Cardiovascular Research Oral Presentation Senior Scientists’ Forum
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2019-06-18T14:58:22Z 2019-06-18T14:58:22Z 2011 Cardiovascular Research Oral Presentation Senior Scientists’ Forum // Вiopolymers and Cell. — 2011. — Т. 27, № 2, доп. — С. 61-81. — англ. 0233-7657 https://nasplib.isofts.kiev.ua/handle/123456789/156502 en Інститут молекулярної біології і генетики НАН України Вiopolymers and Cell Abstracts for RECOOP Research Networks' Scientific Review & Multidisciplinary Conference April 8-9 Cardiovascular Research Oral Presentation Senior Scientists’ Forum Article published earlier |
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61
Cardiovascular Research Oral
Presentation
Senior Scientists’ Forum
62
The role of sex specific stress response in heart failure
1Heffer Marija, 2Zibar Lada
1 Department of Medical Biology, School of Medicine, Josip Juraj Strossmayer University of Osijek,
4, Huttlerova 31 000 Osijek, Croatia
2 Department of Pathophysiology, School of Medicine, Josip Juraj Strossmayer University of Osijek,
4, Huttlerova 31 000 Osijek, Croatia
mheffer@mefos.hr
Background: The stress response is an expensive multi-system effort of the organism to
maintain homeostasis. The sympathetic surge of norepinephrine and the adrenergic surge
of epinephrine converge on the adrenergic receptors of target organs. Any overwhelming
excitation may lead to various changes in the receptor expression: down-regulation,
desensitization, internalization, phosphorylation and relocation. The initial changes are
adaptive and reversible, but chronic stress leads to maladaptive remodeling and a
permanent shift in metabolic regulation.
Objectives: Human and animal studies of stress response demonstrate a sex specific
difference in central and peripheral reaction. We summarized the findings from the
existing studies connecting stress and heart failure.
Results: In both sexes stress is linked to the development of obesity, diabetes and heart
failure, but the progression of symptoms is delayed in women and connected with
reproductive hormones. A direct link between stress and heart dysfunction is established
in the case of Takotsubo cardiomyopathy. To date the development of cardiovascular
disease has been associated with the following psychosocial risk factors: depression
(more frequent in women), anxiety (more frequent in men), character traits (D
personality), chronic life stress (from work stress up to PTSD) and social isolation (low
social support). The major adverse outcomes of chronically activated hypothalamic-
pituitary-adrenal axis are arterial hypertension, atherogenic effect, metabolic changes and
increase in blood coagulation diathesis. Individual differences in the development of
cardio vascular disease include: genetic background (race, ethnicity), lifestyle habits, sex
and social status. The psychosocial stressors studied in primates require a long time of
verification, complex design and relatively small groups of animals. It is hard to replicate
the typical psychosocial context on the rodent model.
Conclusion: The animal studies of pathological changes triggered by stress continue to
suffer from a lack of a reliable stress marker, divergent findings in time-frame required
for the development of measurable symptoms and a strict distinction between sex specific
ageing and sex specific pathology.
63
The role of connexin 37 gene polymorphism (1019C>T; Pro319Ser)
in cardiovascular disease.
Pitha Jan, 1Pithova Pavlina, Hubacek Jaroslav Alois
Institute for Clinical and Experimental Medicine,
Videnska 1952/8, 140 21, Praha 4, Prague, Czech Republic
1Teaching Hospital Motol, 2nd Medical School, Department of Medicine,
V uvalu 84, 150 06, Praha 5, Prague, Czech Republic
japi@medicon.cz
In spite of the strong prognostic value of all traditional cardiovascular risk factors, still
striking differences exist in the prevalence of clinical events between patients at
apparently similar risk. One of the main reasons is different genetic background. One of
recently discussed candidate genes for cardiovascular disease is the gene for the protein
Connexin 37 (Cx37). This protein is a part of gap junctions responsible for
communications between cells including cells in the vessel wall. Studies focused on the
association between Cx37 gene polymorphism (1019C>T; Pro319Ser) and cardiovascular
disease demonstrates inconsistent results. Our findings in 1,316 men and women
indicated that the Cx37 gene polymorphism (genotype CC) is significantly associated
with acute coronary syndrome in non-smoking women. In addition, in urban and rural
women from general population (n=1,056) with impaired fasting glycaemia the same
genotype is associated with increased intima media thickness of carotid arteries measured
by ultrasound. Finally, in 289 women with diabetes type 1 or 2, and in 208 women from
general population with central obesity, the CC genotype was associated with lower ankle
brachial blood pressure index. These data indicate that Cx37 gene polymorphism could
have gender- and smoking- dependent effects on acute coronary events and glucose
dependent effect on atherosclerosis in women.
64
Cardiovascular Research Oral
Presentation
Young Scientists’ Forum
65
Comparison of cardiovascular risk factors in women and men
patients with ischaemic heart disease – a retrospective study
Durkó András, Papp Zoltán, Édes István, Borbély Attila
University of Debrecen, Medical and Health Science Center, Institute of Cardiology
Móricz Zsigmond krt. 22. H-4032 Debrecen, Hungary
andras.durko@gmail.com
Background. Ischemic heart disease is a leading cause of death among women
worldwide, responsible approximately for one third of all cause mortality. In the
developed countries more women than men die because of IHD. Especially young
women (below age of 55 years) are at high risk, since compared to men at similar age
they are characterized by a worse outcome.
Objectives. The aim of the present study was to identify possible gender specific
cardiovascular risk factors in patients with ischemic heart disease. The study population
consisted of patients hospitalized with a diagnosis of ischemic heart disease, undergoing
diagnostic coronary angiography at the Department of Cardiology at the University of
Debrecen, MHSC in January 2007. The presence of regular cardiovascular risk factors
(age, positive family history, hypertension, diabetes mellitus, body weight,
hyperlipidemia, smoking) was documented. Relevant laboratory parameters on admission
(blood sugar level, glomerular filtration rate (GFR), total cholesterol, LDL-C, HDL-C,
triglyceride, hemoglobin (Hob)) were also analyzed and compared between women and
men patients.
Results. Average age of women and men patients was comparable (60.6±0.9 vs. 58.7±0.7
years, p=0.12), additionally in both patient groups one third of the patients was younger
than 55 years. Based on the documented risk factors more women than men suffered
from hypertension (82% vs.60%), type 2 diabetes mellitus (32% vs.17%) and were obese
(43% vs.26%). However, about twice as many smokers were observed among men than
women patients (18% vs.10%). Analysis of laboratory parameters revealed a significantly
higher HDL-C (1.59±0.05 vs. 1.32±0.03 moll/l, p<0.0001), but a significantly lower GFR
(89.4±1.3 vs. 96.4±1.4 ml/min, p=0.0038) and Hob (131±1 vs. 144±1 g/l, p<0.0001)
levels in women. Interestingly, GFR showed a significant decline with age in women
(r=0.66, p<0.001), but not in men patients (r=0.0016, p=0.98).
Conclusions. Based on the preliminary results of our study differences can be observed
between the distribution of cardiovascular risk factors in women and men patients with
ischemic heart disease. Our study also emphasizes the crucial importance of the careful
patient documentation in the clinical care. This data acquisition was associated with a
retrospective study initiated by the Regional Cooperation for Health, Science and
Technology (RECOOP HST) Consortium’s Women's Health and Cardiovascular
Diseases in Central and Eastern Europe Network.
66
Genetic effects on lung function and their associations with arterial
stiffness. Assumption or real link?
1Tarnoki D. L., 1Tarnoki A. D., 2Lazar Z., 3Cotichini R., 3Medda E., 3Fagnani C.,
3Stazi M. A., 3Nisticó L., 4Lucatelli P., 4Boatta E., 4Zini Ch., 4Fanelli F ,
5Baracchini C., 5Meneghetti G., 6Osztovits J., 6Jermendy G., 7,8Préda I., 7,8Kiss R.
G., 1Karlinge1 K., 9Lannert A., 7,8Molnar A. A.,, 10Littvay L., 11Garami Z.,
1Berczi V., 2Horvath I.
1 Department of Radiology and Oncotherapy, Semmelweis University, Budapest, Hungary
2 Department of Pulmonology, Semmelweis University, Budapest, Hungary
3 Istituto Superiore Di Sanitá, Italian Twin Registry, Rome, Italy
4 Department of Radiological Sciences, La Sapienza University of Rome, Rome, Italy
5 Department of Neurosciences, School of Medicine, University of Padua, Padua, Italy
6 Bajcsy Zsilinszky Hospital, III. Semmelweis University, Budapest, Hungary
7 Research Group for Inflammation Biology and Immunogenomics of Hungarian Academy of
Sciences and Semmelweis University, Budapest, Hungary
8 Department of Cardiology, State Health Center, Budapest, Hungary
9 Semmelweis University, School of Pharmacy, Budapest, Hungary
10 Central European University, Budapest, Hungary
11 The Methodist Hospital, DeBakey Heart and Vascular Center, Houston, TX, USA
tarnoki4@gmail.com
Objective: Central blood pressure (SBPao), peripherial and aortic pulse pressure (PP,
PPao), the difference between systolic and diastolic blood pressure in the ascending aorta
have consistently been shown to be a more powerful predictor of cardiovascular events
than traditional cuff blood pressure measurements taken at the arm. Twin studies by
comparing identical with non-identical twins produce information on the relative
contribution of genes and environment, and how the two interact. Purpose: To estimate
heritability and environmental effects on SBPao, PP and PPao and their correlations with
arterial stiffness (augmentation index on brachial artery, Aixbra, augmentation index on
aorta, Aixao, Pulse Wave Velocity on aorta, PWVao) using a twin sample. Methods and
Materials: 230 monozygotic (MZ) and 159 dizygotic (DZ) (Italian, Hungarian and
American) twin pairs were included in the study as part of International twin study 2009.
TensioMed Arteriograph was used to measure all parameters. Aix, SBPao and PWVao,
measured oscillometrically, showed strong correlation with the invasively obtained
values1. Statistical analysis was conducted using MPlus Version6. Results: Age, sex and
country-adjusted heritability of SBPao, PP and PPao indicated 45.5% (95% CI, 10.5 to
60.0%), 46.6% (95% CI, 29.8 to 58.0%), and 39.9% (95% CI, 1.4 to 53.9%). Unshared
environmental effects accounted for the largest part of variance, respectively (Table 1).
Model fit was normal. Bivariate saturated model showed high and significant correlations
between SBPao, PPao and arterial stiffness measures. Non-significant correlations were
estimated for PP and Aix (Table 2).
67
Table 1. Parameter Estimates and 95% CIs of the Best-Fitting Univariate Models
Measure h2 95% CI c2 95% CI e2 95% CI Model fit
(p)
SBPao,
mm Hg
0.455 0.105-
0.600
0.078 0.000-
0.388
0.467 0.382-
0.546
0.1464
PP, mm
Hg
0.466 0.298-
0.580
0.000 0.000-
0.027
0.534 0.432-
0.625
0.2569
PPao, mm
Hg
0.399 0.014-
0.539
0.050 0.000-
0.385
0.551 0.447-
0.661
0.4399
h2 indicates heritability; c2, shared environmental variance component; e2, unique environmental
variance component; and Model fit2, Chi-square test of Model fit (p value)
Table 2. Correlation (r) parameters and significance (p) values
Correlation values Aixbra, % Aixao, % PWVao, m/s
SBPao, mm Hg 0.588 (p<0.001) 0.587 (p<0.001) 0.475 (p<0.001)
PP, mm Hg -0.077 (p=0.057) -0.078 (p=0.055) 0.083 (p<0.05)
PPao, mm Hg 0.582 (p<0.001) 0.581 (p<0.001) 0.456 (p<0.001)
Conclusions: SBPao, PP and PPao are moderately heritable. Unshared environmental
effects account for the largest portion of the variance. High significant correlations were
estimated between atherosclerosis (arterial stiffness) and SBPao and PPao suggesting a
genetic background of these correlations. The measurement of central blood pressure and
aortic pulse pressure may be the next important advancement in the management of
hypertension and increased arterial stiffness.
The research was supported by Medexpert Ltd, Twins Days Festival Committee, Balassa
Institute Hungarian Scholarship Board Office, Ministry for Foreign Affairs of Republic of
Italy.
1. Horváth IG, Németh A, Lenkey Z, Alessandri N, Tufano F, Kis P, Gaszner B, Cziráki A.
Invasive validation of a new oscillometric device (Arteriograph) for measuring augmentation
index, central blood pressure and aortic pulse wave velocity. J Hypertens. 2010;28(10):2068-75.
68
The analysis of hepatic transcriptome in Prague Hereditary
Hypercholesterolemic (PHHC) rat – an experimental model of
hypercholesterolemia
1,2Zimolová M, 1Heczková M, 1,2Schmiedtová M, 1,2Poledne R, 1Jirsa M, 1,2Kovář J
1 Institute for Clinical and Experimental Medicine, Prague, Czech Republic
Videnska 1958/9, Prague, 14021Czech Republic
2 Centre of Cardiovascular Research, Prague, Czech Republic
mizi@ikem.cz
Most of the hypercholesterolemic patients have hypercholesterolemia of polygenic origin
and genes involved have not been well characterized yet. Unfortunately, there are almost
no animal models that could be used for study of polygenic hypercholesterolemia. Prague
Hereditary Hypercholesterolemic (PHHC) rat model may be an exception – PHHC rats
develop polygenic hypercholesterolemia after dietary cholesterol.
Objectives: To characterize pathogenesis of hypercholesterolemia and identify genes
involved in development of hypercholesterolemia in PHHC rat.
Methods: Firstly, nascent VLDL of PHHC and Wistar rats were isolated from serum
obtained 2 hours after intravenous application of tyloxapol and characterized. In another
experiment, serum VLDL was isolated from both PHHC and Wistar rats, labeled with 125I
and the rate of VLDL disappearance was determined after their application into Wistar
rats. Secondly, male PHHC and parental Wistar rats were fed chow (C) or 1% cholesterol
(CHOL) diet for three weeks. Hepatic transcriptome analysis was evaluated using
Affymetrix GeneChip arrays. The gene expression of selected genes was validated by
qPCR.
Results: On CHOL diet, cholesterol and triglycerides accumulated in the liver of both
PHHC and Wistar rats; cholesterolemia rose significantly only in PHHC rats. Nascent
VLDL of PHHC rats carry twice as much cholesterol than VLDL of Wistar rats on
CHOL diet and VLDL of PHHC rats are catabolized more slowly than those of Wistar
rats when injected in vivo. Nevertheless, the gene expression of both strains responded
to CHOL diet exactly in the same way - genes of cholesterol synthetic pathway were
down regulated. On the other hand, several genes were found to be differently expressed
between both strains independently of the diet (Aldh1a7, Yc2, Apof, Ugt2b, Cdh17, Ltc4s,
etc.). Apof as the only gene directly involved in lipoprotein metabolism was then
sequenced and the homozygous 17bp insertion was identified in the coding region in all
PHHC rats. This insertion was found also in Wistar rats with allelic frequency only 12 %.
Conclusions: PHHC rats secrete cholesterol-enriched VLDL from the liver and these
VLDL are catabolized more slowly. This can explain the accumulation of cholesterol in
serum of PHHC rats. The analysis of transcriptome revealed several new candidate genes
for hypercholesterolemia in our experimental model. Their exact role in pathogenesis
remains to be determined.
Supported by 1M0510 from MEYS CR.
69
Cardiovascular Research Network
Meeting Oral Presentation
70
Genetic Determinants of Cardiovascular Disease and its Risk
Factors
Gogalova V, Smolkova B, Dusinska M, Gasparovic J, Raslova K, Horska A,
Dzupinkova Z, Staruchova M, Volkovova K.
Slovak Medical University, Department of Experimental and Applied Genetics, Bratislava
Limbova 14, 83303 Bratislava, Slovakia
verona.gogalova@szu.sk
Background: Cardiovascular diseases (CVD) are the leading cause of death among non-
communicable diseases. A cross-sectional study of 2403 randomly selected subjects was
conducted to assess the relationship between several allelic variants of the genes
previously associated with the disease risk and CVD related quantitative traits.
Objectives: Variants of the Apo lipoprotein E (ApoE; rs429358 and rs7412),
methylenetetrahydrofolate reductase (NAD(P)H) (MTHFR; rs1801133),
methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 1-like (MTHFD1L;
rs6922269), chromosome 9p21.3 polymorphism (rs1333049), ATG16 autophagy related
16-like 1 (ATG16L1; rs2241880) and ATP-binding cassette, sub-family G (WHITE),
member 5 (ABCG8; rs11887534) were analyzed in relation to CVD related quantitative
risk factors. Studied variables were: body mass index, waist circumference, blood
pressure, serum total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C),
triglycerides (Tg), high density lipoprotein-cholesterol (HDL-C), Non-HDL cholesterol
(non HDL-C), Total to HDL cholesterol ratio (TC/HDL) and Glucose levels. Together
with positive personal and family anamnesis (case history) they were used for
determination of the classic CVD risk factors: obesity, dyslipidemia, hypertension,
diabetes, metabolic syndrome, smoking habit, and positive personal and family history.
Results: Variant rs429358 of the ApoE gene was strongly associated with elevated serum
lipid levels (TC P=0.000; LDL cholesterol P=0.000; triglycerides P=0.007), lowered
HDL-C levels (P=0.02) and higher nonHDL-C (P=0.000) and TC/ HDL-C ratio
(P=0.000). Rs7412 variant of the ApoE gene showed opposite effect on the serum lipids
(TC and LDL-C levels (P=0.000), HDL-C (P=0-048) also on nonHDL-C (P=0.000) and
TC/ HDL-C ratio (P=0.000). Higher frequency of variant allele rs429358 (P=0.001) and
lower frequency of the rs7412 (P=0.000) of the ApoE gene was found in the group of
dyslipidemic subjects. Variant rs2241880 of the ATG16L1 gene was associated with
higher BMI values (P=0.035) and waist circumference (0.02) in subjects with wild type
allele, who had lowest HDL-C and higher TC/HDL ratio (P<0.001). Similar results were
found for carriers of the rs11887534 variant of the ABCG8 gene. Wild type homozygotes
had elevated LDL-C (P=0.038) as well as both non HDL-C (P=0.014) and TC/HDL ratio
(P<0.009). None of the other variants was associated at p< 0.05 with the quantitative
traits tested here.
Conclusion: To summarize, we have identified four variants: rs429358 and rs7412 of
the ApoE, rs2241880 of the ATG16L1 and rs11887534 variant of the ABCG8 gene that
may affect risk of CVD and/or associate with quantitative CVD-risk factors.
71
Age related changes in rat cardiac spinal afferent neurons
Guić Maja Marinović, Runtić Branka, Grković Ivica
University of Split, School of Medicine in Split, Split, Croatia
Šoltanska 2, 21000 Split, Croatia
maja.marinovic.guic@mefst.hr
Background: Dorsal root ganglion (DRG) is a structure that contains ~90% of somatic
afferent neurons and only 5-10% of visceral afferent neurons. Within visceral afferent
neurons is a subpopulation responsible for the heart innervations, known as cardiac spinal
afferent neurons. Like many other cell types, the neurons constituting DRG are
susceptible to changes related to ageing. Previous studies in rat have shown that the
process of ageing does not affect the whole population of DRG neurons equally. In
addition, there was no change in proportion of isolectin B4 (IB4)-positive (likely
unmyelinated) and neurofilament 200 (N52)-positive (likely myelinated) neurons, while
the total number of neurons is slightly decreased or remains unchanged. However, to the
best of our knowledge, there are no studies about age-related changes of cardiac spinal
afferent neurons.
Objectives: The aim of this study was to investigate the effect of ageing on cardiac spinal
afferent neurons in the rat.
Materials and Methods: A patch loaded with retrograde tracer Fast Blue (FB) was
applied to all chambers of the rat heart. Morphological and neurochemical characteristics
of labeled cardiac spinal afferent neurons were assessed in young (2 months) and old (2
years) rats using IB4 and N52.
Results: Our study shows that the number of cardiac spinal afferent neurons decreased
with age and it was reduced to 15% of the total number of neurons found in young rats
(1604 vs. 248). In addition, we have observed a difference in myelinated and
unmyelinated neurons rate. The number of IB4-positive neurons increased significantly,
whereas the proportion of N52-positive ones decreased significantly during the ageing
process. The size of neuronal soma of IB4-positive neurons increased, while the size of
N52-positive neurons remained unchanged.
Conclusion: Unlike somatic spinal afferents, cardiac spinal afferent neurons underwent
morphological and neurochemical changes during ageing process. Although there is a
major decrease in total number of cardiac spinal afferent neurons, the innervation density
and nociceptive capacity are preserved due to both, an increased proportion and
compensatory growth of IB4-positive neurons.
72
Gender specifics of off-pump coronary bypass grafting
Rudenko M. L.
Institute of Cardiovascular Surgery of M.M Amosov
6 Mikola Amosov Str. Kyiv, Ukraine, 03680
n‐rudenko@yandex.ru
The aim of this work was to examine and compare the effect of gender on beating heart
CABG operations results, using compression type stabilizer.
Materials and Methods. In this study we used the database of the National
M.M.Amosov Institute of Cardiovascular Surgery. The study population consisted of
5113 patients undergoing isolated CABG between 2000 and 2009; 631 were female and
4482 were male. The patients were compared by gender, age, height, weight, ejection
fraction, amount of vessel disease and the usage of IMA. Amongst the 5113 surgeries
conducted, 4899 (95.8%) are done by the off-pump technique using compression type
stabilizer and only 214 (4,2%) are done by the on-pump technology mainly due to the
severity of the patients state. Amongst the on-pump CABG surgery patients, 87.1% were
men and 12.9% were women.
Results. Women tend to have more problems due to age, stature and different vessel
effect. Stature and age tend to be the worse predictors for CABG. The comparison of
male and female patients undergoing CABG shows that there were certain differences in
male and female patients. Women were older, of shorter stature, had higher ejection
fraction and had less one-vessel diseases. The most frequently grafted artery for both
groups (nearly 31 percent for both sex at all age) was the anterior interventricular artery.
In women on early stages (40-60 years old) the distal anastomosis is more frequently
conducted on artery marginalis (25.5% of all distal anastomosis), but in late stage (more
then 70 years old) in women the distal anastomosis is more frequently conducted on the
posterior interventricular artery (19.6% of all distal anastomosis). While the distal
anastomosis is more frequently conducted on artery marginalis (24.5% of all distal
anastomosis) for men in late stages (more then 70 years old), mortality between 2000-
2009 was 0.6% for men and 0.63% for women. Regarding off-pump technique, there was
no difference in mortality amongst patients off different sex. The main causes of death
are preoperative myocardial infarction and stroke.
Conclusion. The usage of beating heart technology using compression type stabilizer
allowed us to eliminate the influence of gender on coronary artery bypass grafting results.
73
Cardiovascular Research
Young Scientists’ Poster
Presentation
74
SCLO1B1 transporter and statin treatment efficacy
1,2Dlouhá D., 1Adámková V., 3Šnejderlová M., 3Češka R., 3Vrablík M.,
1,2Hubáček J. A.
1Institut for Clinical and Experimental Medicine;
2Cardiovascular Research Centre;
33rd Department of Internal Medicine, 1st Faculty of Medicine, Charles University;
Prague, Czech Republic
dadl@medicon.cz
Introduction: Statins belong to drugs of first choice in patients with increased
cardiovascular risk. There exists a significant inter-individual variability in statin
treatment efficacy that is likely to have a strong genetic background. Gene for SCLO1B1
belongs to the candidates with potential to influence the statin treatment efficacy.
SCLO1B1 codes for the solute carrier organic anion transporter (which has been shown
to regulate the hepatic uptake of statins and some other drugs).
Materials and methods: SCLO1B1 rs4149056 (T>C) polymorphism was analyzed on
heterogeneous group of 310 (141 males) patients with dyslipidemia treated by statins
(~90% on simvastin or atorvastatin, 10 or 20 mg per day). Polymorphism was analyzed
using PCR and restriction analysis. Lipid values (total-, LDL- and HDL- cholesterol,
triglycerides) were analyzed before the treatment and after 8 - 12 weeks of treatment.
Results: After treatment, there was a significant decrease both in total (7.4±1.3 →
5.4±1.0 mmol/L, P < 0.0001) and LDL-cholesterol (4.7±1.2 → 3.2±0.9 mmol/L, P <
0.0005). Distribution of the individual genotypes in Czech patients (TT = 62%, CT =
32%, CC = 6%) was similar to the distributions found previously in west European
populations. Decrease of both total cholesterol (29% vs. 24%) and LDL cholesterol (34%
vs. 30%) was non - significantly higher in the carriers of the CC genotype in comparison
with the commonest TT homozygotes. Changes in values of the HDL - cholesterol and
triglycerides were not associated with analyzed polymorphism.
Conclusions: Results of the pilot study suggests that the rs4149056 variant within the
gene for SCLO1B1 transporter could be one of the genetic determinants of statin
treatment efficacy in Czech patients with dyslipidemia. Because of the low frequency of
the potentially advantageous genotype, it is necessary to analyze larger group of patients
Supported by project No. NS 11-307 (IGA MH CR).
75
Adrenergic receptors upregulation in ovariectomized rats
1Viljetić Barbara, 2Blažeković Robert, 3Perkov Vedrana, 3Heffer Marija
1 Department of Chemistry, Biochemistry and Clinical Chemistry, School of Medicine, Josip Juraj
Strossmayer University of Osijek, Osijek, Croatia
2 Department of Cardiac Surgery, University Hospital Center Osijek, Osijek, Croatia
3 Department of Medical Biology, School of Medicine, Josip Juraj Strossmayer University of Osijek,
J.Huttlera 4 31000 Osijek, Croatia
bviljetic@mefos.hr
Stress is gender specific response aimed to preserve physiological homeostasis. Although
it is whole organism reaction, the stimulation originates from two sides: hypothalamic-
pituitary-adrenal axis (HPA) and sympathetic-adrenal medullary system. Both sides,
either through norepinephrine or epinephrine, converge on Beta-1 adrenergic receptor,
which is G-protein coupled membrane receptor. Activation of the heart Beta-1 receptors
cause increased cardiac output by rising heart rate, increasing impulse conduction and
ejection fraction – all required in stress response, but inadequate in the state without real
jeopardy.
Three-month-old rats were ovariectomized and kept for a month in the unstressful
environment. Animals were sacrificed and compared with age-matched male and female
controls. Sampled heart, hypothalamic and fat tissue were immunoprobed for Beta-1 and
Beta-3 receptors. The heart tissue of ovariectomized animals had highest concentration of
Beta-1 receptors.
The Beta-1 receptor upregulation in a condition of ovarian hormone deprivation could
have serious consequence on cardiac function, particularly in stress response. We assume
that heart becomes more sensitive to levels of circulating norepinephrine and
epinephrine, overreacting even on mild provocation. Similar changes might be
pathophysiological mechanism in the cardiovascular disease in human premature ovarian
failure and polycystic ovary syndrome.
76
High salt diet is associated with impaired microvascular reactivity
in young healthy female human subjects
1*Grizelj Ivana , 1*Čavka Ana , 2Jelaković Bojan, 3Lombard Julian H, 4Mihaljević
Ivan, 5Koller Akos, 1Drenjančević Ines
1Department of Physiology and Immunology, School of Medicine J.J. Strossmayer University of
Osijek, Osijek, Croatia
2Department of Internal Medicine, Division of Nephrology and Arterial Hypertension, University
Hospital Center Zagreb, Zagreb, Croatia
3 Department of Physiology, Medical College of Wisconsin, Milwaukee, USA
4 Clinical Institute of Nuclear Medicine and Radiation Protection, Clinical Hospital Centre
Osijek,Osijek, Croatia
5Dept of Patophysiology and gerontology, University of Pecs, Pecs, Hungary
*I.Grizelj and A.Čavka equally contributed to the abstract
grizelj.ivana@gmail.com
Studies on experimental animals have shown that changes in salt intake significantly alter
vascular reactivity to different physiological stimuli, in conduit vessels, resistance
arteries, and in the microcirculation. However, the effects of high salt (HS) intake on
microvascular endothelial response in young healthy human subjects without pre-existing
conditions such as diabetes and hypertension are still unknown.
The aim of this study was to assess effects of acute salt loading on microvascular
reactivity during reactive hyperemia in young healthy women, using non-invasive Laser
Doppler Flowmetry (LDF). Eleven normotensive women (21±3 years) were instructed to
maintain a low-salt (LS) diet (less than 40 mmol Na/daily) during 7 days. Simultaneously
they were divided into HS group (N=6) (intake of 200 mmol Na/daily) and placebo group
(N=5). LDF was performed before and after salt diet protocol as measurement of relative
changes in blood flow between baseline flow and reperfusion that follows 1- and 2-
minute occlusion. Blood sampling for plasma electrolytes, aldosterone and plasma renin
activity, just as 24-hour urine sodium and potassium excretion took place before and after
diet protocol.
In the HS group there was a statistically significant decrease in micro vascular reactivity
after 1-minute occlusion (endothelium-dependant mechanisms) with no difference after a
2-minute occlusion (maximum dilation ability), before and after HS diet. The increased
urinary volume, decreased urinary sodium, and increased urinary potassium
concentration in subjects on LS diet, and increased urinary sodium concentration in
subjects on HS diet confirmed consistency of experimental protocol and subjects’
adherence to diet. Although plasma renin activity and serum aldosterone concentration
decreased in HS diet and increased in women on LS diet, they didn’t reach statistical
significance.
This study shows that even one week of HS intake may have negative effect on micro
vascular reactivity, decreasing blood flow during reactive hyperemia by affecting
endothelial function. Increased number of subjects in further study is needed.
77
Antioxidant status in the context of obesity in humans.
Horska Alexandra, Staruchova Marta, Mislanova Csilla, Spustova Viera,
Volkovova Katarina
Department of Experimental and Applied Genetics, Slovak Medical University,
12, Limbova, 833 03 Bratislava, Slovakia
alexandra.horska@szu.sk
Background: Obesity is multifactorial chronic disorder defined as abnormal, excessive
body fat accumulation, leading to increased health problems, reduced life expectancy and
discrimination. Genetic predisposition, metabolic, hormonal and mostly behavioral
aspects are implied in the pathogenesis of this phenomenon, which is regarded to be
global epidemic of recent time. Overweight and obesity represent major risk factors for
chronic diseases such as type 2 diabetes, hypertension, cardiovascular diseases,
osteoarthritis and certain cancers. Increased systemic oxidative stress in accumulated
adipose tissue is one of pathogenic mechanisms of obesity-associated metabolic
syndrome and cardiovascular diseases.
Objectives: For our purposes, 97 normal weight, 95 overweight and 94 subjects with
BMI>30 were selected from cross-section study of 40 years old probands from the
Slovak population. Spectrum of biochemical parameters, lipid profile, antioxidant status
and other markers were analyzed in blood of all volunteers. Polymorphisms in 27
candidate genes involved in etiology of common chronic diseases and in genes for
defense mechanisms were determined to assess individual genetic susceptibility.
The aim of this study was to investigate markers of antioxidant defense and oxidative
stress, and seek for associations between genetic background and clinical, biochemical
parameters in obese population.
Results: In 73% of overweight and 90% of obese subjects, systolic and diastolic blood
pressure values were above recommended reference 120/80 mmHg. Individuals with
BMI>30 showed increased activity of ceruloplasmin oxidase (p<0.05) and decreased
level of cysteine (p<0.05). Reduced glutathione was decreased in overweight subjects too
(p<0.05). Concentrations of retinol (p<0.05) and tocopherols (p<0.05) in plasma were
elevated when BMI>25. Reversely, levels of xanthophylls (p<0.01) and β-carotene
(p<0.001) were decreased in subjects with BMI>30. In all overweight and obese subjects
was observed markedly decreased protection of fatty acids against peroxidation,
calculated from vitamin E/triacylglycerol´s ratio (p<0.001).
Subjects with deletion of detoxification and antioxidant enzyme GSTT1 displayed lower
activity of glutathione peroxidase in erythrocytes, compared with subject owning
efficient enzyme (p<0.05).
Conclusions: Several parameters of antioxidant state were found changed in overweight
and obese subjects, which indicate elevation of oxidative stress in these groups. Redox
state is an early marker and could serve as potential therapeutic target for obesity-related
cardiovascular disorders.
Work was supported by the Ministry of Health of SR project No. MZ 2006/07-SZU-02.
78
Adenosine triphosphate concentration of exhaled breath condensate
in asthma
Lázár Zsófia, Orosz Márta, Gálffy Gabriella, Bikov András, Losonczy György,
Horváth Ildikó
Department of Pulmonology, Semmelweis University, Budapest, Hungary
1/C Diosarok Str. 1125 – Budapest Hungary
zsofia.lazar@yahoo.com
The pathogenesis of bronchial asthma, a common obstructive inflammatory airway
disease affecting 300 million people worldwide, is not clearly understood. Adenosine
triphosphate (ATP), a mediator of purinergic signaling, might play a role in disease
development by triggering airway inflammation and inducing bronchoconstriction.
However, supporting human evidence is scarce. ATP can be measured in exhaled breath
condensate (EBC), a non-invasive airway sample generally suitable for disease
monitoring. For the first time, we aimed to determine EBC ATP concentration in asthma,
and to study its relation to disease parameters and calculate airway ATP level.
EBC was collected from 45 patients with persistent asthma (age 34.7 (yr) ± 13.2; FEV1 %
predicted 87.0 ± 15.5; mean ± SD) and 32 healthy controls (age 36.9 ± 12.6; FEV1 %
predicted 98.9 ± 9.9). Exhaled nitric oxide and spirometry were measured; disease
control was assessed with the Asthma Control Test. ATP was measured in luciferin-
luciferase assay. Airway ATP concentration was calculated using dilution estimated from
conductivity of vacuum-treated EBC samples. Parametric tests were applied; ATP
concentrations and nitric oxide levels were logarithmically transformed (geometric mean
[95% confidence interval]).
EBC ATP concentrations were similar in asthmatic patients and healthy controls (4.12
pM [3.13-5.41]
vs. 3.18 pM [2.32-4.36], p=0.21) and it was not related to lung function variables,
exhaled nitric oxide level or disease control. EBC ATP concentration was correlated to
airway droplet dilution in the sample (r=–0.32, p<0.05). The calculated airway ATP level
was not elevated in patients compared to controls (24.79 nM [17.27-35.58] vs. 20.70 nM
[13.04-32.86], p=0.52), but it was negatively related to FEV1 in study subjects (r=–0.35,
p<0.05). The estimated airway ATP concentration was similar to that previously
measured in airway epithelial cultures.
EBC ATP concentration might not serve as a biomarker of bronchial asthma.
Nevertheless our data provide the first evidence that the assessment of respiratory droplet
dilution in EBC can be reliably applied to calculate real mediator level in the airways
thereby increasing the applicability of EBC technique for monitoring of the airways. Our
results also demonstrate that extracellular ATP might be involved in a general
mechanism regulating airway caliber.
79
Trend of change in gender distribution among patients with acute
coronary syndrome in Slavonia and Baranja region (Croatia)
1Makarović Zorin, 1Makarović Sandra, 2Mlinarević Dražen, 2Lulić Dinka, 1Steiner
Robert
1 Department of Cardiology, University Hospital Center Osijek, Osijek, Croatia
2 Department of Medical Biology, School of Medicine, Josip Juraj Strossmayer University of Osijek,
Osijek, Croatia
Huttlerova 4, 31 000 Osijek, Croatia
zorinmakarovic2@net.hr
First catheterization laboratory in Osijek region was opened in 2005 and was followed by
institution of cardiosurgery during 2007. The laboratory had managed 88 myocardial
infarctions with ST elevation (STEMI) by primary percutaneous coronary intervention
(PCI) before the cardiosurgical treatment was available. With the surgeon in site the
number of the patients has rapidly raised up to 106 primary PCIs in just two months.
Most of the patients with STEMI managed with primary PCI (PPCI) before 2007 (70.7%)
and during 2008 were males (80.6 %). Mean age of the male patients was 55.2 before
2007 and 58.1 years in 2008, while the mean age of the female patients was 65.8 and 59.9
years, respectively.
Looking closely the patients undergoing coronarography (48 in total) during January
2007 we found that 54% of them were males (mean age 57.5 years). Females’ mean age
was 59.1 years. Long lesions (≥20 mm) were found in 2 males and in 1 female patient.
Interestingly, the lesions in small vessels were more frequently found in females (n=13)
than in males (n=7). There were 38 coronary stents placed during the PCI (36 bare metal
stents and 2 drug-eluting stents). There was only one female patient without significant
coronary artery disease. Three female patients experienced artificially caused coronary
artery dissection. The outcome of the coronarography was coronary artery bypass graft
(CABG) surgery in 9 males and in 4 female patients. One male and one female patient
underwent urgent CABG surgery. Equal number of male and female patients was treated
with optimal medical therapy (4).
The total number of the patients managed by coronarography in 2007 was 492. Thereof
were 182 PCI cases (102 for ongoing infarction, 69 during the diagnostic study and 3
with multivessel PCI during the one session) and 237 stents placements.
In 2009 and 2010 the catheterization laboratory managed 906 and 1056 patients,
respectively, out of which 60 % were males. In 30 % of patients the diagnostic procedure
was followed by PCI and percutaneous transluminal coronary angioplasty. Nevertheless,
the number of urgent PCIs rose up to 96 in 2009. We observed a trend of rising number
of female patients, accompanied with younger age and pathology of small vessels. This
phenomenon is contrary to the predominant clinical expectations and suggests some
drastic changes in gender specific epidemiology of coronary disease.
80
Metabolic background of heart rate variability: Evidence achieved
by supplementation with amaranth oil in type 2 diabetes mellitus
patients
Semen Khrystyna, Kaminskyy Danylo, Yelisyeyeva Olha
Danylo Halytsky Lviv National Medical University, Lviv, Ukraine
69, Pekarska str., 79010, Lviv, Ukraine
khrystyna_semen@yahoo.com
Heart rate variability (HRV) is a modern noninvasive tool frequently used in clinical practice
to establish functional capacity and prognosis in patients with various illnesses, particularly
diabetes mellitus (DM). This disorder is associated with significant reduction of HRV
indexes and oxidative stress (OS) related to excessive accumulation of oxidative destruction
products with depression of redox reactions in cells and tissues. There is increasing evidence
about close relationships between HRV and certain metabolic parameters reflecting the depth
of OS, but biochemical mechanisms of these relationships remain to be established. The aim
of present research was to study the changes of HRV and some parameters of aerobic
metabolism in type 2 DM patients achieved with supplementation with oil derived from the
seeds of Amaranthus cruentus L. (AmO).
The impact of concentrated AmO (1.0 ml per 60 kg of body weight daily for one month) on
the time and frequency domain HRV indexes and some parameters reflecting OS in blood
(levels of TBARS, oxidative modification proteins (OMP), hydro peroxides (HP), LDL-
cholesterol, middle mass molecules (MMM), activities of catalase and superoxide dismutase
(SOD)) were evaluated in 36 patients with moderate and severe type 2 DM aged 51-67 years
with disease duration up to 10 year. Control group consisted of 35 apparently healthy
volunteers of the same age. Severe manifestations of OS were found in all DM patients
despite increased activities of catalase and SOD. These biochemical changes were associated
with dramatically decreased HRV. Because of wide range of total power (TP) values
obtained, the patients were divided into 2 groups, with group 1 including the subjects with
low (600-1000ms2) and group 2 very low (100-600ms2) TP indexes. Administration of AmO
caused activation of aerobic metabolism with significant decrease in TBARS, HP, LDL, and
OMP levels in group 1 patients. Simultaneously, significant increase in most HRV
parameters, predominantly due to autonomic up regulation was observed in this group. The
patients with initial low TP values (group 2) demonstrated normalization of antioxidant
enzymes activities, however, no changes were observed in patients with initially high levels
of TBARS, MMM, and OMP. Notably, marked decrease in HP, LDL was shown in group 2
due to better utilization of these OS markers in redox reactions, which was accompanied by
improvement of HRV parameters.
In conclusion, activation of aerobic metabolism with AmO supplementation in type 2 DM
patients caused more prominent increase in HRV indexes in group 1 (with initially higher
TP) patients. That was associated with the improved intensity of aerobic metabolism and
reduction of OS manifestations. Thus, combination of biochemical studies and HRV
provides more precise assessment of functional and metabolic conditions, more adequate
follow up, and control of treatment.
81
Genetic effects on lung function and their associations with arterial
stiffness. Assumption or real link?
1Tarnoki D. L., 1Tarnoki A. D., 2Lazar Z., 3Cotichini R., 3Medda E., 3Fagnani C.,
3Stazi M. A., 3Nisticó L., 4Lucatelli P., 4Boatta E., 4Zini Ch., 4Fanelli F ,
5Baracchini C., 5Meneghetti G., 6Osztovits J., 6Jermendy G., 7,8Préda I., 7,8Kiss R.
G., 1Karlinge1 K., 9Lannert A., 7,8Molnar A. A.,, 10Littvay L., 11Garami Z.,
1Berczi V., 2Horvath I.
1 Department of Radiology and Oncotherapy, Semmelweis University, Budapest, Hungary
2 Department of Pulmonology, Semmelweis University, Budapest, Hungary
3 Istituto Superiore Di Sanitá, Italian Twin Registry, Rome, Italy
4 Department of Radiological Sciences, La Sapienza University of Rome, Rome, Italy
5 Department of Neurosciences, School of Medicine, University of Padua, Padua, Italy
6 Bajcsy Zsilinszky Hospital, III. Semmelweis University, Budapest, Hungary
7 Research Group for Inflammation Biology and Immunogenomics of Hungarian Academy of
Sciences and Semmelweis University, Budapest, Hungary
8 Department of Cardiology, State Health Center, Budapest, Hungary
9 Semmelweis University, School of Pharmacy, Budapest, Hungary
10 Central European University, Budapest, Hungary
11 The Methodist Hospital, DeBakey Heart and Vascular Center, Houston, TX, USA
tarnoki4@gmail.com
Background: An association between reduced lung function and an increased risk of vascular
events has been reported but the underlying mechanisms are unknown. So far no twin studies
have examined the association between atherosclerosis and lung function. Objectives: To
estimate association of arterial stiffness and lung function and to assess heritability and
environmental effects on these parameters. Subjects and Methods: 380 (232 monozygotic /MZ/
and 148 dizygotic /DZ/) twin pairs (age yrs 50.5±15.4; mean±SD) were included in this classical
twin study as part of International Twin Study 2009. Augmentation index on brachial artery
(Aixbra) and Pulse Wave Velocity on aorta (PWVao) were measured to reflect arterial stiffness
(TensioMed Arteriograph) and forced vital capacity (FVC) with forced expiratory volume in one
second (FEV1) (MIR Minispir) for lung function. Heritability and Pearson correlation coefficient
(β) between variables were determined. Results: Age and gender-corrected heritability of FVC
and FEV1 were 0.74 and 0.75 (p<0.01). Shared and unshared environmental effects were found
to be 0, 0.26 (p<0.01) and 0, 0.25 (p<0.01) for FVC and FEV1, respectively (Table 1). β between
FVC and PWVao were -0.42 (p<0.001) and -0.29 (p<0.001), respectively. β between FEV1 and
Aixbra and PWVao were found to be -0.47 (p<0.001) and -0.38 (p<0.001), respectively. Table 1.
Heritability (A), common (C) and unique (E) environmental effects of FVC and FEV1
Conclusions: Lung function is strongly heritable and is associated with arterial stiffness. The
observed relationship can aid to understand the background of vascular changes in different
airway diseases. The research was supported by Medexpert Ltd, OTKA 68808 grant,
Twins Days Festival Committee,
Balassa Institute Hungarian
Scholarship BoardOffice,
Ministry for Foreign Affairs of
Republic of Italy.
A C E
FVC 0.74 (p<0.01) 0 0.26 (p<0.01)
FVC %predicted 0.422 0.107 0.471 (p<0.01)
FEV1 0.747 (p<0.01) 0 0.253 (p<0.01)
FEV1 %predicted 0.002 0.003 0.995 (p<0.01)
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