Molecular characterization of Polish patients with familial hypercholesterolemia: novel and recurrent LDLR mutations.

Autosomal dominant hypercholesterolemia (ADH) is caused by mutations in the genes coding for the low-density lipoprotein receptor (LDLR), apolipoprotein B-100 (APOB), or proprotein convertase subtilisin/kexin type 9 (PCSK9). In this study, a molecular analysis of LDLR and APOB was performed in a group of 378 unrelated ADH patients, to explore the mutation spectrum that causes hypercholesterolemia in Poland. All patients were clinically diagnosed with ADH according to a uniform protocol and internationally accepted WHO criteria. Mutational analysis included all exons, exon-intron boundaries and the promoter sequence of the LDLR, and a fragment of exon 26 of APOB. Additionally, the MLPA technique was applied to detect rearrangements within LDLR. In total, 100 sequence variations were identified in 234 (62%) patients. Within LDLR, 40 novel and 59 previously described sequence variations were detected. Of the 99 LDLR sequence variations, 71 may be pathogenic mutations. The most frequent LDLR alteration was a point mutation p.G592E detected in 38 (10%) patients, followed by duplication of exons 4-8 found in 16 individuals (4.2%). Twenty-five cases (6.6%) demonstrated the p.R3527Q mutation of APOB. Our findings imply that major rearrangements of the LDLR gene as well as 2 point mutations (p.G592E in LDLR and p.R3527Q in APOB) are frequent causes of ADH in Poland. However, the heterogeneity of LDLR mutations detected in the studied group confirms the requirement for complex molecular studies of Polish ADH patients.

Clinical significance of apoptotic index in non-small cell lung cancer: correlation with p53, mdm2, pRb and p21WAF1/CIP1 protein expression.

PURPOSE: The aim of this study was to assess the prognostic relevance of apoptotic index (AI), considered alone or together with expression of several proteins controlling G1 check point (p53, mdm2, pRb and p21WAF1/CIP1) in non-small cell lung cancer (NSCLC) patients. METHODS: Study group included 50 NSCLC patients who underwent curative pulmonary resection. Apoptosis was detected with the use of TUNEL technique and AI was defined as the number of apoptotic cells per 1,000 tumor cells. The expression of p53, mdm2, pRb and p21WAF1/CIP1 was assessed immunohistochemically. RESULTS: The mean and median AI calculated for all 50 patients was 14 and 9, respectively. Patients with lower (<14) and higher (> or =14) AI constituted 35 (70%) and 15 (30%) of cases, respectively. AI was not correlated with patient clinical characteristics, and expression of p53, pRb and p21WAF1/CIP1 . However, lower AI was correlated with over-expression of mdm2 protein (P=0.04). Median survival for patients with lower and higher AI was 43 months and 22 months, respectively, and 5-year survival probability-60 and 25%, respectively (P=0.03). In multivariate analysis, the only variable associated with shortened survival was AI (P=0.03, HR=2.9, 95% CI 1.95-3.86). CONCLUSIONS: These results suggest that AI correlates with mdm2 protein expression and influences survival in NSCLC.

Relationship between fasting plasma insulin and ambulatory blood pressure in young, non-obese normotensive subjects.

In order to investigate the relationship between ambulatory blood pressure values and fasting plasma insulin, non-invasive, 24-h blood pressure monitoring was performed in 32 young normotensive males. Systolic and diastolic blood pressures were averaged for awake and asleep periods. Fasting plasma insulin levels correlated significantly with both asleep (r = 0.61; P less than 0.001) and awake (r = 0.44; P less than 0.02) systolic blood pressure, but not with casual systolic blood pressure (r = 0.27). There were no significant associations between awake, asleep and casual diastolic blood pressure, and fasting plasma insulin levels (r = 0.15, 0.05 and 0.21, respectively). These results support the hypothesis that insulin may be a physiological determinant of blood pressure.

Genetic influences on insulinemia in normotensive twins.

The aim of this study was to establish the contribution of genetic factors to the variance of plasma insulin concentration in healthy, normotensive twins. Seventeen pairs of monozygotic (MZ) and 17 pairs of dizygotic (DZ) twins were investigated. The test of genetic variance revealed a significantly larger within-pair variance of fasting plasma insulin (FPI) and a relative insulin resistance (RIR) in the DZ twins, in comparison with the MZ twins. Both FPI and RIR had a higher intraclass correlation coefficient in the MZ twins than in the DZ twins; the corresponding heritability estimates were 0.54 for FPI and 0.66 for RIR. Adjusting for age, gender, and body mass index did not affect heritability estimates for either FPI or RIR. Our data indicate that genetic factors are important determinants of insulinemia in normal subjects, independent of body mass index.

Awareness and treatment of arterial hypertension in Poland.

In order to find out the general public's opinion on blood pressure (BP) control in Poland a poll was conducted on a representative sample of 2080 randomly selected subjects. Overall 71% of the subjects were aware of their BP level. The awareness of BP level increased progressively with age and education level. Among those who were aware of their BP level, 21% had hypertension. Only 43% of hypertensives were treated on a regular basis. Cigarettes were smoked by 33% of the patients. Only 16% of the hypertensives used a low-salt diet, and only 31% tried to reduce animal fat in their diet. More than a half of the hypertensives did not adjust their lifestyle and daily habits to the disease. In conclusion, these results indicate that the prevention and treatment of hypertension in Poland should be urgently improved.

[Spectrum analysis of heart rate. A method of studying the role of the autonomic nervous system in the regulation of blood circulation]. / Analiza widma czestosci akcji serca. Metoda badania udzialu ukladu autonomicznego w regulacji krazenia.

Spectral analysis of heart rate variability has recently been shown to be a reliable noninvasive test for quantitative assessment of cardiovascular automatic regulatory responses. In 12 ambulant normotensive healthy young males (mean age 23 +/- 1 years) after a period of 10 min. for stabilisation, a continuous ecg recording (lead CM-5) for 8 min. was obtained in the supine and standing position, with a controlled respiration rate 15/min. Power spectrum of 512 point time series (R-R intervals) in both positions was calculated using a fast Fourier transform-based window periodogram method. Based upon results from the literature the power spectrum analysis was performed on two components: low frequency LF (0.05-0.15 Hz) and high frequency HF (0.15-0.50 Hz). Mean R-R interval decreased on standing position from 0.79 +/- 0.10 s to 0.59 +/- 0.11 s (p less than 0.001). The ratio HF/LF in supine was 0.63 +/- 0.70 and on standing position 2.54 +/- 0.73 (p less than 0.001). The relative LF component of the total HR power spectrum increased from 22.8% +/- 12.1% to 42.9 +/- 14.4% (p less than 0.001) after changing the position from supine to standing, and the relative HF component decreased from 56.3 +/- 22.4% to 25.5 +/- 16.2 (p less than 0.001). The total power was significantly lower when standing in comparison to supine position (681 +/- 519 s2, 1188 +/- 963 s2 respectively, p less than 0.05). Our results suggest that heart rate fluctuations in supine position in normal men are mainly vagally determined (HF power spectrum component).(ABSTRACT TRUNCATED AT 250 WORDS)

[Effects of isometric exercise on left ventricular function assessed with polycardiography in patients with early type I diabetes]. / Wplyw wysilku izometrycznego na czynnosc lewej komory serca oceniana metoda polikardiograficzna u chorych z krótkotrwajaca cukrzyca typu I.

15 patients with controlled type 1 diabetes lasting up to 24 months underwent the study. The control group consisted of 22 healthy men. Noninvasive examinations were carried out at rest and during a 3-minute isometric exercise of a load equal to 30% maximal effort. Rest systolic and diastolic blood pressures (SBP and DBP) were almost the same in both groups. Also exercise peak SBP and DBP did not significantly differ in examined groups. Rest PEP/LVET ratio were significantly lower in diabetics (means = 0.313 vs means = 0.348 in the control group; p less than 0.002). Peak isometric exercise PEP/LVET ratio significantly increased to 0.333 in diabetics, comparing with its rest value, whereas significantly decreased to 0.333 in the control group. Results indicate increased resting left ventricular contractility in patients with early, type 1 diabetes and impairment of a left ventricular adaptation for the isometric exercise in comparison with the control group.

[Echocardiographic assessment of left ventricular function in patients with early type I diabetes]. / Echokardiograficzna ocena funkcji lewej komory serca u pacjentów z wczesna cukrzyca typu I.

M-mode, resting echocardiographic examination has been performed under 2-D control in 19 patients (mean age-26 years) with controlled, type 1 diabetes lasting up to 2 years (D group). The control group (C) consisted of 20 healthy men (average age-25 yrs). Greater cardiac index has been stated in D group (p less than 0.05). Left ventricular systolic function parameters such as: ejection fraction, percentage of fractional shortening and a mean rate of circumferential shortening indicate greater contractility in D patients. Left ventricular mass also has been significantly greater (p less than 0.04) and the total peripheral resistance significantly lowered (p less than 0.05) in the D group. Stated alterations in left ventricular function and structure are probably due to adaptation for increased peripheral flow.

[Autonomic regulation of the circulatory system in early type I diabetes]. / Odruchowa regulacja ukladu krazenia w krótkotrwajacej cukrzycy typu I.

In the group of 16 men with controlled, type I diabetes lasting up to 24 months, simple tests for assessment of autonomic regulation of the circulatory system such as a deep breathing test, Valsalva's manoeuvre and a tilt trial have been performed. Variations of systolic and diastolic blood pressures as well as heart rate have been studied. The control group consisted of 22 young, healthy men. Rest heart rate has been significantly increased prior the all tests but its alterations during deep breathing have been significantly decreased in diabetics (EM index mean = 1.44 vs 1.74 in the control group; p less than 0.01). Also Valsalva's index has been significantly increased in them (1.67 vs 1.42; p less than 0.02). Significant reduction of a delay index at the early tilt test period has been stated in diabetics (1.31 vs 1.57; p less than 0.01) as well as significant lowering of diastolic blood pressure in 1st and 5th minute of a tilt test. Performed tests indicate that even short duration of diabetes changes the autonomic regulation of the circulatory system due to inequality of para- and sympathetic system tones and/or the early autonomic neuropathy.

[Circadian rhythm of blood pressure and heart rate in hypertension with type 2 diabetes mellitus]. / Rytm dobowy cisnienia tetniczego i tetna w nadcisnieniu tetniczym i curkzycy typu 2.

Hypertension is an important risk factor for cardiovascular complications of diabetes. Most of the studies performed in diabetics so far, however, have dealt with the assessment of blood pressure by traditional sphygmomanometry. In order to investigate the circadian pattern of blood pressure and heart rate in patients with different categories of glucose tolerance, we performed ambulatory blood pressure monitoring in 28 obese hypertensives without clinical nephropathy divided in two groups. Group A consisted of 14 hypertensive males with type 2 diabetes mellitus (mean age 49.7 +/- 7.1 years, mean duration of diabetes 4.0 +/- 2.9 years); group B consisted of 14 hypertensive males with normal glucose tolerance according to National Diabetes Data Group (mean age 47.2 +/- 7.1 years). There was no significant difference in casual blood pressure (151.4/104.8 in group A versus 148.5/104.2 mmHg in group B). Ambulatory blood pressure monitoring revealed significantly higher systolic blood pressure in group A during the day (162.2 +/- 12.1 vs 152.1 +/- 9.0 mmHg in group B, P < 0.05) and at night (141.0 +/- 13.2 vs 125.5 +/- 12.5 mmHg in group B, P < 0.005). That suggests that casual readings underestimate systolic blood pressure as a predictor for macrovascular events in type 2 diabetics. The decline in nocturnal heart rate was significantly lower in group A (11.2 +/- 5.2 min-1) in comparison to group B (16.9 +/- 7.0 min-1; P < 0.05) suggesting reduced parasympathetic tone at night in diabetic patients. We conclude that type 2 diabetes has significant influence on systolic blood pressure and heart rate 24-h profiles even in patients without diabetic nephropathy.

Ambulatory blood pressure and insulinaemia in different clinical conditions.

The aim of our study was to examine the relation between insulin and ambulatory blood pressure (ABPM) in several clinical conditions. ABPM monitoring was performed with Spacelabs device. Fasting plasma insulin (FPI) was measured by radioimmunoassay. In young, non-obese normotensive subjects (n = 32) there were correlations between FPI and both asleep (r = 0.61, p < 0.001), and awake systolic ABPM (r = 0.44, p < 0.01). We have not observed any significant correlation between FPI and ABP in borderline hypertensives or in patients with established hypertension. In a group of 21 hypertensive type 2 diabetics (age 52 +/- 8 years) there was no significant correlation between FPI and ABPM. Among 14 normotensive type 1 diabetics (age 31 +/- 7 years, diabetes duration > 10 years) there was a significant negative correlation between the daily dose of insulin and 24-h systolic ABPM (r =-0.63, p < 0.02). In 20 patients with renal failure on chronic haemodialysis we have found a significant negative correlation between FPI and 24-h systolic APBM (r = 0.80, p < 0.001) and 24-h diastolic ABPM (r = -0.55, p < 0.05). Similar negative correlations were found in 20 nondialysed subjects with moderate chronic renal failure. Taken together, our results suggest that insulin could reveal its hypertensive or vasodilatory effect which depends on a clinical condition of the studied subjects.

Systolic and diastolic time intervals in young diabetics.

Systolic and diastolic time intervals were used to examine left ventricular performance in 22 young diabetic men (mean age 25 years) with no apparent clinical heart disease. Pre-ejection period index (PEPI), left ventricular ejection time index (LVETI), electromechanical systole index (QS2I), PEP to LVET ratio, the a wave percentage amplitude of the apexcardiogram (a/H5 ratio), the rapid filling wave (RFW), and the A2O interval were obtained in the conventional manner in 22 diabetics and 22 healthy men. The heart rate, diastolic pressure, PEP/LVET ratio, a/H per cent ratio, and A2O interval were significantly increased and LVET decreased in the diabetic group. QS2I, PEPI, and RFW did not differ from that in the normal group. Twenty-three per cent of patients had an abnormal systolic time interval, 54 percent an abnormal diastolic time interval, and 23 per cent had both abnormal intervals. Though these studies provide no difinite evidence of a cause, the abnormalities found may reflect a subclinical diabetic cardiopathy.

[Levels of insulin in serum during fasting and circadian blood pressure profile in young health men]. / Stezenie insuliny w osoczu na czczo a profil dobowy cisnienia tetniczego u mlodych zdrowych mezczyzn.

In order to investigate the relation of ambulatory blood pressure values to fasting plasma insulin, non-invasive 24-hour blood pressure monitoring was performed in 32 young normotensive males. Systolic and diastolic blood pressures were averaged for awake and asleep periods. Fasting plasma insulin levels correlated significantly with both asleep (r = 0.61; p < 0.001) and awake systolic blood pressures (r = 0.44; p < 0.02) but not with casual systolic blood pressure (r = 0.27). There were no significant associations of awake, asleep and causal diastolic blood pressures values with fasting plasma insulin levels (r = 0.15, 0.05 and 0.21, respectively). These results support the hypothesis that insulin may be a physiological determinant of blood pressure.