implications for both the mother and the fetus during pregnancy, potentially contributing to an elevated cardiovascular risk later in life.

Some biochemical parameters and acute phase proteins in goats with pregnancy toxemia

The present study investigated the levels of acetylcholinesterase, tumor necrosis factor-α (TNF), paraoxonase (PON1), haptoglobin (HPT), serum amyloid A (SAA), beta-hydroxybutyric acid (BHBA), non-esterified fatty acid (NEFA), glucose, total protein, cholesterol, and triglyceride in Saanen goats with pregnancy toxemia. The animal material included Saanen goats aged 2-5 years. The study was conducted on a total of 20 Saanen goats: 10 pregnant and 10 healthy animals. After clinical examination, 10 goats with BHBA levels of more than 1.5 mmol/L of blood were included in the pregnancy toxemia group, and 10 healthy goats with BHBA levels of less than 1 mmol/L were included in the control group. There were no significant differences between the total protein, cholesterol, and triglyceride levels in the two groups. It was determined that, while the glucose level was low in the goats with pregnancy toxemia, BHBA, NEFA, acetylcholinesterase, tumor necrosis factor-α, paraoxonase, serum amyloid A, and haptoglobin levels were higher than they were in the healthy goats. In conclusion, it appears that ACHE, TNFa, PON, SAA, and HPT may be useful as additional indicators in the diagnosis of pregnancy toxemia and in the development of methods for its prevention.

Causal role of lipid metabolome on the risk of ischemic stroke, its etiological subtypes, and long-term outcome: A Mendelian randomization study

Background and aimsThe lipid profile is consistently associated with coronary artery disease (CAD) and ischemic stroke (IS). However, the lipoprotein subfractions have not been deeply explored in stroke subtypes, especially in IS outcome. MethodsWe performed two-sample Mendelian randomization (MR) analysis using 92 lipid traits measured by nuclear magnetic resonance in 115,000 subjects from the UK Biobank. Data for genetic associations with IS, its subtypes, and long-term outcome (LTO) were obtained from three cohorts of European ancestry: GIGASTROKE (73,652 cases, 1,234,808 controls), GODS (n = 1791) and GISCOME (n = 6165). Results obtained using CARDIoGRAMPlusC4D were used to identify differences with CAD. ResultsGenetically determined low concentration of medium high-density lipoprotein (HDL) particles (odds ratio (OR) = 0.92, 95% CI 0.88–0.96; p = 3.6 × 10−4) and its cholesterol content (OR = 0.92, 95% CI 0.88–0.96; p = 1.9 × 10−4) showed causal associations with an increased risk of stroke. Genetic predisposition to high apolipoprotein (apo)B to apoA-I ratio was causally associated with an increased risk of IS (OR = 1.12, 95% CI 1.06–1.18, p = 1.1 × 10−4), and a highly suggestive association was found between non-esterified cholesterol in low-density lipoprotein (LDL) and increased risk of atherothrombotic stroke (LAS) (OR = 1.35, 95% CI 1.10–1.66; p = 4.0 × 10−3). Low cholesterol in small and medium LDL was suggestively associated with poor LTO. ConclusionsOur results support that low medium HDL concentration was causally associated with an increased stroke risk, while high levels of non-esterified cholesterol in LDL were suggestively associated with an increased risk of LAS and with a better LTO.

Influence of Heat Stress on Body Temperatures Measured by Infrared Thermography, Blood Metabolic Parameters and Its Correlation in Sheep.

The aim of this research is to examine the influence of heat stress (HS) on body temperature (BT) measured rectally (RT) or by infrared thermography (IRT) of the nose (NT), eye (ET), leg (LT) and abdominal (AT) regions in intensively and extensively breed sheep and to detect a correlation between body temperature and metabolic response in sheep. A total of 33 Wurttemberg × Sjenica Pramenka sheep breeds were examined, 17 ewes were from outdoors and 16 were from indoor housing systems during three experimental periods (thermoneutral period, severe HS and moderate HS). Sheep under HS have a higher BT, and the magnitude of BT measured by infrared thermography (IRT) was higher than RT. LT and AT showed positive linear correlations with the temperature-humidity index (THI), while other ways of measuring BT did not give statistically significant correlations. Sheep under HS showed higher cortisol, insulin, total protein, albumin, urea, creatinine, bilirubin, aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transferase, alkaline phosphatase, lactate dehydrogenase, creatine kinase and index of insulin resistance, with lower values of triiodothyronine (T3), thyroxine (T4), non-esterified fatty acids, beta-hydroxybutyrate (BHB), glucose, calcium, inorganic phosphates, magnesium and cholesterol. BT and metabolic response were different in the function of the housing method of sheep. LT and AT showed a significant correlation with almost all blood parameters, and the strongest connections were made with T3, T4, BHB and the revised quantitative insulin sensitivity check index of insulin resistance. The abdomen and legs are good thermal windows because LT and AT are good summative responses to external ambient THI and internal metabolic changes in sheep under heat stress.

Effect of Parity, Body Condition Score at Calving, and Milk Yield on the Metabolic Profile of Gyr Cows in the Transition Period.

This study aimed to evaluate the effects of parity, body condition score (BCS) at calving, and milk yield on the metabolic profile of Gyr (Zebu) cows. Healthy cows in late pregnancy were grouped according to parity (primiparous, biparous, and multiparous); to BCS scale at calving (high-HBCS and normal-NBCS); and to milk yield (high-HP and moderate-MP production). BCS was assessed, and blood samples were collected on -21, -7, 0, 7, 21, and 42 days relative to parturition. The concentrations of non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), cholesterol, glucose, total protein (TP), albumin, total calcium (Ca), phosphorus (P), and magnesium (Mg); and activities of aspartate aminotransferase and gamma-glutamyltransferase were measured. Data were analyzed by two-way repeated measures ANOVA. The frequencies of high lipomobilization, subclinical ketosis, subclinical hypocalcemia (SCH), and the occurrence of diseases during early lactation were established. Regardless of grouping, NEFA, BHB, and cholesterol increased during early lactation; glucose showed higher values at calving; TP and albumin were higher at 21 and 42 DIM; and Ca, P, and Mg were lower at calving. Parity had little effect on the metabolic profile, HBCS did not differ from NBCS cows, and HP did not differ from MP cows in most metabolites. High lipomobilization in early lactation and SCH at calving were the most common imbalances but were not related to postpartum diseases. High-yielding Gyr cows have a balanced metabolic profile during the transition period, with few biologically relevant effects of parity, BCS at parturition, or milk yielded.

Tumor Lipid Signatures Are Descriptive of Acquisition of Therapy Resistance in an Endocrine-Related Breast Cancer Mouse Model.

The lipid metabolism adaptations of estrogen and progesterone receptor-positive breast cancer tumors from a mouse syngeneic model are investigated in relation to differences across the transition from hormone-dependent (HD) to hormone-independent (HI) tumor growth and the acquisition of endocrine therapy (ET) resistance (HIR tumors). Results are articulated with reported polar metabolome results to complete a metabolic picture of the above transitions and suggest markers of tumor progression and aggressiveness. Untargeted nuclear magnetic resonance metabolomics was used to analyze tumor and mammary tissue lipid extracts. Tumor progression (HD-HI-HIR) was accompanied by increased nonesterified cholesterol forms and phospholipids (phosphatidylcholine, phosphatidylethanolamine, sphingomyelins, and plasmalogens) and decreased relative contents of triglycerides and fatty acids. Predominating fatty acids became shorter and more saturated on average. These results were consistent with gradually more activated cholesterol synthesis, β-oxidation, and phospholipid biosynthesis to sustain tumor growth, as well as an increase in cholesterol (possibly oxysterol) forms. Particular compound levels and ratios were identified as potential endocrine tumor HD-HI-HIR progression markers, supporting new hypotheses to explain acquired ET resistance.

Associations between the detailed milk mineral profile, milk composition, and metabolic status in Holstein cows

The causes of variation in the milk mineral profile of dairy cattle during the first phase of lactation were studied under the hypothesis that the milk mineral profile partially reflects the animals' metabolic status. Correlations between the minerals and the main milk constituents (i.e., protein, fat, and lactose percentages), and their associations with the cows' metabolic status indicators were explored. The metabolic status indicators (MET) that we used were blood energy-protein metabolites [nonesterified fatty acids, β-hydroxybutyrate (BHB), glucose, cholesterol, creatinine, and urea], and liver ultrasound measurements (predicted triacylglycerol liver content, portal vein area, portal vein diameter and liver depth). Milk and blood samples, and ultrasound measurements were taken from 295 Holstein cows belonging to 2 herds and in the first 120 d in milk (DIM). Milk mineral contents were determined by ICP-OES; these were considered the response variable and analyzed through a mixed model which included DIM, parity, milk yield, and MET as fixed effects, and the herd/date as a random effect. The MET traits were divided in tertiles. The results showed that milk protein was positively associated with body condition score (BCS) and glucose, and negatively associated with BHB blood content; milk fat was positively associated with BHB content; milk lactose was positively associated with BCS; and Ca, P, K and S were the minerals with the greatest number of associations with the cows' energy indicators, particularly BCS, predicted triacylglycerol liver content, glucose, BHB and urea. We conclude that the protein, fat, lactose, and mineral contents of milk partially reflect the metabolic adaptation of cows during lactation and within 120 DIM. Variations in the milk mineral profile were consistent with changes in the major milk constituents and the metabolic status of cows.

A novel detection method for neuronal death indicates abnormalities in intracellular membranous components in neuronal cells that underwent delayed death.

Acute neuronal degeneration is always preceded under the light and electron microscopes by a stage called microvacuolation, which is characterized by a finely vacuolar alteration in the cytoplasm of the neurons destined to death. In this study, we reported a method for detecting neuronal death using two membrane-bound dyes, rhodamine R6 and DiOC6(3), which may be associated with the so-called microvacuolation. This new method produced a spatiotemporally similar staining pattern to Fluoro-Jade B in kainic acid-damaged brains in mice. Further experiments showed that increased staining of rhodamine R6 and DiOC6(3) was observed only in degenerated neurons, but not in glia, erythrocytes, or meninges. Different from Fluoro-Jade-related dyes, rhodamine R6 and DiOC6(3) staining is highly sensitive to solvent extraction and detergent exposure. Staining with Nile red for phospholipids and filipin III for non-esterified cholesterol supports that the increased staining of rhodamine R6 and DiOC6(3) might be associated with increased levels of phospholipids and free cholesterol in the perinuclear cytoplasm of damaged neurons. In addition to kainic acid-injected neuronal death, rhodamine R6 and DiOC6(3) were similarly useful for detecting neuronal death in ischemic models either in vivo or in vitro. As far as we know, the staining with rhodamine R6 or DiOC6(3) is one of a few histochemical methods for detecting neuronal death whose target molecules have been well defined and therefore may be useful for explaining experimental results as well as exploring the mechanisms of neuronal death.

Metabolic indicators and their relationship with reproductive efficiency in dairy cows in the high tropics

The specialized herds of the high tropics of Nariño are characterized by semi-intensive breeding models, that is, feeding based on grazing and supplementation. The objective was to analyze the milk production of grazing Holstein cows under conditions of the high tropics, taking into account the energetic and protein metabolites, and their relationship with the dry matter intake (CMS/Kg/d), the body condition (BC), and the reproductive variables. Three periods of lactation were evaluated, thus, between 4-10 days (P1), 45-55 days (P2), and between 185-200 days (P3). The investigation was carried out in four dairy production farms. The forage supply consisted of a mixture of Lolium hybridum, Cenchrus clandestinus Hoschst ex Chiov, Holcus lanatus and Trifolium repens, plus commercial concentrate. A total of 24 Holstein animals (six per productive system) from three births onwards were used, from which blood samples were obtained to determine: non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), triglycerides, cholesterol, glucose, total protein (PT), and blood urea nitrogen (BUN). The results showed that milk production during P1 is influenced by CC and body weight (Kg) by +0.04 liters of milk per unit of change in this variable. In P2, the positive incidence variables were PT, BUN, glucose, NEFA, BHB, triglycerides, and cholesterol, and in P3, the increase in CMS made it possible to show that as lactation progresses in more than 180 days, the animals substantially improve their body condition; likewise the NEFA and BHB indicators are related to a better reproductive response.

The Effect of the Feeding System on Fat Deposition in Yak Subcutaneous Fat.

Fat deposition is very important to the growth and reproduction of yaks. In this study, the effect of the feeding system on fat deposition in yaks was explored by transcriptomics and lipidomics. The thickness of the subcutaneous fat in yaks under stall (SF) and graze feeding (GF) was evaluated. The transcriptomes and lipidomes of the subcutaneous fat in yaks under different feeding systems were detected by RNA-sequencing (RNA-Seq) and non-targeted lipidomics based on ultrahigh-phase liquid chromatography tandem mass spectrometry (UHPLC-MS), respectively. The differences in lipid metabolism were explored, and the function of differentially expressed genes (DEGs) was evaluated by gene ontology (GO) and Kyoto encyclopedia of genes and genome (KEGG) analysis. Compared with GF yaks, SF yaks possessed stronger fat deposition capacity. The abundance of 12 triglycerides (TGs), 3 phosphatidylethanolamines (PEs), 3 diglycerides (DGs), 2 sphingomyelins (SMs) and 1 phosphatidylcholine (PC) in the subcutaneous fat of SF and GF yaks was significantly different. Under the mediation of the cGMP-PKG signaling pathway, the blood volume of SF and GF yaks may be different, which resulted in the different concentrations of precursors for fat deposition, including non-esterified fatty acid (NEFA), glucose (GLU), TG and cholesterol (CH). The metabolism of C16:0, C16:1, C17:0, C18:0, C18:1, C18:2 and C18:3 in yak subcutaneous fat was mainly realized under the regulation of the INSIG1, ACACA, FASN, ELOVL6 and SCD genes, and TG synthesis was regulated by the AGPAT2 and DGAT2 genes. This study will provide a theoretical basis for yak genetic breeding and healthy feeding.

Lipid Profile of the Freshwater Pearl Mussel Margaritifera margaritifera Inhabiting Different Biotopes of the Lake-River System of the Kem River, White Sea Basin

The freshwater pearl mussel Margaritifera margaritifera is considered to be the most rapidly declining species of freshwater bivalve, whereas its colonies in rivers of the White Sea Basin remain the most numerous in the world. The lipid profiles of mantle, muscle (foot), and digestive gland tissues of the freshwater pearl mussel from the Kem, Ukhta, and Vozhma Rivers in autumn (end of September, early October) were studied using high performance thin-layer chromatography. The highest total lipid [TL] content was found in the digestive gland. Cholesterol esters, non-esterified fatty acids, phospholipids, and cholesterol were the dominant lipids in all studied tissues. The reduced triacylglycerol content in the mussels was associated with its utilization during the spawning period. The colony of the freshwater pearl mussel inhabiting the Vozhma River was distinguished by higher TL content in the mantle and digestive gland. Data on the size-age characteristics of mollusks from the Kem, Ukhta, and Vozhma Rivers and the relationship between the structural and storage lipid content and size-age parameters are discussed. The results are important for different conservation strategies of endangered species, such as the freshwater pearl mussel, especially in ecological monitoring based on evaluation of the physiological and biochemical state of mollusks and rare natural colonies.

A randomized controlled trial investigating the effect of transport duration and age at transport on surplus dairy calves: Part II. Impact on hematological variables

Surplus dairy calves often arrive at veal and dairy-beef rearing facilities with health and blood metabolite level abnormalities, which can affect their welfare and performance, predisposing them to future health challenges. The objective of this randomized controlled trial was to investigate the effects of transport duration and age at the time of transport on blood parameters in surplus dairy calves following 6, 12, or 16 h of continuous road transportation. All surplus calves from 5 commercial dairy farms in Ontario were enrolled and examined daily before transport (n = 175). On the day of transportation, calves were weighed, blood sampled, and randomly assigned to 6, 12, or 16 h of transportation. Blood samples were then collected immediately after transportation, as well as 24, 48, and 72 h thereafter. Serum was analyzed at a provincial diagnostic laboratory for nonesterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), creatine kinase (CK), cholesterol, and haptoglobin. In addition, blood gas and electrolyte values were also assessed at the time of sample collection. Mixed models with repeated measures were used to assess the effects of transport duration, breed, sex, transfer of passive immunity status, weight before transportation, and age at transportation on blood parameters. Immediately following transportation, NEFA and BHBA were greater for calves transported for 12 h (Δ = 0.22 mmol/L NEFA, 95% CI = 0.15 to 0.30; Δ = 0.04 mmol/L BHBA, 95% CI = 0.02 to 0.06) and 16 h (Δ = 0.35 mmol/L NEFA, 95% CI = 0.27 to 0.42; Δ = 0.10 mmol/L BHBA, 95% CI = 0.08 to 0.11) compared with calves transported for 6 h. Glucose was lower immediately following transportation in calves transported for 16 h compared with 6 h (Δ = -15.54 mg/dL, 95% CI = -21.54 to -9.54). In addition, pH and HCO3- were lower in calves transported for 12 (Δ = -0.09 pH, 95% CI = -0.13 to -0.05; Δ = -1.59 mmol/L HCO3-, 95% CI = -2.61 to -0.56) and 16 h (Δ = -0.07 pH, 95% CI = -0.12 to -0.03; Δ = -1.95 mmol/L HCO3-, 95% CI = -2.95 to -0.95) compared with calves transported for 6 h. Calves transported between 15 and 19 d of age had a higher concentration of cholesterol and CK (Δ = 0.27 mmol/L cholesterol; 37.18 U/L CK) compared with 2- to 6-d-old calves, and calves 12 to 14 d old had greater reduction in HCO3- (Δ = -0.92 mmol/L) compared with 2- to 6-d-old calves. These findings show that transporting calves for long distances results in lower glucose concentration and suboptimal energy status, and that this effect varies based on the calf's age.

The Antifungal Antibiotic Filipin as a Diagnostic Tool of Cholesterol Alterations in Lysosomal Storage Diseases and Neurodegenerative Disorders.

Cholesterol is the most considerable member of a family of polycyclic compounds understood as sterols, and represents an amphipathic molecule, such as phospholipids, with the polar hydroxyl group located in position 3 and the rest of the molecule is completely hydrophobic. In cells, it is usually present as free, unesterified cholesterol, or as esterified cholesterol, in which the hydroxyl group binds to a carboxylic acid and thus generates an apolar molecule. Filipin is a naturally fluorescent antibiotic that exerts a primary antifungal effect with low antibacterial activity, interfering with the sterol stabilization of the phospholipid layers and favoring membrane leakage. This polyene macrolide antibiotic does not bind to esterified sterols, but only to non-esterified cholesterol, and it is commonly used as a marker to label and quantify free cholesterol in cells and tissues. Several lines of evidence have indicated that filipin staining could be a good diagnostic tool for the cholesterol alterations present in neurodegenerative (e.g., Alzheimer's Disease and Huntington Disease) and lysosomal storage diseases (e.g., Niemann Pick type C Disease and GM1 gangliosidosis). Here, we have discussed the uses and applications of this fluorescent molecule in lipid storage diseases and neurodegenerative disorders, exploring not only the diagnostic strength of filipin staining, but also its limitations, which over the years have led to the development of new diagnostic tools to combine with filipin approach.

Molecular profile and peripheral markers of neurodegeneration in patients with Niemann-Pick type C: Decrease in Plasminogen Activator Inhibitor type 1 and Platelet-Derived Growth Factor type AA

Niemann-Pick type C1 (NPC1) is a fatal inherited disease, caused by pathogenic variants in NPC1 gene, which leads to intracellular accumulation of non-esterified cholesterol and glycosphingolipids. This accumulation leads to a wide range of clinical manifestations, including neurological and cognitive impairment as well as psychiatric disorders. The pathophysiology of cerebral damage involves loss of Purkinje cells, synaptic disturbance, and demyelination. Miglustat, a reversible inhibitor of glucosylceramide synthase, is an approved treatment for NPC1 and can slow neurological damage. The aim of this study was to assess the levels of peripheric neurodegeneration biomarkers of NPC1 patients, namely brain-derived neurotrophic factor (BDNF), platelet-derived growth factors (PDGF-AA and PDGF-AB/BB), neural cell adhesion molecule (NCAM), PAI-1 Total and Cathepsin-D, as well as the levels of cholestane-3β,5α,6β-triol (3β,5α,6β-triol), a biomarker for NPC1. Molecular analysis of the NPC1 patients under study was performed by next generation sequencing (NGS) in cultured fibroblasts. We observed that NPC1 patients treated with miglustat have a significant decrease in PAI-1 total and PDGF-AA concentrations, and no alteration in BDNF, NCAM, PDGF-AB/BB and Cathepsin D. We also found that NPC1 patients treated with miglustat have normalized levels of 3β,5α,6β-triol. The molecular analysis showed four described mutations, and for two patients was not possible to identify the second mutated allele. Our results indicate that the decrease of PAI-1 and PDGF-AA in NPC1 patients could be involved in the pathophysiology of this disease. This is the first work to analyze those plasmatic markers of neurodegenerative processes in NPC1 patients.

Variations of some adipokines, pro-inflammatory cytokines, oxidative stress biomarkers, and energy characteristics during the transition period in dairy cows.

Limited information exists about the relationship of adipose tissue with inflammation, oxidative stress, and energy metabolism during the transition period in dairy cows. The objective of this study was to assess the changes and relation of some adipokines, cytokines, oxidative biomarkers, and serum biochemical parameters related to energy balance (EB) in cows during the transition period. Thirty multiparous Holstein cows were selected based on estimated parturition date, and blood samples were collected from jugular vein on one-week prepartum and one and three weeks postpartum and used to measure the parameters. The serum levels of beta-hydroxybutyric acid (BHB), non-esterified fatty acid, cholesterol, high-density lipoprotein (HDL), aspartate aminotransferase, and total antioxidant capacity increased significantly, and glucose, urea, triglyceride (TG), and low-density lipoprotein (LDL) decreased significantly after parturition. The serum values of adiponectin, resistin, leptin, and cytokines including interleukin 6 (IL-6) and tumor necrosis factor (TNF)-α were not changed significantly during the experiment. The results of the Pearson correlation revealed a significant negative correlation between BHB with glucose, albumin, cholesterol, HDL, LDL, and a positive correlation with TG and malondialdehyde. Also, there was a significant direct correlation between insulin and leptin, adiponectin, resistin, IL-6 and TNF-α in the whole experiment period. These emphasize the difficulty of dairy cows to manage the energy requirements during the transition period. It can be stated that adipokines and cytokines may have an essential role in the metabolic status in this period, and control of their production and, or secretion could be helpful in EB during the transition period.

Differential distribution of cholesterol pools across arteries under high-cholesterol diet

Excessive cholesterol constitutes a major risk factor for vascular disease. Within cells, cholesterol is distributed in detergent-sensitive and detergent-resistant fractions, with the largest amount of cholesterol residing in cellular membranes. We set out to determine whether various arteries differ in their ability to accumulate esterified and non-esterified cholesterol in detergent-sensitive versus detergent-resistant fractions throughout the course of a high-cholesterol diet. Male Sprague-Dawley rats were placed on 2 % cholesterol diet while a control group was receiving iso-caloric standard chow. Liver, aorta, and pulmonary, mesenteric, and cerebral arteries were collected at 2–6, 8–12, 14–18, and 20–24 weeks from the start of high-cholesterol diet. After fraction separation, esterified and free non-esterified cholesterol levels were measured. In all arteries, largest cholesterol amounts were present in detergent-sensitive fractions in the non-esterified form. Overall, cholesterol in aorta and cerebral arteries was elevated during 14–18 weeks of high-cholesterol diet. Cerebral arteries also exhibited increase in esterified cholesterol within detergent-sensitive domains, as well as increase in cholesterol level in the detergent-resistant fraction at earlier time-points of diet. Pulmonary artery and mesenteric artery were largely resistant to cholesterol accumulation. Quantitative polymerase chain reaction (qPCR) analysis revealed up-regulation of low-density lipoprotein receptor (Ldlr) and low-density lipoprotein receptor-related protein 1 (Lrp1) gene expression in cerebral arteries when compared to mesenteric and pulmonary arteries, respectively. In summary, we unveiled the differential ability of arteries to accumulate cholesterol over the course of a high-cholesterol diet. The differential accumulation of cholesterol seems to correlate with the up-regulated gene expression of proteins responsible for cholesterol uptake.