Low-density Lipoprotein Protein Research Articles

Effect of Succinylation on Oxidation-Aggregation of Low-Density Lipoprotein and Formation of Off-Flavors in Heated Egg Yolks.

This study examined the effect of succinylation on protein oxidation-aggregation and the formation of off-flavors in heated egg yolks (EYs). The sensory evaluation, content of volatile compounds, stability of low-density lipoprotein (LDL) particles, and oxidation of lipid and protein at six levels of succinylated EY (0.25%, 0.5%, 1%, 2%, 5%, and 10%, w/w) were determined. The results showed that the succinylated thermal EY's concentration of volatiles and off-flavors was reduced. Oil exudation and lipid and protein oxidation decreased with the improved succinylation degree. Succinylation also reduced the LDL particle size and changed the secondary structure (decreased the β-sheets and increased the α-helices) of protein in LDL particles. Meanwhile, succinylation could effectively change the thermal oxidation-aggregation of LDL protein by introducing succinyl groups with negative charges, thus increasing the stability of LDL particles in succinylated EY during heating. These results further revealed the relationship between the oxidation-aggregation of LDL and the formation of off-flavors in heated EY. These results also help improve the flavor quality of heat-treated EY and expands the application scope of egg products.

Proatherogenic proteomic profile of LDL isolated from plasma of patients with diabetes mellitus: immunological aspects

Atherosclerosis is a vascular disease, which is based on a chronic inflammatory process, including complex interactions between blood components, as well as the cells that form the vessel wall, and lipid metabolism in general. Low-density lipoproteins (LDL) play a key role in pathogenesis of atherosclerosis. When the permeability of the endothelial layer of the vessel is impaired, LDL can penetrate into the intramural space and lead to excessive cholesterol accumulation by the cells from the intima subendothelial layer, leading to the onset of the inflammatory process and to the formation of foam cells, the main morphological component of the atherosclerotic plaque. However, the level of LDL in the patient’s blood plasma alone is not a key indicator of the development of atherosclerosis. Numerous studies point out the role of oxidized modifications of LDL (oxLDL) in the increased local accumulation of cholesterol in vascular wall cells, however, recent works have shown conflicting results regarding the role of oxLDL in the development of atherosclerosis. We hypothesize that other components of LDL may also influence the progression of atherosclerosis. It is a well-known fact that patients with diabetes mellitus (DM) suffer from cardiovascular diseases, in particular atherosclerosis, more often than patients without diagnosed DM and other autoimmune diseases, and the disease progresses faster. The purpose of this study was to identify risk biomarkers in LDL groups that indicate the relationship of the immune system with the development of atherosclerosis in such patients. LDL was isolated from patients and healthy donors using continuous ultracentrifugation with solutions of different densities, and LDL protein profile samples were measured using gas chromatography-mass spectrometry. We found 9 proteins that had a statistically significant difference between the samples (DM and control). In the diabetes group, the content of the antimicrobial peptide cathelicidin and lipopolysaccharide-binding protein was almost 2 times higher compared to the control. These proteins may be involved in the development of inflammation, leading to the progression of atherosclerosis. At the same time, a decrease in immunoglobulins and complement components (C9 and Complement C1s subcomponent) associated with LDL may contribute to the development of atherosclerosis.

Effect oral administration of saviotan feed on body temperature, HDL, LDL and total protein in Ulu chicks

Currently, the using of tannins as secondary metabolites and active compounds is rising in animal production. The functions of tannins especially extracted from chestnut wood, namely Saviotan Feed (SF) in poultry, especially in Ulu chickens, have not yet been fully determined. The aim of this study was to determine the effect of orally administered SF on some plasma metabolites, including body temperature, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and total protein (TP) concentration. In the study, 40 unsexed Ulu chicks aged 7 days were divided into four groups with different doses orally administered of SF (0%, 0.1%, 0.2%, and 0.3%). The body temperature of the chick was measured at 30, 60, and 120 min. At 120 min after oral administration, the blood samples were collected through the jugular vein, then centrifuges, and then plasma metabolites were analyzed. The results showed that no significant effect of SF on body temperature, LDL, and TP. However, it has a significant effect in increasing HDL concentration, especially SF at low levels. In conclusion, oral administration of SF increased HDL concentration, which is good for supporting health.

Identification of Glycoxidative Lesion in Isolated Low-Density Lipoproteins from Diabetes Mellitus Subjects.

Methylglyoxal (MG) is a precursor for advanced glycation end-products (AGEs), which have a significant role in diabetes. The present study is designed to probe the immunological response of native and glycated low-density lipoprotein (LDL) in experimental animals. The second part of this study is to probe glycoxidative lesion detection in low-density lipoproteins (LDL) in diabetes subjects with varying disease duration. The neo-epitopes attributed to glycation-induced glycoxidative lesion of LDL in DM patients' plasma were, analyzed by binding of native and MG-modified LDL immunized animal sera antibodies using an immunochemical assay. The plasma purified human LDL glycation with MG, which instigated modification in LDL. Further, the NewZealand-White rabbits were infused with unmodified natural LDL (N-LDL) and MG-glycatedLDL to probe its immunogenicity. The glycoxidative lesion detection in LDL of DM with disease duration (D.D.) of 5-15 years and D.D. > 15 years was found to be significantly higher as compared to normal healthy subjects (NHS) LDL. The findings support the notion that prolonged duration of diabetes can cause structural alteration in LDL protein molecules, rendering them highly immunogenic in nature. The presence of LDL lesions specific to MG-associated glycoxidation would further help in assessing the progression of diabetes mellitus.

Regulation of low-density lipoprotein on lipid metabolism in macrophages of large yellow croaker (Larimichthys crocea)

Low-density lipoprotein (LDL) is the main carrier of cholesterol transport in plasma, which participates in regulating lipid homeostasis. Studies in mammals have shown that high levels of LDL in plasma absorbed by macrophages trigger the formation of lipid-rich foam cells, leading to the development of atherosclerotic plaques. Although lipid-rich atherosclerosis-like lesions have been discovered in the aorta of several fish species, the physiological function of LDL in fish macrophages remains poorly understood. In the present study, LDL was isolated from the plasma of large yellow croaker (Larimichthys crocea), and mass spectrometry analysis identified two truncated forms of apolipoprotein B100 in the LDL protein profile. Transcriptomic analysis of LDL-stimulated macrophages revealed that differentially expressed genes (DEGs) were enriched in various pathways related to lipid metabolism, as confirmed by the fact that LDL increased total cholesterol and cholesteryl esters content. Meanwhile, the gene and protein expression levels of perilipin2 (PLIN2), a DEG enriched in the PPAR signaling pathway, were upregulated in response to LDL stimulation. Importantly, knocking down plin2 significantly attenuates LDL-induced cholesterol accumulation and promotes cholesterol efflux. Furthermore, the transcription factor PPARγ, which is upregulated in response to LDL stimulation, can enhance the promoter activity of plin2. In conclusion, this study suggests that LDL may upregulate plin2 expression through PPARγ, resulting in cholesterol accumulation in fish macrophages. This study will facilitate the investigation of the function of LDL in regulating lipid homeostasis in macrophages and shed light on the evolutionary origin of LDL metabolism in vertebrates.

Risk Factors for Left Ventricular Hypertrophy in Patients with Diabetic Kidney Disease: A Multi-Center Study.

One of the earliest echocardiographic features of the left ventricle explored extensivelywas left ventricular hypertrophy (LVH). Numerous studies have identified a few risk factors for LVH, however, there are few for people with diabetic kidney disease (DKD). Therefore, we evaluated the risk factors in DKD patients with LVH by analyzing laboratory data and clinical traits. In total, 500 DKD patients in the Baoding area from February 2016 and June 2020 were admitted and classified as an experimental group (240 cases, LVH group) and a control group (260 cases, non-LVH group). The clinical parameters and laboratory tests of the participants were collected and analyzed retrospectively. Compared with the control group, low-density lipoprotein (LDL), body mass index (BMI), intact parathyroid hormone (iPTH), systolic blood pressure, and 24-hour urine protein were higher in the experimental group (all P<0.01). Multivariable logistic regression analysis results confirmed that high BMI (OR=1.332, 95% CI 1.016-1.537, P=0.006), LDL (OR=1.279, 95% CI 1.008-1.369, P=0.014) and 24-hour urine proteins (OR=1.446, 95% CI 1.104-1.643, P=0.016) were statistically significant. The ROC analysis illustrated that the optimum cutoff value of BMI, LDL, and 24-hour urine proteins for diagnosis of LVH in patients with DKD was 27.36 kg/m2, 4.18 mmol/L, and 1.42 g respectively. The increase in BMI, LDL, and 24-hour urine proteins quantification are independent risk factors for LVH in patients with DKD.

Anti Diabetic Activity of Methanolic and Chloroform Extracts of Solanum seaforthianum In Alloxan Induced Diabetes Rats

Diabetes mellitus is a syndrome characterized by elevated glucose levels, polyurea, polydipsia and weakness due to insufficient entry of available glucose into the cells of the body. There are many allopathic drugs available to manage diabetes such as Sulfonylureas, biguanides, gliptins etc., but associated with hypoglycemia, sweating, dizziness, confusion, hunger, weight gain on long term usage and tolerance will be developed which are further treated using multiple drugs. According to our ancient system of medicine there are many medicinal plants available to treat diabetes with the same potency of allopathic drugs. Natural products are considered to be an effective alternative for the treatment and management of diabetes, because of higher efficacy which initiated the present study of anti diabetic activity of Solanum seaforthianum in an alloxan induced diabetes male albino rat. Diabetes was successfully induced with the help of alloxan at a dose of 120 mg/kg body weight and the animals which are possessing the glucose levels more than 250 mg/dL included in the study. Diabetic induced rats were treated with methanolic and chloroform extracts (200 and 400 mg/kg/kg body weight) of Solanum seaforthianum for 10 days. The collected blood samples on 1st, 4th, 7th, and 10th day were used to determine serum biochemical parameters such as cholesterol, triglycerides, low-density lipoprotein (LDL), high-density lipoprotein (HDL) cholesterol, urea, creatinine and total protein. Results: The treatment with extracts of Solanum seaforthianum significantly reduced the elevated blood glucose levels and reversed other altered parameters of Cholesterol, Triglycerides, HDL, LDL, Urea, Creatinine and Total Protein. Among the two extracts of Solanum seaforthianum chloroform extract has shown comparatively significant (p&lt;0.001) anti diabetic activity in alloxan induced diabetes in albino rats.&#x0D; , , , ,

PENAMBATAN MOLEKULER SENYAWA CENDAWAN ENDOFIT Trichoderma sp. SEBAGAI INHIBITOR PROTEIN LOW DENSITY LIPOPROTEIN, ENZIM LANASTEROL 14 DEMETILASE DAN LIPASE YANG BERTANGGUNG JAWAB DALAM DERMATITIS SEBOROIK

Seborrheic dermatitis (SD) occurs due to excessive sebum production resulting in the growth of the fungus Mellasezia sp. on the skin becomes uncontrollable. Some of the receptors that play a role in SD regulation are Low Density Lipoprotein (LDL) protein, Lanasterol 14-α Demetylase enzyme, and Lipase enzyme. This study aims to determine the potential compounds of the endophytic fungi Trichoderma sp. as an antiseborrheic and knowing the interaction between ligands as inhibitors in SD regulation. The research was conducted with computer-based molecular docking. The results showed that there were two inhibitor candidate compounds based on the selection by binding affinity value, namely Chloromycetin and Benzyl benzoate. Based on the results of docking with VINA autodock, the potential compounds were Benzyl benzoate on LDL protein with binding affinity values of -7.3 kcal/mol, compounds were Chloromycethin on Lipase enzyme with binding affinity values of -7.3 kcal/mol and Lanasterol 14-α demethylase enzyme with a binding affinity value of -7.6 kcal/mol. These compounds form hydrogen bonds and hydrophobic bonds. Based on the prediction of Lipinski's rule of five, the two compounds are effective orally used as anti seborrheic dermatitis drugs

Heterogeneity of T Cells in Atherosclerosis Defined by Single-Cell RNA-Sequencing and Cytometry by Time of Flight.

The infiltration and accumulation of pro- and anti-inflammatory leukocytes within the intimal layer of the arterial wall is a hallmark of developing and progressing atherosclerosis. While traditionally perceived as macrophage- and foam cell-dominated disease, it is now established that atherosclerosis is a partial autoimmune disease that involves the recognition of peptides from ApoB (apolipoprotein B), the core protein of LDL (low-density lipoprotein) cholesterol particles, by CD4+ T-helper cells and autoantibodies against LDL and ApoB. Autoimmunity in the atherosclerotic plaque has long been understood as a pathogenic T-helper type-1 driven response with proinflammatory cytokine secretion. Recent developments in high-parametric cell immunophenotyping by mass cytometry, single-cell RNA-sequencing, and in tools exploring antigen-specificity have established the existence of several unforeseen layers of T-cell diversity with mixed TH1 and T regulatory cells transcriptional programs and unpredicted fates. These findings suggest that pathogenic ApoB-reactive T cells evolve from atheroprotective and immunosuppressive CD4+ T regulatory cells that lose their protective properties over time. Here, we discuss T-cell heterogeneity in atherosclerosis with a focus on plasticity, antigen-specificity, exhaustion, maturation, tissue residency, and its potential use in clinical prediction.

Dose and duration-dependent toxicological evaluation of lead acetate in chicks.

Lead is one of the utmost contaminated and dangerous heavy metals. This toxicant ultimately enters into the human body through the food chain and accumulated in the body because the animal/human body has not an appropriate mechanism to excrete it from the body. The main objective of the present research was to assess the toxicological effects of lead on body weights, biochemical, and hematological parameters of chickens and also to measure its bioaccumulation in the brain. Lead acetate was administrated orally at doses of 0, 71, 142, 213, and 284mg/kg of body weight of chicken for groups A, B, C, D, and E, respectively. Along with determination of biometry of all experimental chicks, hematological [hemoglobin (Hb), packed cell volume (PCV), mean corpuscular hemoglobin concentration (MCHC), total erythrocyte count (TEC), white blood cells (WBCs), leukocyte differential count (LDC)] and biochemical [low density lipoprotein (LDL), total protein, high-density lipoprotein (HDL), and alanine aminotransferase (ALT)] parameters were measured. The present study showed that the bodyweight of chickens was not affected significantly by lead acetate exposure. The levels of MCHC, PCV, TEC, Hb, LDL, HDL, and total protein were found to be significantly decreased while WBC, LDC, and ALT profile were enhanced due to administration of lead acetate. Bioaccumulation of lead acetate was found to be higher in the brain. We conclude that the chronic administration of lead acetate affected the blood and biochemical profile of exposed chicken. These effects might be due to the accumulation of the chemical in certain vital organ(s). However, further studies in the future are suggested to refine such findings.

Ambient Air Pollution Is Associated With HDL (High-Density Lipoprotein) Dysfunction in Healthy Adults.

Objective- We aimed to assess whether exposure to higher levels of ambient air pollution impairs HDL (high-density lipoprotein) function and to elucidate the underlying biological mechanisms potentially involved. Approach and Results- In the Beijing AIRCHD study (Air Pollution and Cardiovascular Dysfunction in Healthy Adults), 73 healthy adults (23.3±5.4 years) were followed-up with 4 repeated study visits in 2014 to 2016. During each visit, ambient air pollution concentrations, HDL function metrics, and parameters of inflammation and oxidative stress were measured. Average daily concentrations of ambient particulate matter in diameter <2.5 μm were 62.9 µg/m3 (8.1-331.0 µg/m3). We observed significant decreases in HDL cholesterol efflux capacity of 2.3% (95% CI, -4.3 to -0.3) to 5.0% (95% CI, -7.6 to -2.4) associated with interquartile range increases in moving average concentrations of particulate matter in diameter <2.5 μm and traffic-related air pollutants (black carbon, nitrogen dioxide, and carbon monoxide) during the 1 to 7 days before each participant's clinic visit. Higher ambient air pollutant levels were also associated with significant reductions in circulating HDL cholesterol and apoA-I (apolipoprotein A-I), as well as elevations in HDL oxidation index, oxidized LDL (low-density lipoprotein), malondialdehyde, and high-sensitivity C-reactive protein. Conclusions- Higher ambient air pollution concentrations were associated with impairments in HDL functionality, potentially because of systemic inflammation and oxidative stress. These novel findings further our understanding of the mechanisms whereby air pollutants promote cardiometabolic disorders.

Low-Density Lipoprotein-Reactive T Cells Regulate Plasma Cholesterol Levels and Development of Atherosclerosis in Humanized Hypercholesterolemic Mice

Atherosclerotic cardiovascular disease is a chronic inflammatory process initiated when cholesterol-carrying low-density lipoprotein (LDL) is retained in the arterial wall. CD4+ T cells, some of which recognize peptide components of LDL as antigen, are recruited to the forming lesion, resulting in T-cell activation. Although these T cells are thought to be proatherogenic, LDL immunization reduces disease in experimental animals. These seemingly contradictory findings have hampered the development of immune-based cardiovascular therapy. The present study was designed to clarify how activation of LDL-reactive T cells impacts on metabolism and vascular pathobiology. We have developed a T-cell receptor-transgenic mouse model to characterize the effects of immune reactions against LDL. Through adoptive cell transfers and cross-breeding to hypercholesterolemic mice expressing the antigenic human LDL protein apolipoprotein B-100, we evaluate the effects on atherosclerosis. A subpopulation of LDL-reactive T cells survived clonal selection in the thymus, developed into T follicular helper cells in lymphoid tissues on antigen recognition, and promoted B-cell activation. This led to production of anti-LDL immunoglobulin G antibodies that enhanced LDL clearance through immune complex formation. Furthermore, the cellular immune response to LDL was associated with increased cholesterol excretion in feces and with reduced vascular inflammation. These data show that anti-LDL immunoreactivity evokes 3 atheroprotective mechanisms: antibody-dependent LDL clearance, increased cholesterol excretion, and reduced vascular inflammation.

Regulatory CD4+ T Cells Recognize Major Histocompatibility Complex Class II Molecule-Restricted Peptide Epitopes of Apolipoprotein B.

CD4+ T cells play an important role in atherosclerosis, but their antigen specificity is poorly understood. Immunization with apolipoprotein B (ApoB, core protein of low density lipoprotein) is known to be atheroprotective in animal models. Here, we report on a human APOB peptide, p18, that is sequence-identical in mouse ApoB and binds to both mouse and human major histocompatibility complex class II molecules. We constructed p18 tetramers to detect human and mouse APOB-specific T cells and assayed their phenotype by flow cytometry including CD4 lineage transcription factors, intracellular cytokines, and T cell receptor activation. Apolipoprotein E-deficient ( Apoe-/-) mice were vaccinated with p18 peptide or adjuvants alone, and atherosclerotic burden in the aorta was determined. In human peripheral blood mononuclear cells from donors without cardiovascular disease, p18 specific CD4+ T cells detected by a new human leukocyte antigen-antigen D related-p18 tetramers were mostly Foxp3+ regulatory T cells (Tregs). Donors with subclinical cardiovascular disease as detected by carotid artery ultrasound had Tregs coexpressing retinoic acid-related orphan receptor gamma t or T-bet, which were both almost absent in donors without cardiovascular disease. In Apoe-/- mice, immunization with p18 induced Tregs and reduced atherosclerotic lesions. After peptide restimulation, responding CD4+ T cells identified by Nur77-GFP (green fluorescent protein) were highly enriched in Tregs. A new mouse I-Ab-p18 tetramer identified the expansion of p18-specific CD4+ T cells on vaccination, which were enriched for interleukin-10-producing Tregs. These findings show that APOB p18-specific CD4+ T cells are mainly Tregs in healthy donors, but coexpress other CD4 lineage transcription factors in donors with subclinical cardiovascular disease. This study identifies ApoB peptide 18 as the first Treg epitope in human and mouse atherosclerosis.

MHC class II tetramers identify apolipoprotein B-specific CD4 T cells in mice and humans

Abstract Immunization with atherosclerotic plaque antigens including MHC-II-restricted peptides from apolipoprotein B (ApoB, core protein of low density lipoprotein, LDL) is known to be atheroprotective in animal models. Herein we report a human ApoB peptide whose sequence is identical to mouse ApoB and which binds to both mouse (I-Ab) and human MHC class II. Immunization with this peptide reduced atherosclerotic plaque in the aortas of Apoe−/− mice fed with a Western diet and induced IL-10 producing CD25+FoxP3+ regulatory T cells (Tregs). After peptide restimulation in vivo, responding CD4+ T cells (by Nur77-GFP) were highly enriched in CD25+FoxP3+ Tregs. Using a novel I-Ab tetramer reagent, we further showed that peptide-specific CD4+ T cells expand upon vaccination, ~50% express FoxP3, and were 10-fold enriched for IL-10 producers. In human peripheral blood mononuclear cells (PBMCs) from individuals assessed for subclinical cardiovascular disease, ApoB-specific CD4+ T cells were detected using novel HLA-DR1/ApoB peptide tetramers. Peptide-specific CD4+ T cells from donors with carotid artery plaques showed more RORγt expression and less FoxP3 expression compared to those without. This is the first demonstration of ApoB-specific CD4+ T cells by tetramer in humans and the first animal model evidence of atheroprotective vaccination with a human ApoB peptide.

Atheroprotective vaccination with MHC-II-restricted ApoB peptides induces peritoneal IL-10-producing CD4 T cells.

Although immunization with major histocompatibility complex (MHC) class II-restricted apolipoprotein B (ApoB) peptides has been shown to be atheroprotective, the mechanism is unclear. Here, we investigated CD4+ T cell populations in immunized atherosclerotic mice. Peptides (16-mers) from mouse ApoB, the core protein of low-density lipoprotein (LDL), were screened for binding to I-Ab by computer prediction and confirmed by radiolabeled peptide competition. Three new peptides, P101 (FGKQGFFPDSVNKALY, 5.5 nM IC50), P102 (TLYALSHAVNSYFDVD, 6.8 nM), and P103 (LYYKEDKTSLSASAAS, 95 nM), were tested in an atherosclerosis model (Apoe-/- mice on Western diet). Immunization with each of the three peptides (1 time in complete Freund's adjuvant subcuntaneously and 4 time in incomplete Freund's adjuvant intraperitoneally) but not with adjuvant alone showed significantly reduced atherosclerotic plaques in the aortic root by serial sections and in the whole aorta by en face staining. There were no differences in body weight, LDL cholesterol, or triglycerides. Peritoneal leukocytes from ApoB peptide-immunized mice, but not control mice, secreted significant amounts of IL-10 (150 pg/ml). Flow cytometry showed that peptide immunization induced IL-10 in 10% of peritoneal CD4+ T cells, some of which also expressed chemokine (C-C motif) receptor 5 (CCR5). Vaccination with ApoB peptides expanded peritoneal FoxP3+ regulatory CD4+ T cells and more than tripled the number of CCR5+FoxP3+ cells. Similar trends were also seen in the draining mediastinal lymph nodes but not in the nondraining inguinal lymph nodes. We conclude that vaccination with MHC class II-restricted autologous ApoB peptides induces regulatory T cells (Tregs) and IL-10, suggesting a plausible mechanism for atheroprotection.NEW & NOTEWORTHY Vaccination against apolipoprotein B (ApoB), the protein of LDL, attracts attention as a novel approach to prevent atherosclerosis. We discovered major histocompatibility complex class II-restricted ApoB peptides, which reduce atherosclerosis and induce IL-10-producing CD4+ T cells and chemokine (C-C motif) receptor 5 expression on regulatory T cells, suggesting that immunization with ApoB peptides inhibits atherosclerosis by inducing anti-inflammatory cytokines.

My Early Days with Ari Helenius: Detergents and Viruses.

My Early Days with Ari Helenius: Detergents and Viruses.

Differential urinary glycoproteome analysis of type 2 diabetic nephropathy using 2D-LC-MS/MS and iTRAQ quantification.

BackgroundDiabetic nephropathy (DN) is the leading cause of chronic kidney failure and end-stage kidney disease. More accurate and non-invasive test for the diagnosis and monitoring the progression of DN is urgently needed for the better care of such patients.MethodsIn this study we utilized urinary glycoproteome to discover the differential proteins during the course of type 2 DN. The urinary glycoproteins from normal controls, normalbuminuira, microalbuminura, and macroalbuminuria patients were enriched by concanavalin A (ConA) and analyzed by 2DLC/MS/MS and isobaric tags for relative and absolute quantitation quantification.ResultsA total of 478 proteins were identified and 408 were annotated as N-linked glycoproteins. A total of 72, 107 and 123 differential proteins were identified in normalbuminuria, microalbuminuria and macroalbuminuria, respectively. By bioinformatics analysis, in normalbuminruia state, cell proliferation and cell movement were activated, which might reflect the compensatory phase during the disease development. In micro- and macro-albuminuria, cell death and apoptosis was activated, which might reflect the de-compensatory phase. Pathway analysis showed acute phase proteins, the member of high density lipoprotein and low density lipoprotein proteins were changed, indicating the role of the inflammatory response and lipid metabolism abnormality in the pathogenesis of DN. Six selected differential proteins were validated by Western Blot. Alpha-1-antitrypsin (SERPINA1) and Ceruloplasmin are the two markers with excellent area under curve values (0.929 and 1.000 respectively) to distinguish the microalbuminuria and normalbuminuria. For the first time, we found pro-epidermal growth factor and prolactin-inducible protein were decreased in macroalbuminuria stage, which might reflect the inhibition of cell viability and the activation of cell death in kidney.ConclusionsAbove data indicated that urinary glycoproteome could be useful to distinguish the differences in protein profiles in different stages in DN, which will help better individualized care of patients in DN.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-015-0712-9) contains supplementary material, which is available to authorized users.

α-Tocopherol bioavailability is lower in adults with metabolic syndrome regardless of dairy fat co-ingestion: a randomized, double-blind, crossover trial

Increasing dietary fat intake is expected to improve α-tocopherol bioavailability, which could be beneficial for improving α-tocopherol status, especially in cohorts at high cardiometabolic risk who fail to meet dietary α-tocopherol requirements. Our objective was to assess dose-dependent effects of dairy fat and metabolic syndrome (MetS) health status on α-tocopherol pharmacokinetics in plasma and lipoproteins. A randomized, crossover, double-blind study was conducted in healthy and MetS adults (n = 10/group) who ingested encapsulated hexadeuterium-labeled (d6)-RRR-α-tocopherol (15 mg) with 240 mL nonfat (0.2 g fat), reduced-fat (4.8 g fat), or whole (7.9 g fat) milk before blood collection at regular intervals for 72 h. Compared with healthy participants, those with MetS had lower (P < 0.05) baseline plasma α-tocopherol (μmol/mmol lipid) and greater oxidized low-density lipoprotein (LDL), interleukin (IL)-6, IL-10, and C-reactive protein. Regardless of health status, d6-α-tocopherol bioavailability was unaffected by increasing amounts of dairy fat provided by milk beverages, but MetS participants had lower estimated d6-α-tocopherol absorption (±SEM) than did healthy participants (26.1% ± 1.0% compared with 29.5% ± 1.1%). They also had lower plasma d6-α-tocopherol AUC from 0 to 72 h, as well as maximal concentrations (Cmax: 2.04 ± 0.14 compared with 2.73 ± 0.18 μmol/L) and slower rates of plasma disappearance but similar times to Cmax. MetS participants had lower d6-α-tocopherol AUC from t = 0-12 h (AUC0- t final) in lipoprotein fractions [chylomicron, very-low-density lipoprotein (VLDL), LDL, high-density lipoprotein]. Percentages of d6-α-tocopherol AUC0- t final in both the chylomicron (r = -0.46 to -0.52) and VLDL (r = -0.49 to -0.68) fractions were inversely correlated with oxidized LDL, IL-10, IL-6, and C-reactive protein. At dietary intakes equivalent to the Recommended Dietary Allowance, α-tocopherol bioavailability is unaffected by dairy fat quantity but is lower in MetS adults, potentially because of greater inflammation and oxidative stress that limits small intestinal α-tocopherol absorption and/or impairs hepatic α-tocopherol trafficking. These findings support higher dietary α-tocopherol requirements for MetS adults. This trial was registered at www.clinicaltrials.gov as NCT01787591.

Analysis of the expression of HMGB-1, CXCL16, miRNA-30a, and TGF-β1 in primary nephritic syndrome patients and its significance.

We investigated the expression levels of high-mobility group box protein 1 (HMGB-1), CXC chemokine ligand 16 (CXCL16), microRNA (miRNA)-30a and transforming growth factor ß1 (TGF-ß1) in primary nephritic syndrome (PNS) patients and the clinical significance of this expression. A total of 56 patients with PNS were included in the PNS group, while 50 healthy subjects formed the normal control group. Serum levels of HMGB-1, CXCL16, miRNA-30a, and urinary TGF-ß1 concentrations were quantified along with other biochemical indices, including serum albumin, triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein, and urinary proteins. The correlation between levels of HMGB-1, CXCL16, miRNA-30a, and TGF-ß1 and biochemical indexes was further analyzed. PNS group patients had significantly higher levels of HMGB-1, CXCL16, miRNA- 30a, and TGF-ß1 compared to the control group (P < 0.05). PNS patients also had higher 24-h urinary protein, TG, TC, and LDL levels but lower serum albumin compared to subjects in the control group (P < 0.05). Serum HMGB-1, CXCL16, miRNA-30a, and urinary TGF-ß1 levels were all negatively correlated with serum albumin levels, but were positively correlated with TG, TC, LDL, and 24-h urinary protein (P < 0.05 in all cases). Additionally, a positive correlation existed among serum HMGB-1, CXCL16, miRNA-30a, and urinary TGF-ß1 levels (P < 0.01). HMGB-1, CXCL16, miRNA-30a, and urinary TGF-ß1 were highly expressed in PNS patients and may play important roles in the pathogenesis and development of PNS.

Naturally Occurring Autoantibodies to Oxidation-Specific Epitopes in Patients with Venous Thromboembolism

Background: Naturally occurring IgM autoantibodies (NAb) recognize oxidation-specific epitopes which are altered self structures on apoptotic cells, oxidized LDL, or vessel wall proteins. Oxidation-specific epitopes are proinflammatory antigens and potential markers for chronic inflammatory conditions. The risk of venous thromboembolism (VTE) is associated with (pro)inflammatory states such as dyslipoproteinemia, obesity or cancer. We hypothesize that natural antibodies may target epitopes which are also involved in the pathogenesis of VTE. In pursuing a “proof of concept”, we studied Nab in patients with a high venous thrombotic risk and evaluated their association with the risk of recurrent VTE.Methods: We determined Nab levels in 663 patients with a first unprovoked deep vein thrombosis and/or pulmonary embolism who were prospectively followed after discontinuation of anticoagulation in the Austrian Study on Recurrent Venous Thromboembolism, a multicentre cohort study. We excluded those with coagulation inhibitor deficiencies, lupus anticoagulant, cancer, pregnancy, long-term antithrombotic therapy, or homozygosity/double heterozygosity for factor V Leiden and/or the prothrombin mutation. Study end point was recurrent VTE.Nab target different oxidation-specific epitopes including phosphorylcholine-containing oxidized phospholipids which were generated by oxidation of low density lipoproteins (LDL) with CuSO4. Levels of IgM against oxidized (Ox) LDL were determined in venous blood drawn three weeks after discontinuation of anticoagulants by chemiluminescent enzyme linked immunoabsorbent assay and expressed as relative light units (RLU)/100ms.To determine the effect of anti-OxLDL IgM on the recurrence risk, models for time-to-event-data were used. Their direct effect on the recurrence risk was estimated by a cause-specific Cox proportional-hazards model, followed by a Fine-Gray model to account for competing events (death, restart of antithrombotics) and expressed by subdistribution hazard ratios (SHR). Anti-OxLDL IgM levels are given as median (25th, 75th percentiles).Results: Levels of anti-OxLDL IgM in patients with a first unprovoked VTE were 20.17 RLU/100ms (12.26, 34.72). Levels did not correlate with either age (rs=-0.16) or BMI (rs=-0.16). Levels correlated with sex [higher in women; 24.24 (14.6, 40.0) vs. 16.76 (10.46, 27.9) RLU/100ms, p<0.001]. 151 (22.8%) of 663 patients (mean age 48 years, 54% women) had recurrence during a median of 67 months. Anti-OxLDL IgM levels were higher in patients without recurrence [20.84 (12.32, 36.85) vs. 17.60 (11.97, 26.93) RLU/100ms, p=0.02]. For each doubling of anti-OxLDL IgM levels, the SHR of recurrence was 0.85 (95% CI 0.74-0.98; p = 0.02) and was 0.95 (0.82-1.1; p=0.5) after adjustment for sex.Patients with anti-OxLDL IgM levels within the second tercile (14.6 - <28.64 RLU/100ms) had a similar recurrence risk as patients with levels within the first tercile (<14.60 RLU/100ms) (SHR 0.93, 95% CI 0.65–1.33, p=0.7). The recurrence risk was significantly lower in patients with anti-OxLDL IgM levels within the third tercile (>28.64 RLU/100ms) (SHR 0.58, 95% CI 0.38–0.87, p<0.01). After adjustment for sex, the recurrence risk was still lower among patients with high anti-OxLDL IgM (SHR 0.75, 95% CI 0.49–1.14, p=0.2) but without statistical significance.We stratified patients according to an anti-OxLDL IgM level corresponding to the 60th percentile (14.6 RLU/100ms). The cumulative probability of recurrence after 5 years in patients with anti-OxLDL IGM > 60th percentile was 12.3% (8.5-16.9%) and was 20.3% (16.4-24.6%) among those with lower levels (p<0.001) (Figure). The corresponding SHR was 0.55 (0.38-0.78; p <0.001) for higher anti-OxLDL IgM and was 0.67 (95% CI 0.47-0.96; p=0.03) after adjustment for sex.Conclusions: An association exists between anti-OxLDL IgM and VTE recurrence which is consistent with a protective function of NAb via their ability to bind proinflammatory oxidation-specific epitopes. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.