Targeted sequencing of a gene panel in patients with familial hypercholesterolemia from Southern Poland.

INTRODUCTION: Familial hypercholesterolemia (FH) is an autosomal dominant monogenic lipid metabolism disorder characterized by a significantly elevated level of low­density lipoprotein (LDL) cholesterol and leading to premature ischemic heart disease. FH is caused by mutations in the LDLR, APOB, and PCSK9 genes; however, these mutations account for only about 40% of FH cases. In order to obtain a genetic diagnosis of FH, sequencing of other genes involved in the lipid metabolism might be useful. OBJECTIVES: This study aimed to describe genetic variants in genes associated with FH in a group of patients from the Malopolska province in Southern Poland, using the targeted next generation sequencing (NGS) technology. PATIENTS AND METHODS: The study involved 90 unrelated adults (age range, 18-70 years) with FH diagnosed clinically according to the Simon Broome Register criteria. A custom­designed capture assay and the Illumina MiSeq platform were used. The panel included exons and exon / intron boundaries of known FH­causing genes: LDLR, APOB, and PCSK9, as well as genes previously associated with high cholesterol levels: APOE, ABCG5, ABCG8, LPL, NPC1, LDLRAP1, LIPC, STAP1, and CELSR2. Genetic variants were classified based on in silico predictions and ClinVar reports. RESULTS: We detected 4 patients with variants in the LDLR and APOB genes that had not been previously linked to FH in ClinVar. We also found APOB mutations outside the common LDL receptor-binding region, in exons 26 and 29. Interestingly, we observed a high frequency of pathogenic variants in exon 4 of the APOE gene: rs7412, probably damaging (4 patients) and rs429358, benign (16 patients). CONCLUSIONS: NGS is a useful and reliable method to detect new variants in genes related to FH. In addition, the results enable the detection of FH phenocopies and introduction of appropriate treatment.

Serum PCSK9 Correlates with PTX3 and Apolipoproteins B, A1, and C3 Concentrations in Patients with Type 2 Diabetes.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is involved in the regulation of LDL metabolism. There is evidence that circulating PCSK9 is a cardiovascular risk factor. In this study, we determined factors associated with circulating PCSK9 in a group of patients with type 2 diabetes mellitus (DM2). Material included 116 consecutive patients with DM2 from outpatient diabetes clinic. Circulating PCSK9, PTX3, apolipoprotein (apo) B100, apo B48, and apo C3 levels were determined by ELISA, apo A1 by immunoturbidimetry. The mean (sd) age of patients was 59.1 (11.1) years, the mean (sd) values of serum PCSK9 were 255.4 (106.97) ng/ml. Circulating PCSK9 correlated negatively with age (r = -0.21, p < 0.05) and HbA1c (r = -0.21, p < 0.05) and positively with BMI (r = 0.21, p < 0.05), total cholesterol (r = 0.59), LDL-cholesterol (r = 0.50), triglyceride (r = 0.35), apo B100 (r = 0.43), apo A1 (r = 0.43) (p < 0.001 for all), apo C3 (r = 0.29, p < 0.01), and apo B48 (r = 0.25, p < 0.01) concentration and FLI (r = 0.26, p < 0.01). Strong correlation between PTX3 and PCSK9 levels was observed (r = 0.47, p < 0.001). Multiple stepwise backward regression analysis with PCSK9 as dependent variable revealed that PTX3, apo B100, apo A1, apo B48, and BMI were significantly positive and the presence of NAFLD and HbA1c negatively associated with PCSK9 concentrations. These variables together explain 57% of PCSK9 variability; the strongest relationship was observed between PCSK9 and PTX3 and apo B100. Our results indicate that circulating PCSK9 is significantly associated with inflammation marker PTX3 as well as atherogenic lipids and apolipoproteins C3, B100, and B48, which might be of value in understanding interactions between development of atherosclerosis and inflammatory state in DM2 patients.

Correlates of pentraxin 3 serum concentration in men and women with type 2 diabetes.

Elevated levels of plasma pentraxin 3 (PTX3), a marker of inflammation, are associated with the risk of developing cardiovascular diseases in the general population, as well as in patients with type 2 diabetes (DM2). In this study, we aimed to determine factors associated with PTX3 serum concentrations in men and women with DM2. The study included 116 consecutive patients (67 men and 49 women) with DM2 from an outpatient diabetic clinic. Men were characterised by lower age and higher uric acid, creatinine and bilirubin concentrations and waist/hip ratio than women. In women, low-density lipoprotein cholesterol (LDL-C) levels were higher than in men. In men, median (interquartile range) values of PTX3 concentration were 4.02 (1.99), and in women they were 4.53 (3.31) ng/ml (NS). In men, PTX3 concentrations correlated with total cholesterol (TC), triglycerides, apolipoprotein (Apo) C3, Apo B48, Glc and creatinine levels. In women, PTX3 correlated significantly with TC and LDL-C and Apo B100. Partial regression analysis revealed that after adjusting for age, PTX3 concentrations in men were significantly associated with TC, LDL-C, triglycerides, creatinine, Apo C3 and Apo B48, while in women they were associated with TC, LDL-C and Apo B100. The results could be of importance in sex-specific prevention of vascular complications in DM2 patients.

Serum pentraxin 3 concentration in patients with type 2 diabetes and nonalcoholic fatty liver disease.

INTRODUCTION: Nonalcoholic fatty liver disease (NAFLD) is common in patients with type 2 diabetes (T2D). Pentraxin 3 (PTX3), a marker of inflammation, is a cardiovascular risk factor. OBJECTIVES: We examined clinical and biochemical factors associated with serum PTX3 concentrations in patients with T2D with and without NAFLD. PATIENTS AND METHODS: Serum material was obtained from 116 patients with T2D (mean age, 59.1 years), including 79 patients with NAFLD. RESULTS: Median (interquartile range) PTX3 level was 4.264 (2.293) ng/ml in patients with and 3.773 (3.223) ng/ml in patients without NAFLD (P = 0.93). In the whole group, PTX3 level was associated with total cholesterol, low­density lipoprotein cholesterol (LDL­C), apolipoprotein (apo) B100, apo C3, triglyceride (TG) concentrations, and waist circumference after adjustment for age and gender. As indicated by partial regression coefficient b, increase of independent variable LDL­C by 1 mmol/l was associated with the rise of PTX3 by 1.2017 ng/ml, increase of apo B100 by 1 mg/dl with the rise of PTX3 by 1.0051 ng/ml, and increase of apo C3 by 1 µg/dl with the rise of PTX3 by 1.0012 ng/ml. In patients with T2D with NAFLD, total cholesterol, LDL­C, TG, apo C3, and apo B100 were associated with PTX3. Associations of PTX3 with apolipoproteins were observed only in the NAFLD group. CONCLUSIONS: Reported associations of PTX3 level add new insight into possible mechanisms of its atherogenic actions.

[Psychosocial determinants of smoking]. / Psychospoleczne determinanty nalogu palenia papierosów.

Prevalence of smoking is high in the middle age Polish population despite of wide spread knowledge on health hazards. The goal of the paper was to assess the relation between smoking with awareness of smoking hazards and psychosocial factors related to increased risk of coronary heart disease. The studied group was a random sample of residents of Cracow aged between 45-64 years. Out of 1756 men and 1788 women, selected were 1154 men and 1156 women (65%) which participated in the study. In the studied group 40% of men and 20% of women were smokers. Over 95% of people knew that smoking is a risk factor of myocardial infarction. However 15% shared the opinion that quitting smoking is also a risk factor of myocardial infarction. The latter opinion was more frequent in persons with low education and in smokers. Smoking men and women were more depressive. There were some differences in relations between smoking and other psychosocial factors studied between men and women. In men, smokers reported higher effort and lower reward at work. Perceived general control and control over life were related to smoking in women. In persons for whom health argument is not sufficient to refrain from smoking or quit smoking, the effectiveness of anti-smoking program might depend on relating it to depressive symptoms and on applying different motivation in men and women by depending on psychosocial characteristics.

[Cardiovascular autonomic neuropathy and nephropathy in patients with type-1 diabetes mellitus]. / Neuropatia autonomiczna ukladu sercowo- naczyniowego oraz powiklania nerkowe w cukrzycy typu 1.

UNLABELLED: In the last few years much attention is being given to the problem of diabetes mellitus, to it's development, prevalence and progression of chronic complications. The aim of the study was to 1. evaluate correction of metabolic disturbances in patients with long-term type 1 diabetes mellitus; 2. evaluate occurrence of diabetic nephropathy and excretion function in patients with long-term type 1 diabetes mellitus; 3. evaluate function of the cardiovascular autonomic nervous system in patients with long-term type 1 diabetes mellitus; 4. search for connections between the type and change dynamics in the cardiovascular system and degree of diabetic nephropathy advancement. The study was performed in a group consisting of 31 patients with type 1 diabetes mellitus (20 women, 11 males). Mean age of the study group equaled 37.6 +/- 10.75 years, duration of diabetes mellitus 21.3 +/- 9.55 years. Concentration of cystatin C in the study group was 0.98 +/- 0.23 ng/ml, in the group of patients with diabetic retinopathy 1.13 +/- 0.30 ng/ml, while in the group without diabetic retinopathy 0.89 +/- 0.13 ng/ml. Concentration of cystatin C (p <0.05) and calculated GFR according to equations proposed by F.J. Hoeck [GFR/1.73 m2 = -4.32 + 80.3/plasma cystatin] (p < 0.01) also G.D. Tan [GFR -10 = (87.1/plasma cystine) - 6.87] (p < 0.01) significantly differentiated the discussed groups. CONCLUSIONS: despite normal levels of blood creatinine in the studied patients, decreased glomerular filtration was calculated for plasma cystatin C. Plasma cystatin C concentrations and calculated glomerular clearance significantly differentiated the group with retinopathy from the group without diabetic retinopathy. Determining cystatin C concentrations as a protein which probably does not undergo glycation in plasma, may play a role in the detection of early diabetic nephropathy, when as well as plasma creatinine levels and albumin/creatinine index calculated from a sample of morning urine do not differentiate the studied group of patients.

Lifestyle characteristics and hypertension in the middle-aged population of Kraków.

Hypertension affects nearly one-third of total Polish adult population. Poor control of hypertension largely contributes to high cardiovascular disease (CVD) mortality in Poland. However, satisfactory hypertension control at the national level could be beyond economical conditions. Prevention of hypertension by lifestyle modification may be one of the ways to decrease CVD population risk attributed to hypertension. The goal was to assess the relation between the chance of having hypertension and lifestyle characteristics, i.e. education, smoking, physical activity, salt consumption, ethanol consumption and body mass index (BMI) in the middle-aged population of the urban population of Kraków. A cross-sectional study of a random sample of 1157 women and 1153 men, permanent inhabitants of Kraków at age 45-64 years, was carried out by home interview followed by a visit in the clinic. There were 66% men and 56% women who had hypertension. After adjustment for age, BMI over 27.5 kg/m2 in men and over 25.0 kg/m2 in women was strongly related to hypertension. In men, odds ratio (OR) increased from 1.96 in the group with overweight (27.5 kg/m2 < or = BMI < 30.0 kg/m2) to 5.33 in the group with obesity (BMI > or = 35.0 kg/m2). In women, OR increased from 1.6 in the group with overweight (25.0 kg/m2 < or = BMI < 27.5 kg/m2) to 13.42 in the group with obesity (BMI > or = 35.0 kg/m2). In men, after adjustment for age and BMI, a higher chance of hypertension was found for binge drinkers but the result was of borderline significance (p = 0.057). In women, moderate alcohol consumption was related to lower chance of hypertension (OR = 0.72). Women who were current smokers had lower chance of hypertension (OR = 0.67). No significant relation between smoking and hypertension was found in men. After adjustment for age and BMI, there was no significant relation between hypertension and physical activity. However, both in men and in women, there was a dose effect of leisure time physical activity on the average estimate of the chance of hypertension. Education, estimate of sodium consumption and blood total cholesterol (TC) were not related to hypertension. The findings confirmed that weight control may be a strong tool for preventing hypertension and increasing physical activity and refraining from binge drinking may have a protective effect.