Association of lipid parameters with insulin resistance in the Kazakh population.

BACKGROUND: Insulin resistance (IR) is a consequence of chronic adipose tissue inflammation and underlies the pathogenesis of several diseases, such as type 2 diabetes mellitus, cardiovascular diseases and metabolic syndrome. In this study, we examined the association between dyslipidaemia and IR; directly comparing conventional lipid ratios and apoB/apoA1 ratios for strength and independence as risk factors for IR in a Kazakh population. METHODS: The design of this study was a case-control study. There were 507 participants in the study. We examined each participant's plasma total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, apolipoprotein B, and apolipoprotein A1. IR was determined using an IR homeostasis model assessment (HOMA-IR). To assess the risk of an atherogenic blood lipid profile, atherogenicity coefficients were calculated: Bad cholesterol to good cholesterol ratio ((TC-HDL)/HDL); TG to HDL ratio (TRG/HDL); apoB to apoA1 ratio (apoB/apoA1). RESULTS: In this study, high waist circumference and BMI were more common in men. The group with IR had significantly higher waist circumference (cm) (p = 0.0001) and BMI (kg/m2) (p = 0.04) than the group without IR. The risk of IR was significantly associated with the apoB/apoA1 ratio (p = 0.03). Analysis of the association between HOMA-IR and apoB/apoA1 ratio increased the risk of IR at apoB/apoA1 ratios of 0.71 to 0.85 and above 0.86 by a factor of 1.93 and 1.84, respectively. HOMA-IR levels were weakly significantly correlated with TG levels (rS = 0.3; p = 0.0001) and very weakly positively correlated with apoB levels (rS = 0.1; p = 0.002) and apoB/apoA1 (rS = 0.1; p = 0.001), there was a weak negative correlation with apoA1 levels (rS = -0.1; p = 0.02). Logistic regression analysis showed that the risk of developing IR was significantly lower in men than in women, adjusted OR (95% CI) = 0.75 (0.49-1.0) p = 0.02. CONCLUSION: In our study, IR was more common in Kazakh women than in Kazakh men. IR was also associated with apoB and TG levels. Thus, we suggest that analysis of TG, apoB and apoB/apoA1 ratio may be recommended as early predictors of IR risk in the Kazakh population (Tab. 3, Ref. 22). Text in PDF www.elis.sk Keywords: insulin resistance, dyslipidaemia, apolipoproteins, triglycerides, lipids.

Hair mercury is associated with dyslipidemia and cardiovascular risk: An anthropometric, biochemical and genetic cross-sectional study of Amazonian vulnerable populations.

This cross-sectional study evaluated the association between human exposure to mercury and cardiovascular risk using lipid profile (including apolipoproteins) and genetic analysis of Amazonian riverine population. Anthropometric data (gender, age, height, weight, blood pressure, and neck and waist circumferences) of the participants were recorded. Total mercury and methylmercury (MeHg) content were quantified in hair by ICP-MS and GC-pyro-AFS system. Polymorphisms rs662799, rs693, rs429358 and rs7412 (of genes of apolipoproteins A-V, B, and E at positions 112 and 158, respectively) were genotyped by real-time PCR. The population presented a dyslipidemia profile significantly correlated with high mercury levels. The apolipoprotein B/apolipoprotein A-I (ApoB/ApoA-I) index was also positively correlated with mercury, supporting a possible causal relationship. Allelic distributions were similar to those described in other populations, suggesting that genetic susceptibility may not have a significant role in the lipid alterations found in this work. This study demonstrated for the first time: i) the relationship between mercury exposure and cardiovascular risk-related apolipoproteins in humans, ii) the ApoB levels and the ApoB/ApoA-I index as the risk factors more strongly associated to the mercury-related dyslipidemia in humans, and iii) the prevalence of high/moderate risk of acute myocardial infarction in the vulnerable and chronically exposed-populations of the Amazon, in addition to the genotypic profile of the three most frequent polymorphisms in apolipoproteins of relevance for cardiovascular risk. This early detection of lipid alterations is essential to prevent the development of cardiovascular diseases (CVD), especially in chronically exposed populations such as those found in the Amazon. Therefore, in addition to provide data for the Minamata Convention implementation, our work is in line with the efforts joined by all members of the World Health Organization committed to reducing premature deaths originating from non-communicable diseases by 25% in 2025, including CVD.

Cargo Receptor-Mediated ER Export in Lipoprotein Secretion and Lipid Homeostasis.

APOB-containing lipoproteins are large, complex lipid carriers that ferry bulk lipids into the circulation via the secretory pathway, originating from the endoplasmic reticulum of specialized cells in the liver or the gut. Elevation of APOB-containing lipoproteins in the plasma represents a major risk factor for cardiovascular diseases. The production of these lipoproteins requires enzyme-catalyzed, cross-membrane transfer of neutral lipids and phospholipids to lipoproteins, in particular onto the structural component APOB. Transport of these lipid-bearing cargos relies on the COPII machinery and employs the transmembrane cargo receptor SURF4 and the small GTPase SAR1B, together constituting a selective transport program. Intriguingly, a number of factors implicated in lipoprotein production are also packaged into COPII vesicles and may be cotransported with APOB. These observations therefore point to a specialized produce-and-export itinerary during the secretion of these lipid-bearing cargos, warranting future investigations into this unique yet pivotal process at the crossroad of cell biology and physiology.

ApoA-I and ApoB levels, and ApoB-to-ApoA-I ratio as candidate pre-treatment biomarkers of pathomorphological response to neoadjuvant therapy in gastric and esophago-gastric junction adenocarcinoma.

<b><br>Introduction:</b> Neoadjuvant chemotherapy (NAC) is a part of the current standard of care in a locally advanced gastric adenocarcinoma (GA) and esophagogastric junction adenocarcinoma (EGJA), but only patients with good pathomorphological response (pR) to NAC benefit from prolonged overall survival.</br> <b><br>Aim:</b> The study aims to evaluate ApoA-I and ApoB as candidate pre-treatment biomarkers of pR to NAC in patients with GA and EGJA.</br> <b><br>Materials and methods:</b> Serum samples were collected from 18 patients with GA and 9 with EGJA before the initiation of NAC to determine the ApoA-I and ApoB levels. After NAC tumor regression grade (TRG) was evaluated in resected specimens according to the Mandard's tumor regression grading system and correlated with pre-treatment ApoA-I and ApoB serum concentration, and ApoB-to-ApoA-I serum concentration ratio.</br> <b><br>Results:</b> We found a positive correlation of ApoA-I level and pR (95% CI: -0.863 to -0.467; P < 0.0001), a negative correlation of ApoB level and pR (95% CI: 0.445 to 0.857; P < 0.0001), a negative correlation of ApoB-to-ApoA-I ratio and pR (95% CI: 0.835 to 0.964; P < 0.0001).</br> <b><br>Conclusions:</b> ApoA-I and ApoB levels, and ApoB-to-ApoA-I ratio are candidate pre-treatment predictors of pR to NAC in GA and may help to guide personalized therapy.</br>Our work fits into the dynamically developing trend of personalized treatment. It describes a potentially important rationale for further evaluation of apolipoprotein A-I and apolipoprotein B as predictors of cancer response to neoadjuvant therapy.

Associations of Fine Particulate Matter Constituents with Metabolic Syndrome and the Mediating Role of Apolipoprotein B: A Multicenter Study in Middle-Aged and Elderly Chinese Adults.

Fine particulate matter (PM2.5) was reported to be associated with metabolic syndrome (MetS), but how PM2.5 constituents affect MetS and the underlying mediators remains unclear. We aimed to investigate the associations of long-term exposure to 24 kinds of PM2.5 constituents with MetS (defined by five indicators) in middle-aged and elderly adults and to further explore the potential mediating role of apolipoprotein B (ApoB). A multicenter study was conducted by recruiting subjects (n = 2045) in the Beijing-Tianjin-Hebei region from the cohort of Sub-Clinical Outcomes of Polluted Air in China (SCOPA-China Cohort). Relationships among PM2.5 constituents, serum ApoB levels, and MetS were estimated by multiple logistic/linear regression models. Mediation analysis quantified the role of ApoB in "PM2.5 constituents-MetS" associations. Results indicated PM2.5 was significantly related to elevated MetS prevalence. The MetS odds increased after exposure to sulfate (SO42-), calcium ion (Ca2+), magnesium ion (Mg2+), Si, Zn, Ca, Mn, Ba, Cu, As, Cr, Ni, or Se (odds ratios ranged from 1.103 to 3.025 per interquartile range increase in each constituent). PM2.5 and some constituents (SO42-, Ca2+, Mg2+, Ca, and As) were positively related to serum ApoB levels. ApoB mediated 22.10% of the association between PM2.5 and MetS. Besides, ApoB mediated 24.59%, 50.17%, 12.70%, and 9.63% of the associations of SO42-, Ca2+, Ca, and As with MetS, respectively. Our findings suggest that ApoB partially mediates relationships between PM2.5 constituents and MetS risk in China.

Integrated weighted gene co-expression network analysis reveals biomarkers associated with prognosis of high-grade serous ovarian cancer.

BACKGROUND: Ovarian cancer is the gynecologic tumor with the highest fatality rate, and high-grade serous ovarian cancer (HGSOC) is the most common and malignant type of ovarian cancer. One important reason for the poor prognosis of HGSOC is the lack of effective diagnostic and prognostic biomarkers. New biomarkers are necessary for the improvement of treatment strategies and to ensure appropriate healthcare decisions. METHODS: To construct the co-expression network of HGSOC samples, we applied weighted gene co-expression network analysis (WGCNA) to assess the proteomic data obtained from the Clinical Proteomic Tumor Analysis Consortium (CPTAC), and module-trait relationship was then analyzed and plotted in a heatmap to choose key module associated with HGSOC. Subsequently, hub genes with high connectivity in key module were identified by Cytoscape software. Furthermore, the biomarkers were selected through survival analysis, followed by evaluation using the relative operating characteristic (ROC) analysis. RESULTS: A total of 9 modules were identified by WGCNA, and module-trait analysis revealed that the brown module was significantly associated with HGSOC (cor = 0.7). Ten hub genes with the highest connectivity were selected by protein-protein interaction analysis. After survival and ROC analysis, ALB, APOB and SERPINA1 were suggested to be the biomarkers, and their protein levels were positively correlated with HGSOC prognosis. CONCLUSION: We conducted the first gene co-expression analysis using proteomic data from HGSOC samples, and found that ALB, APOB and SERPINA1 had prognostic value, which might be applied for the treatment of HGSOC in the future.

A comparative study on the lipidome of normal knee synovial fluid from humans and horses.

The current limitations in evaluating synovial fluid (SF) components in health and disease and between species are due in part to the lack of data on normal SF, because of low availability of SF from healthy articular joints. Our study aimed to quantify species-dependent differences in phospholipid (PL) profiles of normal knee SF obtained from equine and human donors. Knee SF was obtained during autopsy by arthrocentesis from 15 and 13 joint-healthy human and equine donors, respectively. PL species extracted from SF were quantitated by mass spectrometry whereas ELISA determined apolipoprotein (Apo) B-100. Wilcoxon's rank sum test with adjustment of scores for tied values was applied followed by Holm´s method to account for multiple testing. Six lipid classes with 89 PL species were quantified, namely phosphatidylcholine, lysophosphatidylcholine, sphingomyelin, phosphatidylethanolamine, plasmalogen, and ceramide. Importantly, equine SF contains about half of the PL content determined in human SF with some characteristic changes in PL composition. Nutritional habits, decreased apolipoprotein levels and altered enzymatic activities may have caused the observed different PL profiles. Our study provides comprehensive quantitative data on PL species levels in normal human and equine knee SF so that research in joint diseases and articular lubrication can be facilitated.

Dual size-exclusion chromatography for efficient isolation of extracellular vesicles from bone marrow derived human plasma.

Isolation of pure extracellular vesicles (EVs), especially from blood, has been a major challenge in the field of EV research. The presence of lipoproteins and soluble proteins often hinders the isolation of high purity EVs upon utilization of conventional separation methods. To circumvent such problems, we designed a single-step dual size-exclusion chromatography (dSEC) column for effective isolation of highly pure EVs from bone marrow derived human plasma. With an aim to select appropriate column design parameters, we analyzed the physiochemical properties of the major substances in bone marrow derived plasma, which include EVs, lipoproteins, and soluble proteins. Based on these findings, we devised a novel dSEC column with two different types of porous beads sequentially stacked each other for efficient separation of EVs from other contaminants. The newly developed dSEC columns exhibited better performance in isolating highly pure EVs from AML plasma in comparison to conventional isolation methods.

Association of dyslipidaemia with Alzheimer's disease in a cohort of postmenopausal women.

OBJECTIVE: To evaluate the association between dyslipidaemia and Alzheimer's disease (AD) in a cohort of postmenopausal women. METHODS: This retrospective study analysed data from postmenopausal women with early AD (group AD) and a cohort of healthy age- and sex-matched control subjects (group NC) that were considered to be within standard limits according to a neuropsychological assessment between March 2010 and March 2019. The primary endpoints were body mass index and lipid-related laboratory parameters, including leptin, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total cholesterol, adiponectin, triglycerides, apolipoprotein A1, apolipoprotein B and apolipoprotein E4, which were evaluated using multivariate binary logistic analysis. RESULTS: The study enrolled 200 postmenopausal women with early AD (mean ± SD age 69.34 ± 6.25 years) and 180 control subjects (mean ± SD age 67.48 ± 7.42 years). Lower HDL-C and higher LDL-C were risk factors for AD. A multivariate binary logistic regression model demonstrated that lower HDL-C and higher LDL-C were the only variables associated with the development of AD (odds ratio [OR] 21.14, 95% confidence interval [CI] 2.47, 4.13; OR 36.35, 95% CI 1.24, 3.38; respectively). CONCLUSION: Both low HDL-C and high LDL-C were associated with the occurrence of AD in a cohort of postmenopausal women.

Effects of ABCA1 gene polymorphisms on risk factors, susceptibility and severity of coronary artery disease.

BACKGROUND: The relationships between the rs1800976, rs4149313 and rs2230806 polymorphisms in ATP binding cassette protein A1 and severity of coronary artery disease (CAD) remain unclear. METHODS: Four hundred and forty-two patients with CAD and 217 CAD-free subjects were enrolled in this study. The rs1800976, rs4149313 and rs2230806 polymorphisms were genotyped by PCR-RFLP. Severity of CAD was evaluated by Gensini score system, number of stenotic coronary vessels and extent of coronary stenosis. RESULTS: C allele of the rs1800976 polymorphism, G allele of the rs4149313 polymorphism and A allele of the rs2230806 polymorphism were found to be risk alleles for CAD (p<0.05 for all). In patients with CAD, C allele of the rs1800976 polymorphism was associated with high levels of hypersensitive C reactive protein (hs-CRP) and cystatin c (CysC), and its frequency increased with percentiles of Gensini score, number of stenotic coronary vessels and extent of coronary stenosis (p<0.05 for all). The subjects with GA genotype of the rs4149313 polymorphism had higher levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), apolipoprotein B and hs-CRP than those with AA genotype (p<0.05 for all). The subjects with AA genotype of the rs2230806 polymorphism had higher levels of TC, LDL-C and uric acid than those with GA genotype (p<0.05 for all). No associations between the rs4149313 or rs2230806 polymorphism and severity of CAD were detected. CONCLUSIONS: The rs1800976 polymorphism is significantly associated with the occurrence and severity of CAD, which is possibly mediated by hs-CRP and CysC.

Activity of Lipoprotein-Associated Enzymes in Indigenous Children Living at Different Altitudes.

BACKGROUND: High altitude is associated with hypobaric hypoxia, and metabolic modifications. In particular, alterations to lipoprotein-associated enzymes have been reported under hypoxia. OBJECTIVE: To determine the association between paraoxonase 1 (PON-1) and Cholesteryl-ester transfer protein (CETP) activities and altitude in two groups of Argentinean Indigenous schoolchildren living at different altitudes. METHODS: A cross-sectional study compared 151 schoolchildren from San Antonio de los Cobres (SAC), 3,750 m, with 175 schoolchildren from Chicoana (CH), 1,400 m. Anthropometric data, lipids, apolipoprotein (apo) A-I, apo B, plus PON-1 and CETP activities were determined. RESULTS: The prevalence of overweight/obesity was significantly lower in SAC than in CH. Z- BMI (0.3 vs 0.7), Apo A-I/Apo B (1.67 vs. 1.85) and PON-1 (170 vs. 243 nmol/mL.min) were significantly lower in SAC than in CH, respectively. Total cholesterol (156 vs 144 mg/dL), triglycerides (TG) (119 vs. 94 mg/dL), apo A-I (133 vs. 128 mg/dL), apo B (84 vs. 73 mg/dL), hematocrit (48 vs. 41%), transferrin (295 vs. 260 mg/dL) and CETP (181 vs. 150%/mL.h) were significantly higher in SAC than in CH. There was a significant univariate association between altitude and transferrin (r0.38), hematocrit (r0.75), TG (r0.24), apo B (r0.29), PON-1 (r-0.40), and CETP (r0.37). Multiple linear regression analyses showed that altitude was significantly associated with children's TG (ß = 0.28, R2 = 0.14), HDL-C (ß = â€’0.27; R2 = 0.23), apo B (ß = 0.32; R2 = 0.14), CETP (ß = 0.38; R2 = 0.15) and PON-1 (ß = â€’0.36; R2 = 0.16), adjusted for age, gender and BMI. CONCLUSION: SAC children presented a more atherogenic lipid profile, plus lower PON1 and higher CETP activities, than CH children.

Obesity and dyslipidemia in patients with psoriasis: A case-control study.

The aim of this study was to conduct a more comprehensive analysis of the association between psoriasis and abnormal lipid metabolism.The case-control study included 222 psoriatic patients and 445 non-psoriatic control patients matched for age and gender. Clinical parameters included age, gender, and body mass index (BMI). Serum lipid levels were recorded and included cholesterol (CHO), triglycerides (TG), low-density lipoprotein (LDL), high density lipoprotein (HDL), phospholipids (PLIP), free fatty acids (FFA), lipoprotein (a) [Lp(a)], and apolipoproteins (apoA1, apoB, and apoE). Statistical analysis was carried out through the IBM Statistical Package for the Social Studies version 23.0.Compared with controls, levels of BMI and the prevalence of obesity were significantly higher in psoriatic patients. The results revealed that when compared to controls, significant elevation of serum TG (P <.001) and Lp(a) (P = .022) was observed. Levels of HDL (P <.001) and apoA1 (P <.001) were significantly lower in psoriatic patients. There was no significant difference in CHO (P = .367), LDL (P = .400), apoB (P = .294), apoE (P = .05), PLIP (P = .931) and FFA (P = .554) between patients and controls. The levels of CHO, TG, PLIP, FFA, and apoE were positively correlated with BMI level.Dyslipidemia was more common in psoriatic patients, compared with non-psoriatic controls.

Serum apolipoprotein B-to-apolipoprotein A1 ratio is independently associated with disease severity in patients with acute pancreatitis.

Early identification of severe acute pancreatitis (SAP) is critical for clinical decision-making. The apolipoprotein B-to-apolipoprotein A1 ratio (ApoB/A1 ratio) reflects the balance between pro-inflammation and anti-inflammation in vivo. This study investigated the association between serum ApoB/A1 ratio at admission and acute pancreatitis (AP) severity. A total of 375 patients with first attack of AP were retrospectively recruited from January 2014 to December 2017. The severity of AP was assessed at admission based on the 2012 revised Atlanta Classification. Serum lipids levels were tested on the first 24 h of hospitalization, of which the correlations with clinical features or scoring systems were also measured. The ApoB/A1 ratio markedly increased across disease severity of AP. The ApoB/A1 ratio, expressed as both quartile and continuous variables, was significantly associated with a high risk of SAP, even after adjustment for other conventional SAP risk factors. The ApoB/A1 ratio positively correlated with the revised 2012 Atlanta Classification, Ranson score, Bedside Index for Severity in AP score, Modified Computed Tomography Severity Index score, and Acute Physiology and Chronic Health Evaluation II score for AP severity. The optimal cut-off value of ApoB/A1 ratio for detecting SAP was 0.88, with a sensitivity of 83.08% and a specificity of 69.03%. Serum ApoB/A1 ratio at admission is closely correlated with disease severity in patients with AP and can serve as a reliable indicator for SAP in clinical setting.

Lipoprotein(a) Levels and the Risk of Myocardial Infarction Among 7 Ethnic Groups.

BACKGROUND: Lipoprotein(a) [Lp(a)] levels predict the risk of myocardial infarction (MI) in populations of European ancestry; however, few data are available for other ethnic groups. Furthermore, differences in isoform size distribution and the associated Lp(a) concentrations have not fully been characterized between ethnic groups. METHODS: We studied 6086 cases of first MI and 6857 controls from the INTERHEART study that were stratified by ethnicity and adjusted for age and sex. A total of 775 Africans, 4443 Chinese, 1352 Arabs, 1856 Europeans, 1469 Latin Americans, 1829 South Asians, and 1221 Southeast Asians were included in the study. Lp(a) concentration was measured in each participant using an assay that was insensitive to isoform size, with isoform size being assessed by Western blot in a subset of 4219 participants. RESULTS: Variations in Lp(a) concentrations and isoform size distributions were observed between populations, with Africans having the highest Lp(a) concentration (median=27.2 mg/dL) and smallest isoform size (median=24 kringle IV repeats). Chinese samples had the lowest concentration (median=7.8 mg/dL) and largest isoform sizes (median=28). Overall, high Lp(a) concentrations (>50 mg/dL) were associated with an increased risk of MI (odds ratio, 1.48; 95% CI, 1.32-1.67; P<0.001). The association was independent of established MI risk factors, including diabetes mellitus, smoking, high blood pressure, and apolipoprotein B and A ratio. An inverse association was observed between isoform size and Lp(a) concentration, which was consistent across ethnic groups. Larger isoforms tended to be associated with a lower risk of MI, but this relationship was not present after adjustment for concentration. Consistent with variations in Lp(a) concentration across populations, the population-attributable risk of high Lp(a) for MI varied from 0% in Africans to 9.5% in South Asians. CONCLUSIONS: Lp(a) concentration and isoform size varied markedly between ethnic groups. Higher Lp(a) concentrations were associated with an increased risk of MI and carried an especially high population burden in South Asians and Latin Americans. Isoform size was inversely associated with Lp(a) concentration, but did not significantly contribute to risk.

Ultrassequenciamento exômico dos principais genes relacionados com a hipercolesterolemia familial / Ultrasequensing exomic of the main genes related to familial hypercholesterolemia

A frequência de Hipercolesterolemia Familial (HF) ainda é desconhecida no Brasil, principalmente pela ausência de estudos com caracterização genotípica associada à fenotípica. Os dados epidemiológicos existentes se baseiam apenas no fenótipos e carecem do diagnóstico molecular confirmatório. O objetivo do presente estudo foi identificar as principais causas genéticas da HF em pacientes diagnosticados fenotipicamente através de um painel exômico com 61 genes a fim de contribuir para um sistema de confirmação do diagnostico molecular em uma amostra da população brasileira. Para isso foram incluídos 141 pacientes, não aparentados, portadores de HF atendidos pelo setor de dislipidemias do Instituto Dante Pazzanese de Cardiologia, Laboratório de Analises Clinicas da Faculdade de Ciências Farmacêuticas da Universidade Federal do Rio Grande do Norte e do Programa Hipercol Brasil do Instituto do Coração. As amostras de sangue periférico foram obtidas para determinações fenotípicas laboratoriais e extração de DNA genômico. A biblioteca de DNA foi construída utilizando o kit Nextera® Rapid Capture Enrichment Custom enriquecendo os éxons de 61 genes que direta ou indiretamente estão relacionados com metabolismo do colesterol. O ultrassequenciamento foi realizado utilizando kit MiSeq Reagent (300 a 500 ciclos) na plataforma MiSeq (Illumina). Os resultados de sequenciamento foram inicialmente alinhados a uma sequência referência e analisados para eliminação de falsos positivos, segundo os parâmetros de qualidade, tais como: cobertura mínima de 30x, frequência do alelo alterado maior que 20% e diferença da distribuição das leituras entre as sequências nucleotídicas menor que 15%. Foram identificadas 472 diferentes variantes em 56 dos genes presentes no painel, sendo 45 consideradas como não descritas. Nos genes APOA1, APOA2, LIPC, RBP4 e TIMP1 não foram observadas variantes dentro dos critérios estabelecidos. Das variantes observadas 25 identificadas em 30 (21,2%) pacientes já tinha sido publicadas em relação à HF nos três principais genes (LDLR, APOB e PCSK9), confirmando o diagnóstico. Foi caracterizado genotipicamente outras dislipidemias primárias em 7 pacientes, sem diagnóstico molecular de HF, através de variantes identificadas no ultrassequenciamento em outros genes. Dos 104 pacientes que não possuíam nenhuma variante já previamente caracterizada, 69 possuíam variantes relacionados com o metabolismo do colesterol. As variantes sem patogenicidade conhecida foram avaliadas através de ferramentas de predição in silico e 22 delas possuíam características sugestivas de patogenicidade em pelo menos 4 das ferramentas utilizadas, duas delas também mostraram alterar a estrutura da proteína segundo análises de docking molecular. Foram identificadas também 223 variantes em região não transcritas (UTR). Quando realizada as análises estatística de todas as variantes identificadas, observamos associação de 13 variantes com concentrações mais elevadas de colesterol da LDL, 5 com concentrações mais elevadas de apolipoproteina B-100, 5 com concentrações mais elevadas de colesterol total, 6 com presença de arco córneo, 2 com manifestação de xantelasmas, 2 com ausência de xantomas e 3 com a presença de doença arterial coronariana. Dessas 6 variantes já haviam sido previamente descritas com HF ou algum outro fenótipo associado e 2 não tinham citação na literatura pesquisada, mas possuíam característica patogênica para a proteína segundo as ferramentas de predição in silico. Este estudo permitiu a identificação das causas genéticas da HF em pacientes brasileiros diagnosticados fenotipicamente, mostrando que a técnica escolhida permitiu caracterizar 21,2% dos pacientes. Além disso, foi possível identificar outras dislipidemias primárias e caracterizar algumas variantes que, apesar de necessitarem serem validadas, indicam uma possível associação com a HF, aumentando o esclarecimento do fenótipo com o genótipo para 74,5%. Este estudo também possibilitou a identificação de novas variantes que devem ser avaliadas para confirmar associação com a doença e utilizar para o diagnóstico propondo um novo painel poligênico

The frequency of Familial Hypercholesterolemia (FH) is still unknown in Brazil, mainly due to the absence of studies with genotypic characterization associated with phenotype. Existing epidemiological data are based only on the phenotypes and lack the confirmatory molecular diagnosis. The aim of the present study was to identify main genetic causes of FH in patients diagnosed phenotypically through an exomic panel with 61 genes in order to contribute to a system of confirmation molecular diagnosis in a sample of the Brazilian population. To this end, 141 non-related patients with FH treated by the dyslipidemia sector of the Institute Dante Pazzanese of Cardiology, Clinical Analysis Laboratory of the Faculty of Pharmaceutical Sciences of the University Federal of Rio Grande do Norte and the Hipercol Brazil Program of the Heart Institute. Peripheral blood samples were obtained for laboratory phenotypic determinations and extraction of genomic DNA. The DNA library was constructed using the Nextera® Rapid Capture Enrichment Custom kit, enriching with éxons of 61 genes that are directly or indirectly related to cholesterol metabolism. Ultrasequencing was performed using MiSeq Reagent kit (300 to 500 cycles) on the MiSeq platform (Illumina). The sequencing results were initially aligned to a reference sequence and analyzed for false positive elimination according to quality parameters such as: minimum coverage of 30x, altered allele frequency greater than 20%, and difference in the distribution of reads between sequences nucleotides less than 15%. 472 different variants were identified in 56 of the genes present in the panel, of which 45 were considered not described. In the APOA1, APOA2, LIPC, RBP4 and TIMP1 genes no variants were observed within the established criteria. In 25 of the variants observed presents in 30 (21.2%) patients had already been published in relation to FH in the three main genes (LDLR, APOB and PCSK9), confirming the diagnosis. Other primary dyslipidemias were caracterized genotypically in 7 patients, without molecular diagnosis of HF, through variants identified in ultrasequencing in other genes. Of the 104 patients who did not have any previously characterized variant, 69 had variants related to cholesterol metabolism. The variants without known pathogenicity were evaluated using in silico prediction tools and 22 of them had characteristics suggestive of pathogenicity at least 4 of the tools used, two of them also showed to alter the structure of the protein according to molecular docking analyzes. Were also identified 223 non-transcribed region (UTR) variants. Statistical analysis of all the variants identified showed association of 13 variants with higher concentrations of LDL cholesterol, 5 with higher concentrations of apolipoprotein B-100, 5 with higher concentrations of total cholesterol, 6 with presence of an arc corneal, 2 with manifestation of xanthelasms, 2 with absence of xanthomas and 3 with the presence of coronary artery disease. Of these 6 variants had previously been described with HF or some other associated phenotype and 2 had no citation in the researched literature, but had a pathogenic characteristic for the protein according to in silico prediction tools. This study allowed the identification of the genetic causes of FH in Brazilian patients diagnosed phenotypically, showing that the technique chosen allowed to characterize 21.2% of the patients. In addition, it was possible to identify other primary dyslipidemias and to characterize some variants that, although they need to be validated, indicate a possible association with HF, increasing the clarification of the phenotype with the genotype to 74.5%. This study also allowed the identification of new variants that should be evaluated to confirm association with the disease and to use for the diagnosis proposing a new polygenic panel

N-Glycoproteomic Analysis of Chicken Egg Yolk.

Posttranslational N-glycosylation of food proteins plays a critical role in their structure and function. However, the N-glycoproteome of chicken egg yolk (CEY) has not been studied yet. Glycopeptides hydrolyzed from CEY proteins were enriched, with deglycosylation occurring using PNGase F, and then were identified using a shotgun glycoproteomics strategy. A total of 217 N-glycosylation sites and 86 glycoproteins were identified in CEY, and these glycoproteins are mainly involved in the binding, biological regulation, catalytic activity, and metabolic processes. Among the identified CEY glycoproteins, 22 were recognized as proteases and protease inhibitors, suggesting that a proteinase/inhibitor regulation system exists in CEY; further, 15 were members of the complement and immune systems, which provide protection against potential threats during hatching. The study provides important structural information about CEY glycoproteins and aids in the understanding of the underlying mechanism of embryo development as well as changes in CEY functional characteristics during storage and processing.

Update on the laboratory investigation of dyslipidemias.

The role of the clinical laboratory is evolving to provide more information to clinicians to assess cardiovascular disease (CVD) risk and target therapy more effectively. Current routine methods to measure LDL-cholesterol (LDL-C), the Friedewald calculation, ultracentrifugation, electrophoresis and homogeneous direct methods have established limitations. Studies suggest that LDL and HDL size or particle concentration are alternative methods to predict future CVD risk. At this time there is no consensus role for lipoprotein particle or subclasses in CVD risk assessment. LDL and HDL particle concentration are measured by several methods, namely gradient gel electrophoresis, ultracentrifugation-vertical auto profile, nuclear magnetic resonance and ion mobility. It has been suggested that HDL functional assays may be better predictors of CVD risk. To assess the issue of lipoprotein subclasses/particles and HDL function as potential CVD risk markers robust, simple, validated analytical methods are required. In patients with small dense LDL particles, even a perfect measure of LDL-C will not reflect LDL particle concentration. Non-HDL-C is an alternative measurement and includes VLDL and CM remnant cholesterol and LDL-C. However, apolipoprotein B measurement may more accurately reflect LDL particle numbers. Non-fasting lipid measurements have many practical advantages. Defining thresholds for treatment with new measurements of CVD risk remain a challenge. In families with genetic variants, ApoCIII and lipoprotein (a) may be additional risk factors. Recognition of familial causes of dyslipidemias and diagnosis in childhood will result in early treatment. This review discusses the limitations in current laboratory technologies to predict CVD risk and reviews the evidence for emergent approaches using newer biomarkers in clinical practice.

Comparison of fasting and non-fasting lipid profiles in a large cohort of patients presenting at a community hospital.

OBJECTIVE: To compare the fasting and non-fasting lipid profile including ApoB in a cohort of patients from a community setting. Our purpose was to determine the proportion of results that could be explained by the known biological variation in the fasting state and to examine the additional impact of non-fasting on these same lipid parameters. METHODS: 1093 adult outpatients with fasting lipid requests were recruited from February to September 2016 at the blood collection sites of the Moncton Hospital. Participants were asked to come back in the next 3-4days after having eaten a regular breakfast to have their blood drawn for a non-fasting lipid profile. RESULTS: 91.6% of patients in this study had a change in total cholesterol that fell within the biological variation expected for this parameter. Similar results were seen for HDL-C (94.3%) non-HDL-C (88.8%) and ApoB (93.0%). A smaller number of patients fell within the biological variation expected for TG (78.8%) and LDL-C (74.6%). An average TG increase of 0.3mmol/L was observed in fed patients no matter the level of fasting TG. A gradual widening in the range of change in TG concentration was observed as fasting TG increased. Similar results were seen in diabetic patients. CONCLUSION: Outside of LDL-C and TG, little changes were seen in lipid parameters in the postprandial state. A large part of these changes could be explained by the biological variation. We observed a gradual widening in the range of increase in TG for patients with higher fasting TG. Non-HDL-C and ApoB should be the treatment target of choice for patients in the non-fasting state.

Changes in apolipoprotein B and oxidized low-density lipoprotein levels in gingival crevicular fluids as a result of periodontal tissue conditions.

BACKGROUND AND OBJECTIVE: Periodontal disease is a chronic inflammatory disease caused by bacterial infection that can lead to tooth loss. Gingival crevicular fluid can be collected easily and noninvasively. We previously discovered the presence of apolipoprotein B (apoB), the main constituent of low-density lipoprotein, and oxidized low-density lipoprotein (oxLDL) in the gingival crevicular fluid of healthy subjects. In this study, we investigated whether periodontal conditions affect the levels of apoB and oxLDL in gingival crevicular fluid. MATERIAL AND METHODS: The study population comprised 11 patients with chronic periodontitis. A pair of gingival crevicular fluid samples was collected from each patient at a healthy site and at a site with periodontitis (baseline samples). Thereafter, gingival crevicular fluid samples were collected from the same patients again at 4 and 8 wk after scaling and root planing (SRP). The levels of apoB, oxLDL, protein and cytokines in gingival crevicular fluid, in addition to gingival crevicular fluid volume, were measured. RESULTS: At baseline, the levels of apoB and oxLDL in gingival crevicular fluid were higher at the sites with periodontitis than at the healthy sites. The levels of apoB and oxLDL at periodontal sites decreased after SRP. The level of oxLDL in gingival crevicular fluid correlated well with the probing pocket depth. The oxLDL : apoB ratio in gingival crevicular fluid was significantly higher than that in plasma. CONCLUSIONS: The levels of apoB and oxLDL in gingival crevicular fluid change according to the periodontal tissue conditions.