Adiponectin resistance parameter as a marker for high normal blood pres-sure and hypertension in patients with metabolic syndrome.

INTRODUCTION: The different degrees of adiponectin/insulin sensitivity and dysfunctional adipose tissue lead to the development of hypertension (HT). This study aimed to determine adiponectin (AD) concentration in patients with metabolic syndrome (MetS) and high-normal blood pressure or hypertension and to investigate the importance of Homeostatic Model Assessment-AD (HOMA-AD) index in assessing adiponectin/insulin resistance in hypertension. METHODS: This cross-sectional study involved 150 subjects divided into two groups: with MetS (and high-normal blood pressure, n =50; and HT, n =50), and controls without MetS (n =50). In all subjects, serum adiponectin concentration was measured by enzyme-linked immunosorbent assay method. Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) and HOMA-AD index were calculated. RESULTS: The results showed that, compared to the control group, serum AD concentrations were significantly lower in patients with MetS and high-normal blood pressure (p =0.008), and the lowest in group MetS and HT (p =0.001). High AD levels and low HOMA-AD were significantly associated with decreased blood pressure values. In patients with MetS, the value of HOMA-AD≥1.13 was associated with a higher risk of developing high-normal blood pressure. Furthermore, the value of HOMA-AD≥2.63 was associated with a higher risk of developing hypertension. CONCLUSIONS: Hypoadiponectinemia is associated with hypertension, especially in the early stages of the disease. The serum AD levels and HOMA-AD index may be useful markers for identifying patients at risk for high-normal blood pressure and hypertension. HIPPOKRATIA 2020, 24(1): 3-7.

PPAR𝛾 Gene and Atherosclerosis: Genetic Polymorphisms, Epigenetics and Therapeutic Implications.

Atherosclerosis is the leading cause of mortality and morbidity in the developed world. It is characterized by the formation of a plaque in the walls of middle and large arteries leading to macrovascular complications. Several risk factors are included, with diabetes being one of the most important for the onset and development of atherosclerosis. Due to an increase in the prevalence of diabetes in the world, the incidence of diabetic complications (microvascular and macrovascular) is increasing. Peroxisome proliferator-activated receptor γ (PPARγ) plays a important role in atherosclerotic processes. Peroxisome proliferator activated receptor γ belongs to the superfamily of nuclear receptors, has a great presence in fat tissue, macrophages, and regulates gene expression and most of the processes that lead to the onset and development of atherosclerosis. In this review, we discuss the basic patho-physiological mechanisms of atherosclerosis in type 2 diabetes mellitus (T2DM). Furthermore, we discuss the impact of PPARγ polymorphisms, and the epigenetic mechanisms affecting the onset of atherosclerosis, i.e, DNA methylation and demethylation, histone acetylation and deacetylation, and RNA-based mechanisms. Moreover, we add therapeutic possibilities for acting on epigenetic mechanisms in order to prevent the onset and progression of atherosclerosis.

ADIPONECTIN AS A POTENTIAL BIOMARKER OF LOW BONE MINERAL DENSITY IN POSTMENOPAUSAL WOMEN WITH METABOLIC SYNDROME.

CONTEXT: Adiponectin is an abundant adipokine, which has antiinflammatory, anti-atherosclerotic and vasoprotective actions, and potential antiresorptive effects on bone metabolism. It seems to be directly involved in the improvement and control of energy homeostasis, protecting bone health and predicting osteoporotic fracture risk. OBJECTIVE: To examine the relationship between adiponectin level and bone mineral density (BMD) in post-menopausal women with metabolic syndrome (MetS) and low BMD, and to estimate the prognostic significance of adiponectin in osteoporosis. DESIGN: Clinical-laboratory cross-sectional study including 120 middle-aged and elder women (average 69.18±7.56 years). SUBJECTS AND METHODS: The anthropometric parameters were measured for all examinees. Lumbar spine and hip BMD, as well as body fat percentage, were measured using a Hologic DEXA scanner. In all subjects serum adiponectin concentration was measured by ELISA method. RESULTS: The level of adiponectin was significantly positively correlated with BMD-total, BMD of the lumbar spine and BMD of the femoral neck (r=0.618, r=0.521, r=0.567; p<0.01). Levels of adiponectin and BMD are significantly lower in post-menopausal women with MetS and osteoporosis compared to patients with osteopenia (856.87±453.43 vs. 1287.32±405.21 pg/mL, p<0.01; BMD, p<0.05), and the highest values in healthy examinees. A cut-off value of adiponectin level for osteoporosis/osteopenia was 1076.22/1392.74 pg/mL. CONCLUSIONS: Post-menopausal women with MetS have significantly lower adiponectin level and low BMD compared to healthy examinees. Adiponectin may be an early, significant and independent predictor of developing osteoporosis in women with MetS, especially in post-menopausal period.

PECAM-1 gene polymorphism (rs668) and subclinical markers of carotid atherosclerosis in patients with type 2 diabetes mellitus.

The platelet endothelial cell adhesion molecule 1 (PECAM-1) plays an important role in many inflammatory processes, including the development of atherosclerosis. Polymorphism rs668 of the PECAM-1 gene (373C/G) is functional, and it was reported to be associated with increased serum levels of PECAM-1. We investigated the association between the rs668 polymorphism of PECAM-1 and subclinical markers of carotid atherosclerosis in subjects with type 2 diabetes mellitus (T2DM). Five hundred and ninety-five T2DM subjects and 200 control subjects were enrolled. The carotid intima-media thickness (CIMT) and plaque characteristics (presence and structure) were assessed ultrasonographically. Biochemical analyses were performed using standard biochemical methods. Geno-typing of the PECAM-1 gene polymorphism (rs668) was performed using KASPar assays. The control examinations were performed 3.8 ± 0.5 years after the initial examination. Higher CIMT was found in patients with T2DM in comparison with subjects without T2DM. Statistically sig-nificantly faster progression of the atherosclerotic markers was shown in subjects with T2DM in comparison with the control group. When adjusted to other risk factors, the rs668 GG genotype was associated with an increased risk of carotid plaques in subjects with T2DM. We concluded that our study demonstrated a minor effect of the rs668 PECAM-1 on markers of carotid atherosclerosis in subjects with T2DM.

Mast Cells Might Have a Protective Role against the Development of Calcification and Hyalinisation in Severe Aortic Valve Stenosis.

Aortic valve stenosis is characterized by inflammation and extracellular matrix remodelling. The aim of this study was to analyse the impact of mast cells on the occurrence of histopathological changes of aortic valves in patients with severe grade, non-rheumatic degenerative aortic valve stenosis. Valve specimens were obtained from 38 patients undergoing valve replacement. The role of mast cells was analysed by dividing the specimens into two groups, characterized by the presence (group A, N = 13) or absence of mast cells (group B, N = 25). There were no significant differences in clinical data between the two groups. In group A, T cells and macrophages were present in all aortic valves, as compared to a significantly lower proportion of valves with T cells and macrophages in group B. Valves in group A were less often calcified and hyaline-degenerated than valves in group B. There were no changes in fibrosis between the two groups. We found a positive correlation between the presence of mast cells and macrophages/T cells, a negative correlation between the presence of mast cells and calcification/ hyaline degeneration, and no correlation between the presence of mast cells and fibrosis. There was also a negative correlation between the presence of macrophages/T cells and calcification. The linear regression model identified only the presence of mast cells as an independent negative prediction value for calcification. In conclusion, mast cells might have a protective role against the development of calcification and hyaline degeneration in severe grade, non-rheumatic aortic valve stenosis.

The impact of thyroid hormone replacement therapy on left ventricular diastolic function in patients with subclinical hypothyroidism.

OBJECTIVE: Subclinical hypothyroidism (SH) is associated with a moderately elevated risk of heart failure events among older adults. The objective of our prospective study was to assess the impact of thyroid hormone replacement therapy (HRT) with low doses of L-thyroxine (6.25-25 µg/day) on left ventricular diastolic function in patients with SH. MATERIALS AND METHODS: 33 patients with SH and 25 healthy controls were involved. All participants underwent standard echocardiography and Doppler imaging at baseline and, the patient group, also after a course of HRT. RESULTS: At baseline, patients with SH showed significantly lower E (0.79 ± 0.22 vs. 0.93 ± 0.19, p < 0.001), E/A ratio (1.19 ± 0.29 vs. 1.31 ± 0.25, p < 0.003), and higher intraventricular septum thickness (IVST) (0.99 ± 0.14 vs. 0.89 ± 0.18, p < 0.001) in comparison with healthy controls. After 6 months of therapy, the E/A ratio underwent significant increase (1.28 ± 0.21 vs. 1.19 ± 0.29, p < 0.001), while the IVS displayed a robust reduction (0.92 ± 0.16 vs. 0.99 ± 0.14, p < 0.001). CONCLUSIONS: HRT with low-dosed L-thyroxine may improve left ventricular diastolic function in patients with SH.

Polymorphism of angiotensin-converting enzyme (rs4340) and diabetic nephropathy in Caucasians with type 2 diabetes mellitus.

Diabetic nephropathy (DN) is the leading cause of endstage renal disease (ESRD) in developed countries. Several environmental and genetic factors predict the development and progression of DN. The renin-angiotensin system was demonstrated to be involved in the development of DN. We evaluated the association between rs4340 of the angiotensin-converting enzyme (ACE) gene and DN in Caucasians with type 2 diabetes mellitus (T2DM) in 276 Slovenian patients with T2DM who had DN, and 375 patients without clinical signs of DN. Genetic analysis was performed with either standard polymerase chain reaction (PCR) (for rs4340). Results were analyzed using the χ2 test and multivariate logistic regression analyses. We found no association between rs4340 and DN. Cystatin C was significantly higher in the DN+ group (p <0.001) than in the DN group. Cystatin C was a better marker for the estimation of renal function than estimated glomerular filtration rate (eGFR) according to the modification diet in renal disease (MDRD) equation mL/ min. We concluded that there was no association between the rs4340 of the ACE gene and DN in Caucasian patients who have T2DM.

Association of the ACE rs4646994 and rs4341 polymorphisms with the progression of carotid atherosclerosis in slovenian patients with type 2 diabetes mellitus.

The current study was designed to reveal possible associations between the angiotensin-converting-enzyme (ACE) gene polymorphisms (rs4646994 and rs4341) with markers of carotid atherosclerosis in patients with type 2 diabetes mellitus (T2DM) in a 4-year-long follow-up study. Five hundred and ninety-five T2DM subjects and 200 control subjects were enrolled. Genotyping of ACE polymorphisms was performed using KASPar assays, and ultrasound examinations were performed twice (at the enrollment and at follow-up). With regard to the progression of atherosclerosis in subjects with T2DM, statistically significant differences were demonstrated in the change of the sum of carotid plaques thickness for the rs4646994 polymorphism. We did not demonstrate an association between the tested polymorphisms (rs4646994 and rs4341) and either carotid intima media thickness (CIMT) or CIMT progression in a 3.8-year period. In our study, we demonstrated that subjects with T2DM with the DD genotype of the rs4646994 [ACE insertion/deletion (I/D)] polymorphism had faster progression of atherosclerosis in comparison to subjects with other genotypes.

Polymorphisms +45T>G and +276G>T of the adiponectin gene does not affect plasma adiponectin level and carotid intima-media thickness in patients with diabetes mellitus type 2.

AIM: Despite increasing evidence of adiponectin's anti-inflammatory and antiatherogenic effects, its role in atherogenesis remains uncertain. The aim of the present study is to investigate the association between +45T>G and +276G>T polymorphisms of the adiponectin gene and both plasma adiponectin levels and carotid intima-media thickness in patients with diabetes mellitus type 2. METHODS: 301 diabetic patients, divided into three categories on the basis on BMI were enrolled in the study. Carotid intima-media thickness (CIMT) was assessed ultrasonographically. Plasma adiponectin levels were measured by enzyme-linked immunosorbent assay (ELISA). Genotypes were determined by real-time PCR. RESULTS: Adiponectin level and prevalence of the G allele of 45T>G polymorphism decreased significantly with increasing BMI category. G allele of +45T>G polymorphism was associated with higher plasma adiponectin level only after adjustment for age, sex and BMI. No statistically significant difference in CIMT and +276T>G genotypes distribution was observed between BMI categories. None of the polymorphisms as well as plasma adiponectin level was associated with CIMT after adjustment for covariates. CONCLUSION: The G allele of the +45T>G polymorphism is not independently associated with plasma adiponectin level and is not associated with CIMT. +276G>T polymorphism is not associated with plasma adiponectin levels and CIMT in diabetic patients.

[Prosthetic rehabilitation in patient with forced class III malocclusion].

The paper presents clinical case of 63 years old edentulous patient with slight class III malocclusion. For 15 years he was using inadequately fabricated dentures causing forced severe class III malocclusion. Forced progeny was corrected by newly fabricated dentures which restored normal orofacial function and facial harmony.

The semi-sewer river: hydraulic backwater effects and combined sewer overflow reverse flows in Central Brussels reduce deoxygenation impact further downstream.

In 2011 and 2012 the dissolved oxygen content in the low-discharge river Zenne was monitored continuously, every 5 minutes, downstream of Brussels city centre, making it possible to document the complex mechanisms by which combined sewer overflow (CSO) spills affect both the hydraulics and the oxygen balance of the hydrosystem. In addition to oxygen demand impacts, proportions of water volumes are such that the oxygen-devoid sewage water discharged from CSOs contributes significantly to the oxygen deficit observed in the river further downstream. It is shown that ensuing unexpected hydraulic behaviour, such as a full river-flow reversal, can explain the dual nature of oxygen sag following major CSO events. At times the semi-sewer river plays the role of an in-stream stormwater tank, effectively attenuating the environmental impacts of Brussels CSOs.

Association of manganese superoxide dismutase and glutathione S-transferases genotypes with carotid atherosclerosis in patients with diabetes mellitus type 2.

AIM: The aim of the present study was to test the association between genetic polymorphisms with functional effects on redox regulation: Ala16Val of manganese superoxide dismutase (MnSOD or SOD2), polymorphic deletions of glutathione S-transferases M1 (GSTM1) and T1 (GSTT1) and Ile105Val of the GSTP1 and carotid atherosclerosis in patients with type 2 diabetes. METHODS: The study enrolled 287 subjects with type 2 diabetes. Carotid atherosclerosis was quantified by ultrasonography as carotid intima-media thickness (CITM), plaque score from 0 to 6 and plaque type from 1 to 5. Genotypes were determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). RESULTS: The highest triglyceride level was observed in patients with MnSOD Val/Val genotype. Other polymorphisms did not show significant association with clinical parameters. We did not observe significant differences in MnSOD, GSTM1 and GSTP1 genotypes distribution according to CIMT, plaque type or plaque score. After adjustment for age, sex, smoking, BMI, lipid parameters and duration of hypertension and diabetes carriers of GSTT1-0 genotype showed an increased risk for higher plaque score (OR=2.29; p=0.012), but no association with CIMT and plaque stability was observed. Carrying of both GSTM1-0 and GSTT1-0 did not influence clinical parameters but increased risk for higher plaque score (OR=2.59; P=0.018). CONCLUSION: We did not find a significant association between the MnSOD, GSTM1 and GSTP1 polymorphisms and carotid atherosclerosis. The GSTT1-0 genotype and GSTT1-0/GSTM1-0 haplotype might be a potential determinants of susceptibility to advanced atherosclerosis in patients with type 2 diabetes mellitus.

Serum cystatin C levels in normal pregnancy.

AIM: The aim of this study was to determine the levels of cystatin C, creatinine and creatinine clearance in different trimesters of uncomplicated pregnancy in women with normal kidney function. SUBJECTS AND METHODS: A total of 109 pregnant women were included: group 1 - 38 women (average age 29.63 ± 4.3 y) in the first trimester, Group 2 - 32 women (average age 33.56 ± 5.95 y) in the second trimester and Group 3 - 39 pregnant women (average age 30.1 ± 6.95 y) in the third trimester. Serum cystatin C was determined by the PENIA method (Particle-Enhanced Nephelometric Immuno-Assay), using Behring tests (Behring Diagnostics GmbH, Marburg, Germany). Results were statistically analyzed using the ANOVA. RESULTS: A statistically significant increase in serum cystatin C level was found in the third trimester of pregnancy (0.69 ± 0.16 mg/l vs. 0.78 ± 0.26 mg/l vs. 1.21 ± 0.30 mg/l). CONCLUSION: It appears that cystatin C is not a reliable marker of kidney function in pregnancy and that its increase is connected with a combination of several factors, including endotheliasis, hormonal influence and glomerular filtration rate (GFR) alterations.

Interleukin-18 Promoter Gene Polymorphisms are not Associated with Myocardial Infarction in Type 2 Diabetes in Slovenia.

Type 2 diabetes is a major risk factor for myocardial infarction (MI) and chronic inflammation may play a central role in both diseases. Interleukin (IL)-18 is a potent proinflammatory cytokine, which is considered important in acute coronary syndromes and type 2 diabetes. We investigated the association of the -137 (G>C), polymorphism (rs187238) and the -607 (C>A) polymorphism (rs1946518) of the IL-18 gene promoter region in 495 Caucasians with type 2 diabetes, of whom 169 had MI and 326 subjects had no clinically evident coronary artery disease (controls). We also investigated the impact of these polymorphisms on the serum IL-18 level in subsets of both groups and in a normal group. Genotype distributions of the polymorphisms showed no significant difference between cases and controls. However, IL-18 serum levels were significantly lower in diabetics with the 137 CC genotype than in those with other genotypes (241.5 ± 132.7 ng/L vs. 340.2 ± 167.4 ng/L; p <0.05). High sensitivity C-reactive protein and IL-18 serum levels were higher in diabetics in the MI group than in the control group. We conclude that these IL-18 promoter gene polymorphisms are not risk factors for MI in Caucasians with type 2 diabetes.

Oral status in patients receiving 5-fluorouracil for colorectal cancer.

PURPOSE: Oral complications are frequent and troublesome symptoms for those undergoing chemotherapy for cancer. Several antineoplastic agents are proved to have stomatotoxic potential, among them 5-fluorouracil (5-FU). The aim of the present study was to evaluate the oral status and patient experiences during chemotherapy with 5-FU for colorectal cancer. METHODS: Twenty-eight patients treated with 5-day 5-FU plus leucovorin entered this study. Positive data about oral symptoms were taken by anamnesis. Mucositis severity index, gingival index, plaque index, probing pocket depth and bleeding on probing have been used to assess oral mucosa and periodontal status of the patients. Patients were examined prior to chemotherapy and 14 days after the start of the chemotherapy cycle. RESULTS: Mild to moderate subjective complaints concerning oral cavity were reported by 17.9% of patients before and 39.2% of patients after chemotherapy. Clinical examination revealed oral mucosa damage in 10.7% and 35.7% of patients, with mean mucositis score of 0.14 and 0.54 before and after chemotherapy, respectively. Although mean values of all periodontal indices were elevated after chemotherapy, only increase in gingival index was statistically significant (p=0.035). Mucositis was significantly correlated with oral pain (p=0.00), xerostomia (p=0.00), and plaque index (p=0.077), while the correlation between mucositis and the rest of the examined parameters was not significant. CONCLUSION: Oral complications were not highly expressed in this study. Although 5-FU is considered to exert significant stomatotoxic effect, severe mucositis was far less common in this study compared to studies reported elsewhere.

Cardiotoxic injury caused by chronic administration of microcystin-YR.

Microcystins are cyclic peptide toxins. Chronic intoxication with well-known members of the microcystin family--microcystins-LR--induces liver tumour formation, injury of kidney and heart. Despite worldwide distribution in the environment, the effects of microcystins-YR have not been studied extensively. The aim of the study was to evaluate whether microcystins-YR, in relatively low doses, have a toxic effect on cardiomyocytes of chronically treated rats. Male adult Wistar rats were treated every second day for 8 months with microcystins-YR (10 microg/kg i.p., N = 5). Control groups were treated either with vehicle (ethanol and methanol 4 : 1 v/v; N = 5) or with physiologic saline (N = 4). The heart sections of microcystin-YR-treated rats revealed decreased volume density of cardiac muscle tissue (microcystins- YR = 0.485 mm3/mm3 +/- 0.003; vehicle = 0.493 mm3/mm3 +/- 0.002; saline = 0.492 mm3/mm3 +/- 0.002) due to fibrous proliferation. A few lymphocyte infiltrates were observed. Most of cardiomyocytes were enlarged (microcystins-YR = 20.19 microm +/- 1.34, vehicle = 17.45 microm +/- 0.52, saline = 16.00 microm +/- 1.43), with enlarged and often bizarre-shaped nuclei and decreased myofibril volume fraction (microcystins- YR = 0.416 mm3/mm3 +/- 0.009; vehicle = 0.472 mm3/ mm3 +/- 0.009; saline = 0.479 mm3/mm3 +/- 0.010). No TUNEL-positive cells were found in the heart sections of rats in all groups. The results allow the conclusion that chronic exposure to low doses of microcystins-YR may cause atrophy and fibrosis of the heart muscle.