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Introduction: The prognosis of rectal cancer (RC) is influenced by various clinical, pathological, and molecular factors. Recent research has focused on developing predictive models to improve the accuracy of survival predictions. The CDH1 gene, encoding the Ecadherin protein, plays a significant role in cell adhesion and has been implicated in various cancers, including colorectal cancer (CRC). Current research highlights its potential as a predictive marker for CRC diagnosis and treatment, although its role is complex and multifaceted, specifically in RC as a distinct entity. CDH1 gene expression is generally reduced in CRC, correlating with decreased E-cadherin levels. This reduction is more pronounced as the tumor progresses, suggesting its potential as a biomarker for disease progression. Aim: This study aimed to assess the prognostic role of CDH1 gene expression in a selected cohort of RC patients. Methods: We conducted a preliminary study on 37 untreated RC patients who underwent curative resection from 2016 to 2018 at the Oncology Institute of Vojvodina. Relative expression of CDH1 mRNA was measured using quantitative real-time PCR. A receiver operating characteristic (ROC) curve was used to measure the possibility of using CDH1 expression as a biomarker of metastasis in RC. Results: For CDH1 expression, ROC analysis showed good diagnostic accuracy in metastases prediction (AUC = 0.700; SE = 0.088; 95% CI = 0.527–0.839; P = 0.023). The cutoff value was ≤0.105330854, sensitivity was 100%, and specificity was 40%. Moreover, ROC analysis also demonstrated good prognostic accuracy of CDH1 expression in terms of death outcome prediction (AUC = 0.686; SE = 0.089; 95% CI = 0.513–0.828; P = 0.037). The cutoff value was ≤0.105330854, sensitivity was 100%, and specificity was 43.48%. Conclusion: Obtained results demonstrated a strong prognostic role of CDH1 expression in RC metastases and outcome prediction., Uvod: Na prognozu karcinoma rektuma (KR) utiču različiti klinički, patološki i molekularni faktori. Nedavna istraživanja su se fokusirala na razvoj prediktivnih modela kako bi se poboljšala tačnost predviđanja ishoda bolesti. Gen CDH1, koji kodira protein E-kadherin, igra značajnu ulogu u ćelijskoj adheziji i učestvuje u progresiji različitih tipova tumora, uključujući kolorektalni karcinom (KRK). Sadašnja istraživanja naglašavaju njegov prediktivni potencijal u dijagnostici i lečenju KRK, iako je njegova uloga složena i višestruka, posebno u KR kao posebnom entitetu. Ekspresija CDH1 gena je generalno smanjena u KRK, što je u korelaciji sa smanjenim nivoima proteina E-kadherin. Ovo smanjenje je izraženije kako tumor napreduje, što ukazuje na njegov potencijalnu ulogu kao biomarkera progresije bolesti. Cilj: Cilj ove studije je procena prognostičke uloge ekspresije CDH1 gena u odabranoj kohorti pacijenata sa KR. Metode: Ova preliminarna studija obuhvatila je 37 netretiranih KR pacijenata koji su lečeni operativnim putem u periodu 2016-2018. god. u Institutu za onkologiju Vojvodina. Ekspresija CDH1 iRNA izmerena je kvantitativnom PCR metodom u realnom vremenu. ROC (receiver operating characteristic) analiza krive korišćena je za procenu prognostičke uloge CDH1 vrednosti ekspresije kao biomarkera metastaza u RC. Rezultati: U slučaju ekspresije CDH1 gena, ROC analiza pokazala je dobar prognostički skor u predikciji pojave metastaza (AUC = 0.700; SE = 0.088; 95% CI = 0.527–0.839; P = 0.023). Izmerena “cutoff” vrednost bila je ≤0.105330854, senzitivnost 100% i specifičnost 40%. Pored toga, ROC analiza je dodatno pokazala dobru prognostičku preciznost ekspresije CDH1 gena u predikciji krajnjeg ishoda bolesti (AUC = 0.686; SE = 0.089; 95% CI = 0.513–0.828; P = 0.037). Dobijena je “cutoff” vrednost ≤0.105330854, sa senzitivnošću od 100%, dok je specifičnost bila 43.48%. Zaključak: Dobijeni rezultati su pokazali snažnu prognostičku ulogu ekspresije CDH1 u predikciji pojave me

Myocardial infarction (MI) is followed by left ventricular (LV) remodeling, the process crutial in preserving heart structure and function. High mobility group box 1 (HMGB1) gene has been investigated in inflammatory reparative processes, whereas serum HMGB1 has been recognized as a candidate biomarker for acute MI. The aim of this preliminary study was to investigate the potential effect of HMGB1 mRNA expression on post-MI heart remodeling. [etc]

Recently, we investigated targeted expression of ferroptosis-related genes in relation to multiple sclerosis (MS) severity, as it is proposed to be one of the mechanisms that cause neurodegeneration, hallmark of progressive multiple sclerosis (MS). The upregulation of cyclin dependent kinase inhibitor 1A (CDKN1A), a gene responsible for reduction in sensitivity to ferroptosis, was found in the secondary progressive (SP) compared to relapse remitting (RR) MS patients. We aim to investigate genetic variant rs3176326 (G/A), which has been marked by an ENCODE as the most probable functional variant in this gene, in association with SP MS. The study included a total of 213 MS patients (159 RR and 54 SP) from Serbia. Diagnosis of clinically definite MS was performed according to the revised McDonald criteria. Genotyping has been performed by TaqMan® technology. We found significantly higher frequency of rare allele bearing genotypes according to dominant model (AG+AA vs. GG) in SP MS compared to RR MS patients (p=0.015). Genotypes bearing rare allele were significantly and independently associated with SP MS with an adjusted OR 2.08 (95% CI 1.10 - 3.95, p=0.02). OR was adjusted for sex and active smoking. There was no significant association with EDSS or MSSS scores (p>0.05). Our preliminary results suggest possible association of CDKN1A rs3176326 with MS severity. Further replication of the results on larger sample size and functional analysis are inevitable.

The prognosis of rectal cancer (RC) can be influenced by various factors, including the location of the tumor. Studies have shown that the position of the tumor within the rectum can impact the recurrence and overall survival rates. Tumor localization in RC is also associated with gene expression alterations. ZEB1 is a transcriptional factor that promotes tumor invasiveness via regulation of the epithelialmesenchymal transition process and plays a significant role in RC by influencing tumor aggressiveness. This study aimed to evaluate the location-specific expression of the ZEB1 gene and its prognostic role in RC patients. This preliminary study included 45 untreated RC patients who underwent curative resection. In 10 patients, the tumor was located in the low rectum (0-5 cm), while 35 tumors were positioned within the mid and upper rectum (5-15 cm). We isolated mRNA from postoperative tumor FFPE samples. TaqMan® gene expression assay was used for quantitative real-time PCR analysis of ZEB1 gene expression. The preliminary analysis revealed that relative ZEB1 gene expression was significantly higher in upper RC locations than among low-positioned tumors (0.00198 vs. 0.000947, p=0.024). We did not find a significant difference in overall survival (OS) and disease-free survival (DFS) among RC patients based on the tumor location and ZEB1 expression (p > 0.05 in both cases). However, among 35 patients with upper-positioned tumors, we noted that patients with elevated ZEB1 expression had shorter DFS than patients with low ZEB1 expression (48.062±6.557 vs. 61.682±6.095 months), although the observed difference was not statistically significant (p=0.270). Our results indicate the importance of evaluating spatial ZEB1 expression in RC and suggest its potential prognostic role, especially in determining the survival time without disease recurrence in RC patients. However, further research on a larger number of samples is needed to confirm these findings.

Background/Aim. The renin-angiotensin system (RAS) is linked to the development of atherosclerosis (As), including its initiation and progression. Besides the well-known angio-tensin-converting enzyme (ACE), two newer RAS family members are related to vascular remodeling - ACE2 as a homolog of ACE and collectrin [transmembrane protein 27 (TMEM27)] as a homolog of ACE2. Up to now, a limited number of studies have examined the expression of these RAS components in advanced carotid plaque (CP) tissue based on the sex of the patients and plaque phenotypes (PPs). There are two ultrasonographically defined PPs - the hypoechogenic plaque (HoP) and the hyperechogenic plaque (HerP) phenotype. The aim of the study was to investigate whether there was a correlation between the expression of RAS components in the CP and the sex and PPs of patients. Methods. We examined 74 patients with advanced CP who underwent carotid endarterectomy. The intraplaque expression of RAS components was determined with the real-time polymerase chain reaction, using the TaqMan® gene expression assays and Western blot. A two-way ANOVA followed by a post-hoc Tukey test was performed for the statistical analysis of results. Results. No interaction was recorded between the sex of the patients and PPs in influencing the relative expression of ACE and TMEM27 messenger RNA (mRNA) (p > 0.05). In 56.06% of plaque samples, no expression of ACE2 mRNA was detected. Among the plaques where ACE2 mRNA expression was detected, its expression level was higher in females with the HoP phenotype compared to females with the HerP phenotype (p < 0.001). In patients with the HoP phenotype, females had higher expression of ACE2 mRNA than males (p < 0.05). In the male study group, ACE protein levels were significantly lower in the HoP phenotype compared to the HerP phenotype (p < 0.001). Fe-males with the HoP phenotype had significantly higher ACE protein levels than males with the HoP phenotype (p < 0.0001). Conclusion. Ou, Renin-angiotenzin sistem (RAS) povezan je sa razvojem ateroskleroze (As), uključujući njen nastanak i progresiju. Pored dobro poznatog angiotenzinkonvertujućeg enzima (angiotensin-converting enzyme – ACE), dva nova člana RAS familije povezana su sa remodelovanjem zidova krvnih sudova – ACE2 kao homolog ACE i kolektrin [transmembrane protein 27 (TMEM27)] kao homolog ACE2. Do sada je mali broj studija ispitivao ekspresiju komponenti RAS sistema u tkivu uznapredovalog karotidnog plaka (KP) u odnosu na pol bolesnika i fenotip plaka (FP). Postoje dva tipa KP definisana primenom ultrazvuka – fenotip hipoehogenog plaka (HoP) i fenotip hiperehogenog plaka (HerP). Cilj rada bio je da se ispita da li postoji korelacija između ekspresije komponenti RAS u KP i pola i FP bolesnika. Metode. Ispitano je 74 bolesnika sa uznapredovalim KP koji su bili podvrgnuti operativnoj proceduri karotidne endarterektomije. Ekspresija komponenti RAS sistema u tkivu plaka utvrđena je lančanom reakcijom polimeraze u realnom vremenu (real-time polymerase chain reaction)primenom eseja TaqMan® tehnologije i metode Western blota. Dvosmerna analiza varijanse i Tukey post-hoc test korišćen su za statističku obradu rezultata. Rezultati. Nije utvrđena interakcija FP i pola bolesnika u uticaju na relativnu ekspresiju informacione RNK (iRNK) za ACE i TMEM27 (p > 0,05). U 56,06% uzoraka plaka nije detektovana ekspresija iRNK za ACE2. U plakovima u kojima je detektovana ekspresija iRNK za ACE2, njen nivo bio je viši kod žena sa HoP u poređenju sa ženama sa HerP (p < 0,001). U grupi bolesnika sa fenotipom HoP, žene su imale viši nivo iRNK za ACE2 nego muškarci (p < 0,05). U grupi muškaraca, nivoi ekspresije ACE proteina bili suznačajno niži u fenotipu HoP u poređenju sa HerP (p < 0,001). Žene sa fenotipom HoP imale su značajno viši nivo ACE proteina u poređenju sa muškarcima sa HoP (p < 0,0001). Zaključak. Naši rezultati pokazali su da postoje promene nivoa ekspresije ACE i ACE2, na nivou proteina i iRNK u uzn

Background The MMP-9 is a known player in atherosclerosis, yet associations of the MMP-9 -1562 C/T variant (rs3918242) with various atherosclerotic phenotypes and tissue mRNA expression are still contradictory. This study aimed to investigate the MMP-9 -1562 C/T variant, its mRNA and protein expression in carotid plaque (CP) tissue, as a risk factor for CP presence and as a marker of different plaque phenotypes (hyperechoic and hypoechoic) in patients undergoing carotid endarterectomy. The MnSOD as an MMP-9 negative regulator was also studied in relation to CP phenotypes. Methods and results Genotyping of 770 participants (285 controls/485 patients) was done by tetra-primer ARMS PCR. The MMP-9 mRNA expression in 88 human CP tissues was detected by TaqMan® technology. The protein levels of MMP-9 and MnSOD were assessed by Western blot analysis. The MMP-9 -1562 C/T variant was not recognized as a risk factor for plaque presence or in predisposing MMP-9 mRNA and protein levels in plaque tissue. Patients with hypoechoic plaques had significantly lower MMP-9 mRNA and protein levels than those with hyperechoic plaque (p = 0.008, p = 0.003, respectively). MnSOD protein level was significantly higher in hypoechoic plaque compared to hyperechoic (p = 0.039). MMP-9 protein expression in CP tissue was significantly affected by sex and plaque type interaction (p = 0.009). Conclusions Considering the differences of MMP-9 mRNA and protein expression in CP tissue regarding different plaque phenotypes and the observed sex-specific effect, the role of MMP-9 in human atherosclerotic plaques should be further elucidated.

Introduction: Venous invasion has consistently been shown to be associated with poor prognosis in rectal carcinoma (RC), both when detected by pathology and radiology. Extramural venous invasion (EMVI) is characterized by the presence of tumor cells within veins outside the bowel wall and is strongly associated with poor survival and increased risk of local recurrence and distant metastases. Molecular basis of EMVI is still unexplored and genes that regulate tumor microenvironment interactions may have significant role in this process. ZEB1 is transcriptional factor that promote cancerogenesis by indirect regulation of epithelial-mesenchymal transition (EMT) process, while LOXL2 contributes to tumor invasion and metastasis due to its role in the stabilization of the extracellular matrix. Aim: This study aimed to compare the expression level of ZEB1 and LOXL2 genes in relation to EMVI status and other clinic-pathological parameters of RC patients. Methods: We conducted preliminary study on 21 untreated RC patients (9 EMVI+ and 11 EMVI- ) who underwent curative resection in 2016-2018 at Oncology Institute of Vojvodina. The presence of EMVI was assessed on standard hematoxylin and eosin-stained histolological sections of postoperative tumor specimen samples, from which RNA was isolated. Expression of ZEB1 and LOXL2 mRNA was measured using quantitative real-time PCR. Results: Comparative analysis revealed higher expression level of ZEB1 in EMVI positive samples and in patients in TNMIII stage, however the observed differences had no statistic significance (p=0.323 and p=0.197, respectively). Significant difference in LOXL2 expression according to the EMVI status was not detected (p=0.915), while we noted higher LOXL2 expression in late stages of disease, but without statistic significance (p=0.342). Relative expression of these two genes was not associated with metastases frequency and death outcome. Conclusion: Further analyses on larger number of samples with more pot, Uvod: Venska invazija je kontinuirano asocirana sa lošom prognozom kod obolelih od karcinoma rektuma (KR), bilo da je detektovana patološkim ili radiološkim metodama. Ekstramuralna venska invazija (EMVI) se karakteriše kao prisustvo tumorskih ćelija u venskim sudovima izvan zida debelog creva koje je značajno asocirano sa lošim preživljavanjem i povećanim rizikom za nastanak lokalnih recidiva i udaljenih metastaza. Molekularna osnova EMVI procesa nije dovoljno ispitana, a geni koji regulišu interakcije u tumorskoj mikrosredini mogu imati potencijalnu ulogu. ZEB1 je transkripcioni factor koji stimuliše kancerogenezu indirektnom regulacijom epitelnomezenhimske tranzicije (EMT), dok LOXL2 doprinosi procesu tumorske invazije ulogom u stabilizaciji ektracelularnog matriksa. Cilj: Cilj ove studije je uporediti relativnu ekspresije gena ZEB1 i LOXL2 u odnosu na EMVI status i druge kliničko-patološke parametre KR pacijenta. Metode: Ova preliminarna studija obuhvatila je 21 netretiranih KR pacijenata (9 EMVI+ i 11 EMVI-) koji su lečeni operativnim putem u periodu 2016-2018. god. u Institutu za onkologiju Vojvodina. Prisustvo EMVI je utvrđeno na standardno hematoksilinom i eozinom bojenim isečcima postoperativnog tumorskog tkiva iz kojih je izolovana RNK. Ekspresija ZEB1 i LOXL2 iRNA izmerena je kvantitativnom PCR metodom u realnom vremenu. Rezultati: Uporednom analizom uočena je povišena ekspresija ZEB1 kod EMVI+ uzoraka i kod obolelih u TNMIII stadijumu, ali uočene razlike nisu bile statistički značajne (p=0,323 i p=0,197, respektivno). Znčajna razlika u ekspresiji LOXL2 u odnosu na EMVI status nije detektovana (p=0,915), a zabeležena je i povećana ekspresija LOXL2 u kasnim stadijumima bolesti, ali bez statističke značajnosti (p=0,342). Relativna ekspresija ova dva gena nije značajno povezana sa pojavom metstaza i krajnjim ishodom bolesti. Zaključak: Dalje analize na većem broju uzoraka sa više uključenih molekularnih targeta u studiju su neophodne i planirane u budućnosti.

Glutathione S-transferases (GSTs) are the family of enzymes involved in the second line of defense against oxidative stress (OS). The lack of GSTT1/GSTM1 enzyme quantity or activity, due to the presence of homozygous deletion compromises antioxidative defense resulting in OS. OS is the critical mechanism in the pathophysiology of atherosclerosis, coronary artery disease, and myocardial infarction (MI). The increase in reactive oxygen species together with the process of apoptosis plays a role in left ventricular remodeling (LVR) after MI. The associations of GSTT1 and GSTM1 gene polymorphisms with the risk of MI are inconsistent. The aim was to analyze the association of GSTT1/GSTM1 null genotypes with first MI and LVR 8 months after the MI. The study involved 330 controls and 438 consecutive patients with symptoms and signs of first MI. The subgroup of 150 MI patients was prospectively followed up for 6 months. Evidence of maladaptive LVR was obtained by 2D Doppler echocardiography 3-5 days and 6 months after the MI. A multiplex polymerase chain reaction was used to detect the deletion in GSTT1 and GSTM1 genes. GSTM1 null genotype was significantly and independently associated with first MI (adjusted OR = 1.45 95% CI 1.03-2.03, p = 0.03). Association of double null genotypes with maladaptive LVR in patients 6 months after the first MI was no longer significant after adjustment for factors that differed significantly between patients with and without maladaptive LVR. This study demonstrated the association of GSTM1 null genotypes with the risk of MI in the Serbian population.

Foods and food products that contain polyphenols are proposed to modulate risk of cardiovascular disease. The aim of this three-arm, crossover, randomized, double-blind, placebo-controlled intervention study was to examine the impact ofAronia melanocarpajuice (AMJ), high-polyphenol (AMJ treatment, 1.17 g/100 mL polyphenols) and low-polyphenol (dAMJ treatment, 0.29 g/100 mL polyphenols) dose, on the transcriptome in peripheral blood mononuclear cells (PBMC) of 19 subjects at cardiovascular risk. Transcriptome data were obtained by microarray. Bioinformatic functional annotation analysis was performed on both the whole transcriptome datasets and the differentially expressed genes (DEGs). Expression of selected DEGs was validated by RT-qPCR. Administration of AMJ and dAMJ treatments during the two consecutive four-week treatment periods had additive effects on PBMC transcriptome profiles, with the most pronounced and specific effect noticed for AMJ in the last treatment period (TP3) of the trial. Between the high-dose and low-dose treatments in TP3, there was a multitude of overlapping DEGs and DEG-enriched biological processes and pathways, which primarily included immunomodulation and regulation of cell proliferation/death. Increased expression ofTNF,IL1B,IL8,RGS1,OSM, andDUSP2in TP3 was confirmed by RT-qPCR. The results suggest the immunomodulatory effects of prolonged habitual consumption of polyphenol-rich aronia juice in individuals at cardiovascular risk.

Foods and food products that contain polyphenols are proposed to modulate risk of cardiovascular disease. The aim of this three-arm, crossover, randomized, double-blind, placebo-controlled intervention study was to examine the impact of Aronia melanocarpa juice (AMJ), high-polyphenol (AMJ treatment, 1.17 g/100 mL polyphenols) and low-polyphenol (dAMJ treatment, 0.29 g/100 mL polyphenols) dose, on the transcriptome in peripheral blood mononuclear cells (PBMC) of 19 subjects at cardiovascular risk. Transcriptome data were obtained by microarray. Bioinformatic functional annotation analysis was performed on both the whole transcriptome datasets and the differentially expressed genes (DEGs). Expression of selected DEGs was validated by RT-qPCR. Administration of AMJ and dAMJ treatments during the two consecutive four-week treatment periods had additive effects on PBMC transcriptome profiles, with the most pronounced and specific effect noticed for AMJ in the last treatment period (TP3) of the trial. Between the high-dose and low-dose treatments in TP3, there was a multitude of overlapping DEGs and DEG-enriched biological processes and pathways, which primarily included immunomodulation and regulation of cell proliferation/death. Increased expression of TNF, IL1B, IL8, RGS1, OSM, and DUSP2 in TP3 was confirmed by RT-qPCR. The results suggest the immunomodulatory effects of prolonged habitual consumption of polyphenol-rich aronia juice in individuals at cardiovascular risk.

BACKGROUND: Breast carcinoma is heterogeneous disease. Understanding the process of invasion and metastasis and the selection of the therapy for patients with breast carcinomas still remains difficult. MicroRNAs are powerful gene expression regulators. Because of inconsistent findings, we have analyzed potential difference in miR-155 levels in three breast cancer groups. OBJECTIVES: Our goals were to examine miR-155 expression levels in normal tissue, non-invasive and invasive breast carcinomas, and their association with standard clinical and pathological parameters and oncomiR-21, and to investigate the ability of miR-155 to separate invasive breast carcinomas with non-invasive component from pure invasive. METHODS: In the group of 40 breast tissue samples, relative expression levels of miR-155 were examined with stem-loop quantitative real-time PCR using TaqMan technology. RESULTS: The significant difference among four examined groups of the breast tissue was detected (p = 0.001). In the group of pure invasive tumors, patients with positive nodal status had significantly higher miR-155 levels (p = 0.046). CONCLUSION: Our results suggest that miR-155 might be involved in breast cancer pathogenesis and in tumor spreading to the lymph nodes, and that it might be used as biomarker for additional stratification of patients with invasive breast carcinomas with non-invasive component.

Background: The angiotensin II type 2 receptor (AT2R) - 1332 A/G polymorphism has been denoted as functional and associated with certain cardiovascular disease phenotypes. However, there are no studies considering the association of this gene polymorphism with carotid atherosclerosis (CA) and cerebrovascular events. Therefore, the aim of our study was to investigate a possible association of the AT2R - 1332 A/G polymorphism with the occurrence of carotid plaques (CPs) and history of cerebrovascular insult (CVI) in advanced CA. Methods: The study group included 381 controls and 509 patients with CA consecutively admitted for endarterectomy. Genotyping was determined by polymerase chain reaction-restriction fragment length polymorphism method. The association was analyzed separately for males and females because the AT2R gene is located on the X chromosome. Results: The AT2R - 1332 GG genotype was associated with the advanced CA in the female study group (recessive model of inheritance, AA+AG versus GG; adjusted odds ratio [OR] = 2.25; 95% confidence interval [CI] 1.17-4.33; P=.01). In the male subgroup of patients with CA, the significant overrepresentation of G/- hemizygote was detected in patients with CVI compared to male patients without this event (crude OR = 2.05, 95% CI 1.20-3.50, P=.008). Conclusions: This study suggests a gender-specific association between the AT2R -1332 A/G polymorphism and the occurrence of CP and the history of CVI in advanced CA, but further replication studies are needed. (C) 2016 National Stroke Association. Published by Elsevier Inc. All rights reserved.

Objectives: The objectives were to detect and compare the expression of toll-like receptors (TLRs) 2, 4 and nuclear factor kappa B in mucosal lesions of chronic otitis. Methods: Fifty-five tissue samples obtained from children and adults operated on for otitis were investigated by semiquantitative immunohistochemical methods using polyclonal antibodies for TLR 2, 4 and NFkB. Kruskal-Wallis, Mann-Whitney, and Kendalls tau rank correlation tests were used. Results: Stronger expression of TLR2, 4 was found in inflamed mucosa than in the control for children and adults (TLR2: H = 23.86, P LT .001; TLR4: H = 22.80, P LT .001) (TLR2: H = 17.53, P LT .001; TLR4: H = 11.99, P LT .001); in cholesteatoma perimatrix compared to tubotympanic lesions in children (TLR2: H = 11.06, P = .004; TLR4: H = 10.61, P = .005) and adults (TLR2: H = 10.73, P =.013; TLR4: H = 9.65, P = .021). No differences were found in NFkB expression (H = 0.042, P = .99). Significant correlations were found for all pairs of molecules in cholesteatoma and tubotympanic mucosa of adults (TLR2, 4: P = .002, P LT .001; TLR2-NfkB: P = .032, P = .021; TLR4-NFkB: P =.035, P = .0013), only TLR4-NFkB in tubotympanic otitis of children (P = .026). Conclusions: Toll-like receptors 2, 4 and NFkB mediate inflammation in cholesteatoma and mucosal lesions of tubotympanic otitis in children and adults. Significant correlations between all pairs of molecules in all samples were detected in adults, but only TLR4-NFkB in children.

Toll like receptors (TLRs) are the pattern recognition receptors, important for the innate immune system, the first line of defense against bacterial infections which are the main cause of otitis media (OM). Our aim was to investigate the association of TLR4 (Thr399Ile and Asp299Gly) and TLR2 (Arg753Gln) most common polymorphisms with OM prevalence as well as with morphologic changes of middle ear during chronic otitis media. We had 111 controls and 186 patients divided into three groups of COM: nonsuppurative OM, suppurative OM and cholesteatoma. Frequencies of the genotypes of the TLR 2 gene polymorphism were significantly different between men and women having nonsuppurative OM (p=0.03).

Karotidna ateroskleroza (KA) je hronična bolest koja započinje aktivacijom i nakupljanjem inflamatornih ćelija koje se pune lipidima u zidu arterijskog krvnog suda. Aterosklerotski plak sadrži lipidno jezgro, fibroznu kapu, akumulira glatke mišićne ćelije (GMĆ) i proteine ekstraćelijskog matriksa (EĆM).Renin-angiotensin system (RAS) ima ključnu ulogu u aktivaciji inflamacije u zidu krvnog suda. Glavni molekuli RAS su: angiotenzin-konvertujući enzim (ACE), angiotenzin-konvertujući enzim-homolog (ACE2), angiotenzin II receptori tipa 1 (ATR1) i tipa 2 (ATR2), ACE2 homolog (TMEM 27) i regulatorna RNK (miR-155). Angiotenzin II (Ang II) je glavni efektorni molekul RAS koji se sintetiše aktivnošću ACE, a razgrađuje aktivnošću ACE2. Ang II ostvaruje svoje efekte vezujuću se za ATR1 i ATR2. Efekti Ang II imaju važnu ulogu u regulaciji vaskularne homeostaze i to su: vazokonstrikcija, migracija, proliferacija i hipertrofija GMĆ, povećana sinteza EĆM i povećana produkcija matriks-metaloproteinaza (MMPs). Ekspresija ATR1 regulisana je sa miR-155. Balans između aktivacije i represije RAS može biti odlučujući u patološkom remodelovanju zida krvnog suda i patogenezi KA. Zato je važno ispitati funkcionalnu aktivaciju RAS sistema na lokalnom (tkivnom) nivou u karotidama i to na nivou ekspresije gena koji kodiraju za komponente RAS, enzime i receptore. Ekspresija gena je regulisana preko prisustva/odsustva različitih alelnih varijanti funkcionalnih polimorfizama u genima za RAS.Genetičko-epidemiološka studija gena RAS je urađena po pacijenti - kontrole dizajnu, na tri gena i tri polimorfizma. Ova studija obuhvatila je 750 ispitanika oba pola iz populacije Srbije, 505 pacijenata i 246 kontrola iste etničke pripadnosti. Urađena je studija asocijacije polimorfizma I/D u genu za ACE, polimorfizma A1166C u genu za ATR1 i polimorfizma A/G -1332 u genu za ATR2 sa nastankom KA, kliničkim parametrima KA, fenotipom plaka (stabilni (SAP) i nestabilni (NAP)), kliničkim događajima (prolazni ishemi, The carotid atherosclerosis (CA) is chronic disease, which begins with activation inflammatory cells and accumulation lipid in these cells within blood vessel wall. Atherosclerotic plaque containing a lipid core, fibrous cap, accumulates smooth muscle cells (GMC) and proteins of the extracellular matrix (ECM).The renin-angiotensin system (RAS) has a key role in activation of inflammation within blood vessel wall. The main components of the system are: angiotensin I-converting enzyme (ACE), angiotensin converting enzyme homolog (ACE2), angiotensin II receptor type 1 (ATR1), angiotensin II receptor type 2 (ATR2), ACE2 homolog (TMEM27) and regulatory RNA (miR-155). Angiotensin II (Ang II) is the main effector molecule of the RAS, which is synthesized by the activity of ACE and is cleaved by ACE2. Ang II achieves its effects by binding to ATR1 and ATR2. These effects has an important role in the regulation of vascular homeostasis and include the following: vasoconstriction, vascular smooth muscle cells (SMC) migration, proliferation and hypertrophy, increased synthesis of extracellular matrix (ECM) and enhanced production of matrix metalloproteinases (MMPs). The expression of ATR1 is regulated by the miR-155. The balance between activation and repression of RAS can be decisive in pathological remodeling of the vessel wall and the pathogenesis CA. It is therefore important to examine the functional activation of RAS at the local (tissue) level in carotid vessel wall at level of expression genes encoding for components of the RAS, enzymes and receptors. The gene expression of components of RAS is regulated through the presence/absence of different allelic variants of functional polymorphisms in these genes.The genetic epidemiological study for RAS genes was performed by case-control design and consisted of three genes and three polymorphisms within these genes. This study included 750 subjects of both sexes from the Serbian population, 505 patients and 246 controls of the

Introduction: The ACE I/D polymorphism was mostly investigated in association with intima-media thickness, rarely with severe atherosclerotic phenotype. Materials and methods: We investigated the association of I/D polymorphism with severe carotid atherosclerosis (CA) (stenosis GT 70%) in asymptomatic and symptomatic patients undergoing carotid endarterectomy. The 504 patients subjected to endarterectomy and 492 healthy controls from a population in Serbia were investigated as a case-control study. Results: The univariate logistic regression analysis revealed ACE DD as a significant risk factor for severe CA (odds ratio [OR] = 1.3, 95% confidence interval [CI] 1.0-1.7, p = 0.04). After adjustment for the common risk factors (age, hypertension, smoking, and HDL) ACE was no longer significant. However, we found a significant independent influence of DD genotype on plaque presence in a normotensive subgroup of patients (OR 1.8, CI 1.2-3.0, p = 0.01, corrected for multiple testing). In symptomatic patients D allele carriers were significantly more frequent compared with asymptomatic patients (OR 1.6 CI 1.0-2.6, p = 0.05). Conclusions: Our data suggests that ACE I/D is not an independent risk factor for severe CA. On the other hand, a significant independent genetic influence of ACE I/D appeared in normotensive and symptomatic patients with severe CA. This should be considered in further research toward resolving the complex genetic background of severe CA phenotype.

INTRODUCTION: Polymorphisms of renin-angiotensin system (RAS) in hypertensive patients may be associated with ECG parameters (QT, QRS complex, QRS axis, pNN50, rms SD). QT interval prolongation on 12-lead ECG is a predictor of sudden cardiac death due to ventricular arrhythmias (1). Hypertension is the state where RAS activity is modulated, and this can remodel the cardiac ion channels, resulting in prolongation (QT interval) and redirection of ventricular depolarization (QRS axis) and repolarization. Cardiac ion channels can be affected by such factors as the strech of the myocites by high blood pressure (2). To our knowledge no investigation has been done on the association of RAS genetic polymorphisms with the ECG parameters QT interval, QRS axis and time domain indexes of heart rate variability pNN50 and rmsSD. AIM. The aim of the study was to investigate the association of RAS genetic polymorphisms (ACE I/D, ATR1 1166 A/C, ATR2 1332 A/G) with ECG parameters (QT, QRS axis, pNN50 and rmsSD). METHODS. The study was conducted on 36 Serbian hypertensive untreated patients (age (mean±SD)=51.6±9.9 years, 17 female and 19 male) who were ECG monitored (Schiller multiparameter monitor) and taken whole blood sample for genetic analysis. DNA was extracted on the basis of phenol-chlorophorm method while genotypisation was done by PCR-restriction fragment lenght polymorphism method (3,4). Statistical analysis was performed using statistical package of SPSS program (SPSS 17.0). RESULTS. The results are summarized in Table 1. CONCLUSIONS: Our data are in line with the data in the literature that ACE D allele carriers intensify left ventricular hypertrophy, myocardial fibrosis and hypertension (5, 6). These changes contribute to electrophysiologic remodeling of myocardium resulting in change of parameters like QT interval and QRS axis. Parameters of short term heart rate variability showed tendency to be associated to RAS polymorphisms. Further investigations are needed to brin

Matrix metalloproteinases (MMPs) are involved in the remodeling of the extracellular matrix (ECM) in the arterial wall during atherogenesis. Collagens are the most abundant proteins in the ECM. MMP-8 is expressed by cells associated with the development of the atherosclerotic plaque. It cleaves collagen type I three times more potently than two other interstitial collagenases MMP-1 and MMP-13. The aim of this study was to investigate whether plasma MMP-8 values are associated with occurrence of carotid plaque (CP) and possible correlations with clinical and biochemical parameters in carotid atherosclerosis (CA) patients. Total plasma MMP-8 levels were quantified by ELISA in 63 patients with ultrasonographic evidence of CP presence and 12 controls. Plasma MMP-8 values were significantly higher in patients with CA compared with controls (median 23.36 ng/ml vs. 13.02 ng/ml, P LT 0.001) but they did not differ significantly according to gender, smoking and hypertensive status, associated diseases, and use of statins. Statistically significant positive correlations were observed between MMP-8 plasma values and C reactive protein (r=0.41, P=0.001), urea (r=0.50, P LT 0.001), aspartate transaminase (r=0.48, P=0.001), and creatinine levels (r=0.38, P=0.006). These results suggest association of MMP-8 plasma levels with occurrence of CP and correlation with certain biochemical markers. J. Clin. Lab. Anal. 24:246-251, 2010. (C) 2010 Wiley-Liss, Inc.

Multiple sclerosis (MS) is a complex inflammatory,demyelinating disease of central nervous system (CNS). All the essen-tial components of the renin-angiotensin system (RAS) are presentedin the mammalian brain. The angiotensin II (Ang II), biologicallyactive octapeptide is not only a vasoconstrictor, but also a pro-inflam-matory factor. Many of the classical and of the hypothetical functionsof brain Ang II are mediated by stimulation of AT1 receptors (AT1R).Brain AT2 receptors (AT2R) are highly expressed during development.In the adults, AT2R are restricted to areas predominantly involved inthe process of sensory information. The AT2R1332 A/G polymorph-ism was proposed to influence AT2R protein expression, and is themost studied polymorphism in this gene, in other diseases. Recently,the striking appearance of the RAS in MS brain was described.However, the role of AT2R remains to be clarified. Thus, the aim ofour study was to establish if there is an association between AT2R1332 A/G gene polymorphism and predisposition of MS Methods:Subjected group consisted of 122 female and 70 malepatients with MS and 75 female and 50 male controls from populationof Serbia. Genotyping was done by PCR and restriction digestion withEcoRI enzyme.Results:The genotype and allele frequencies for AT2R1332A/Ggene polymorphism are analyzed separately in females and males,since this gene is located on X chromosome. We detected significantoverrepresentation of1332A/G AA genotype (OR 1.6, 95% CI:1.0–2.7, p<0.05) in female patients with MS compared to female controls.In hemizygous males we didn’t found any difference between patientsand controls.Conclusion:The role of RAS genes in MS was neglected untilrecently. Than, it was shown that the role of RAS in the CNS isbeyond the regulation of cardiovascular function. Until now AT2R(1332A/G) gene polymorphism was widely studied and associatedwith hypertension and other vascular disease. Until now, there wereno studies concerning role of Ang II recept

The matrix metalloproteinase 9 (MMP-9) has been shown to play role in VSMC migration and proliferation by degradation of the basement membrane. MMP-9 expression is increased after vascular injury. The MMP-9 -1562 C/T gene polymorphism results in higher promoter activity of the T allele. The higher prevalence of –1562T allele was associated with coronary artery stenosis. The aim was to investigate the association of the MMP-9 -1562C/T polymorphism and susceptibility for development of carotid plaques. Genomic DNA was isolated from blood cells of 278 control subjects and 136 patients with carotid atherosclerosis. The genotypes were determined by tetra-primer ARMS PCR method previously developed by us and visualized by MADGE. We did not find the significant difference in genotype frequencies between controls: CC 70.82%, CT 27.05%, TT 2.14%, and patients: CC 74.26%, CT 25.74%, TT 0%. The allele frequencies didn’t differ as well (controls: C 0.84, T 0.16, patients: 0.87, 0.13. MMP-9 gene polymorphism -1562C/T is not the susceptibility factor for development of carotid atherosclerotic plaques in population of Serbia.

f chronic inflammation plays a causal role in atherogenesis, individuals with proinflammatory gene variants would be expected to develop atherosclerosis. The IL-6 gene were chosen because is an important component of the upstream inflammatory process and the -174G/C polymorphisms already implicated in cardiovascular disease. The aim of was to associate the IL-6 -174G/C polymorphism with development of carotid plaques. Materials and methods: Genomic DNA was isolated from blood cells of 278 controls and 136 patients with carotid atherosclerosis. The genotypes were determined by tetra-primer ARMS PCR and visualized by agarose gel electrophoresis. Results: The genotype distribution wasn’t significantly different in patients (GG 39.71%, GC 44.12%, CC 16.18%) compared to controls (GG 41.73%, GC 46.40%, CC 11.87%). Conclusion: The IL-6 gene polymorphism is not associated with advanced atherosclerotic phenotype such as plaque presence, in population of Serbia. As IL-6 is expressed in human atherosclerotic plaques there is the possibility that the level of its expression plays the more important role in this clinical phenotype than genotype of investigated polymorphism.

The effects of environmental exposures might be transmitted to cellular level through mechanisms that could vary between individuals. Gene polymorphism in GSTM1/GSTT1, both deletions in the gene, showed the significant influence in genetic association studies of air pollution. In order to analyze the association of GSTT1/GSTM1 polymorphism with respiratory and cardiovascular disease in regard to air- pollution, for the first time we detected the presence of GSTM1/GSTT1 deletion in two different environmental risk factors' populations, Belgrade and Obrenovac. The groups consisted of 71 (Belgrade)/89 (Obrenovac) participants. Genotyping was performed by multiplex PCR. The frequencies of “null” genotypes for GSTM1/GSTT1 were not significantly different between the two investigated groups (47.89%/44.94%, 15.49%/17.98%, respectively). In conclusion, the frequencies of detected “null” genotypes in investigated two population subsets of Serbian origin was similar and in range with the frequencies in other Caucasian populations.