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Arterial hypertension (AH) is a major independent risk factor for cardiovascular (CV) and cerebral mortality and morbidity and is one of the major risk factors for chronic kidney disease. Prehypertension (PHT) i.e. high normal arterial pressure (AP) is also associated with CV risk and risk of progression in AH. Previous studies of the genetic background of PHT have been insufficiently conducted, and the results indicate a possible association of certain genes involved in the etiopathogenesis of AH and PHT. The lack and sometimes inconsistency of information and results on the role of gene polymorphisms in the etiopathogenesis of PHT were the impetus to this research. Our hypothesis was that polymorphisms rs2004776 AGT, rs1799752 ACE, rs5196 AGTR1, rs1801253 ADRB1, rs13333226 UMOD and rs266729 and rs17300539 ADIPOQ, which are associated with AH and metabolic syndrome, are more common in individuals with high normal AP i.e. prehypertension than in individuals with optimal and normal AP. We chose these polymorphisms because these genes are involved in various mechanisms that lead to an increase in AP and metabolic complications. In this cross-sectional, observational study, 601 subjects were included ; 319 with high normal AP (PHT, 120/80–139/89 mmHg), both sexes, aged 20–45 years, and 282 subjects with normal and optimal AP as a control group (NT). The results showed that the investigated gene polymorphisms are not found more frequently in the PHT group than in the NT. However, a significantly lower frequency of the heterozygous genotype rs266729 ADIPOQ was found in PHT (35.1%), compared to the frequency in NT (44.7%), P = 0.03. The incidence of PHT is 0.66-fold lower in heterozygous (C/G) carriers of rs266729 ADIPOQ. We demonstrated a significant association of the minor allele G rs13333228 UMOD with lower values of systolic and diastolic AP and the minor allele A rs17300539 ADIPOQ with lower values of systolic AP. Additionally, our study resulted in the identification of two haplotypes that were significantly positively associated with PHT formation. The first is hIAGC with a 41% higher probability of developing PHT (rs1799752 ACE + rs13333226 UMOD + rs17300539 ADIPOQ + rs266729 ADIPOQ). The frequency of this haplotype is 8% higher in PHT than in NT. The second haplotype is hIAGCA and is 47% more likely to develop PHT (rs1799752 ACE + rs13333226 UMOD + rs17300539 ADIPOQ + rs266729 ADIPOQ + rs5186 AGTR1). The frequency of hIAGCA in NT is 18%, while in PHT it is 23%. Furthermore, the finding of a model of multivariate prediction of PHT (gene-gene-other determined parameters interaction) which includes a pro-prehypertensive significant effect of male sex, body weight, heart rate, fasting glucose concentration and total cholesterol is significant, while the protective effect within this model have body height and heterozygous (G/A) genotype rs17300539 ADIPOQ. We pointed to the possible importance of genes encoding proteins involved in essential biological pathways responsible for AP control, which include, among others, the renin-angiotensin-aldosterone system, the kidney, the central nervous system, blood vessels, and adipose tissue. In conclusion, the performed analyzes confirmed and showed the presence of significant gene - gene and gene - other determined parameters interactions (anthropometric, biochemical, environmental factors), but also gene - gene - other determined parameters in the etiology of PHT.

Hydrazones, represented by the general formula R1R 2C=N–NR3R 4 , are used in the synthesis of heterocyclic compounds, drug design, organocatalysis and coordination chemistry.[1, 2] These compounds exhibit interesting biological applications, such as anti- inflammatory, analgesic, antiplatelet, anticonvulsant, antitumoral, antiviral and antimicrobial activity.[1] The presented research is focused on the synthesis, characterization and biological activities of the selected aroylhydrazones. The condensation of salicylaldehyde derivative, namely 3- or 4- hydroxysalicylaldehyde, and hydrazide, specifically nicotinoyl- or isonicotinoylhydrazide, was achieved by conventional synthetic method (i.e. in the solution) as well as by the solventfree mechanochemical procedure. The synthesized compounds were characterized by the elemental and DSC analysis, IR and NMR spectroscopy. The molecular and crystal structures were determined by the single-crystal X-ray diffraction method. The in vitro cytotoxicity of hydrazones was investigated against THP-1 and HepG2 cells. Each cell line was treated with various concentrations of the tested compounds and the viability of cells after treatment was determined using MTS assay. Half maximal inhibitory concentration (IC50) values were calculated from dose‐response curves. Hydrazones were also tested for their in vitro antimicrobial activity against Gram-positive (Staphylococcus aureus and Enterococcus faecalis) and Gramnegative (Escherichia coli and Moraxella catarrhalis) bacteria by broth microdilution method. The results are reported in minimum inhibitory concentration (MIC) values.

Introduction: Low levels of melatonin (MLT) have been registered in patients with breast, prostate, lung, stomach and colon cancer. One of explanations for this was decreased sleep quality and increased fatigue in cancer patients. Although there is a significant improvement in oral cancer (OC) treatment in the last few years, a 5-year survival rate of these patients amounts to 60%. The aim of this research was to measure MLT in unstimulated whole saliva (UWS), stimulated whole saliva (SWS) and serum. Furthermore, the aim was to assess sleep quality using the Pittsburgh Sleep Quality Index (PSQI). Materials and methods: Altogether, 34 patients with OC and 33 sex and age matched healthy control subjects were included in this study. All samples were tested using ELISA commercially available kits. Results: Melatonin levels in UWS were significantly higher (P

Hydrazones, versatile ligands with the C=N–N group, and their metal complexes have been in the focus of interest due to applications in various fields, e.g. analytical chemistry, catalysis, pharmacy [1, 2]. In addition, these types of compounds have diverse biological activities such as cytotoxic activity [3]. However, literature reports on the in vitro cytotoxic activity of dioxomolybdenum(VI) complexes with hydrazones are very rare [4]. In this work, dioxomolybdenum(VI) complexes with 4‐methoxysalicylaldehyde isonicotinoyl hydrazone (H2L) were synthesized by the reaction of [MoO2(acac)2] and H2L with the addition of acid (HCl, HBr, HNO3 or H2SO4). The obtained [MoO2(HL)(MeOH)]X (X = Cl, Br, NO3, HSO4) compounds were characterized by elemental and thermogravimetric analysis, IR and NMR spectroscopy, and X‐ray diffraction methods (PXRD and SCXRD when suitable). The in vitro cytotoxicity of these compounds was investigated on human acute monocytic leukemia (THP‐1) and hepatocellular carcinoma (HepG2) cell lines. Each cell line was treated with various concentrations of the tested compounds and the viability of cells after treatment was determined using MTS assay. IC50 values were calculated for all tested compounds from dose‐ response curves. In [MoO2(HL)(MeOH)]+, the aroylhydrazone is coordinated to {; ; ; ; ; MoO2}; ; ; ; ; 2+ core through the O, N, O donor atoms (Figure 1). All compounds exhibited no cytotoxic activity on HepG2, whereas substantial cytotoxicity on THP‐1 cells was observed. Molybdenum(VI) compounds exhibited greater cytotoxicity than the free hydrazone.

Aldoxime derivatives of pyridinium cation are biologically active compounds having potential pharmacological application. As strong nucleophilic agents in aldoximato zwitterion form, R-py+–C(H)=N–O–, they are used as effective reactivators of the human acetylcholinesterase inhibited by organophosphorus compounds (e.g. pesticides or chemical warfare nerve agents). The biological function and pharmacological effects of pyridinium aldoximes are closely related to their structural features, acidity of aldoxime functional group and their capability to coordinate to metal ions. Aquapentacyanoferrate(II) ion, [Fe(CN)5(OH2)]3–, represents a structural model of important octahedrally coordinated iron(II) centres found in protein macromolecules. The labile sixth coordination site and great stability of pentacyanoferrate(II) moiety in aquapentacyanoferrate(II) ion make it a good selective probe for the investigation of ambidentate ligand reactivity and the coordination mode. N, N'-bis(pyridinium-2- aldoxime)tetramethylene dibromide (QMB2-2Br) and its 3-isomer (QMB3-2Br) and 4-isomer (QMB4-2Br) were used to explore the influence of the position of aldoxime group on pyridinium ring on their properties and reactivity towards [Fe(CN)5(OH2)]3–. Ionization ability in the order QMB2 > QMB4 > QMB3, determined in aqueous media, was found to be in good agreement with their structural differences and consequent resonance stabilization of their aldoximato zwitterions. The kinetic study on the formation and dissociation of the corresponding [Fe(CN)5(QMB)](3–n)– complexes was performed in buffered aqueous media in pH range 4–11 at 25 °C and the deduced kinetic parameters have been correlated with - and - bonding properties of the examined bis(pyridinium aldoxime) ligands. The notable influence of the ligands' isomerism on their reactivity towards aquapentacyanoferrate(II) ion was obtained. The established decrease of the formation rate constants in the order QMB3 > QMB4 > QMB2 was in accordance with the increasing steric hindrance. The slightly pH-dependent dissociation rates were found, which along with the distinctive position of the MLCT band in the electronic absorption spectra, strongly supported the N-coordination of the aldoxime group. Furthermore, all complexes can be classified as labile while the ligands have more pronounced σ-donor than π-acceptor ability. Spectroscopic, elemental and thermogravimetric analysis of the isolated solids support the fact that all complexes are mononuclear pentacyano(QMB)ferrates(II).

Aim: Leukoplakia is the most common potentially malignant oral disorder (PMOD) with a worldwide prevalence of 2% and a malignant transformation rate of 0.3%. Squamous cell carcinoma antigens (SCCA) are part of the serpins family. They are involved in inflammation, apoptosis, cell- migration and are expressed in the squamous cell epithelium. SCCA2 protein is 92% homologous to SCCA1, however these antigens are not expressed concurrently and play different roles in the human body. Their role in the healthy and malignant tissues is not clearly understood. Materials and methods: Unstimulated whole saliva (UWS) and stimulated whole saliva (SWS) were sampled from 5 patients with leukoplakia (5 male) and 12 age and sex matched control subjects. Leukoplakia was diagnosed with a C2certainty factor. Sandwich human SCCA1 and SCCA2 ELISA Kits, My BioSource, San Diego, USA, were used. Respondents’ alcohol consumption, papilla bleeding index (PBI), smoking, drug consumption and medical condition was registered. This study has been funded by the Croatian Science Foundation (IP-09-2014- 9376). Results: Salivary SCCA1 was significantly higher in patients with leukoplakia in UWS (U=4, p=0.045). No statistically significant difference in SCCA2 levels in UWS and SWS between the two groups was found. Due to the small sample size, smoking, PBI, medical conditions, drug and alcohol consumption had no statistical significance. Conclusion: Salivary SCCA1 could serve as a satisfactory biomarker for leukoplakia. However, a greater sample is needed to establish the diagnostic value of the investigated molecules. To our knowledge, SCCA1 and SCCA2 have not yet been measured in patients with leucoplakia.

Exactly a hundred years ago the Department of Chemistry and Biochemistry was established within the newly founded School of Medicine in Zagreb. Its founder, first Department Head and Professor was Fran Bubanović. Education of future physicians in medical chemistry and biochemistry is continuously performed since 1918. This paper aims to provide an insight in teaching activities at the Department throughout almost forty years of Bubanović’s university career. The contribution of professor Bubanović is unmeasurable not only in the early years of the Department teaching activity, but also to the organisation and management of courses and the Department throughout many years, as well as to university literature in chemistry. We believe that professor Bubanović has made a profound impact on many generations of students with his dedicated work, and us, his successors, he left forever indebted.

Prikazuju se rezultati pilot istraživanja, prvi put se ovi markeri određuju u slini. Rezultati upućuju da se radi o vrijednom markeru za rano otkrivanje oralnoga karcinoma.

Prije točno sto godina, u sklopu tek osnovanog Medicinskog fakulteta u Zagrebu, utemeljen je Zavod za kemiju i biokemiju. Njegov utemeljitelj, prvi predstojnik kao i nastavnik bio je profesor Fran Bubanović. Od te davne 1918. godine unutar Zavoda se kontinuirano provodi naobrazba budućih liječnika iz medicinske kemije i biokemije. Ovim radom želimo dati uvid u nastavnu aktivnost koja se odvijala na Zavodu kroz četrdesetak godina Bubanovićeve sveučilišne karijere. Neizmjeran je doprinos profesora Bubanovića načinjen ne samo u ranom početku nastavne aktivnosti Zavoda, nego i dugogodišnjoj organizaciji i vođenju nastave i Zavoda, kao i u području fakultetske stručne i nastavne literature iz područja kemije. Smatramo da je profesor Bubanović svojim predanim radom ostavio traga na mnogim generacijama svojih studenata, a nas, njegove nasljednike na Zavodu neizmjerno zadužio.

Enaminones as a class of compounds containing conjugated N–C=C–C=O system represent a promising group of potential therapeutics showing anticonvulsant, antimicrobial, antioxidant, antitumor and cytogenetic biological activity. Due to their lipophilicity, sensitivity to minor structural changes in the N–C=C–C=O backbone and stability in solution under physiological conditions they are used as suitable starting material for the synthesis of bioactive molecules in the field of medicinal chemistry. These tridentate ONO donors also represent a significant group of ligands in the coordination chemistry of transition metals. Trying to give more insight into the correlation between chemical structure and biological activity, six non-symmetric acyclic enaminones, 4-[(2-hydroxy-5- methylphenyl)amino]pent-3-en-2-one (H2L1), 4-[(2- hydroxy-4-methylphenyl)amino]pent-3-en-2-one (H2L2), 4-[(4-hydroxy-2-methylphenyl)amino)]pent- 3-en-2-one (H2L3), 3-[(2-hydroxy-5- methylphenyl)amino]-1-phenylbut-2-en-1-one (H2L4), 3-[(2-hydroxy-4-methylphenyl)amino]-1-phenylbut-2- en-1-one (H2L5) and 3-[(4-hydroxy-2- methylphenyl)amino]-1-phenylbut-2-en-1-one (H2L6), have been synthesized and characterized (Fig. 1). The cytotoxicity of enaminones was investigated against THP-1 and HepG2 cells in vitro and corresponding IC50 values were determined by MTS assay. The antibacterial activity was tested by microdilution method against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Moraxella catarrhalis bacterial strains to assess their MIC values. To investigate the coordination ability and influence of metal ion complexation on cytotoxicity and antibacterial activity of prepared enaminones, twelve molybdenum(VI) complexes of different nuclearity containing enaminone ligands H2L4 or H2L5 were prepared. The obtained complexes, [MoO2(L5 or 4) (MeOH)]MeOH (1MeOH and 2MeOH), [MoO2(L5 or 4) (D)] [D = pyridine, (1a and 2a), 3-methylpyridine, (1b and 2b) and 4-methylpyridine, (1c and 2c)], [{; ; ; ; ; MoO2(L5 or 4)}; ; ; ; ; 2(D)] [D = 4, 4'- bipyridine (1d and 2d)], and complexes [MoO2(L5 or 4)]n (3 and 4) were characterized and tested for cytotoxic and antibacterial activities. Enaminones and their Mo(VI) complexes were characterized by thermal analysis, IR spectroscopy and X-ray diffraction.

According to the World Health Organization, oral cancer (OC) is the eighth most common cancer in the world, with a five-year survival rate of 50%. Squamous cell carcinoma antigens (SCCA) are part of the family of inhibitory serine protease inhibitors (serpins), involved in inflammation, apoptosis, cell migration and invasiveness and expressed in the squamous cell epithelium. SCCA2 protein is 92% homologous to SCCA1, however these antigens are not expressed concurrently and play different roles in the human body. Apart from inactivation of certain enzymes, their role in the healthy and malignant tissues is not clearly understood. The origin of SCCA antigens in serum and body fluids remains unclear. Unstimulated whole saliva (UWS) and stimulated whole saliva (SWS) was sampled from 10 patients with OC (2 female, 8 male) and 15 control subjects (6 female, 9 male). Only patients with histologically diagnosed oral squamous cell carcinoma were included in the study. Sandwich human SCCA1 and SCCA2 ELISA Kits, My BioSource, San Diego, USA, were used. Respondents’ alcohol consumption, papilla bleeding index (PBI), smoking, drug consumption and medical condition was registered. This research was funded by the Croatian Science Foundation (IP-2014-3796). Salivary SCCA1 was significantly higher in OC patients, both in UWS and SWS (Mann Whitney U test, U = 14, P = 0.0004). Although no statistically significant difference in SCCA2 levels in UWS and SWS between the OC and control group was found (U = 43, P = 0.081), the average value in the control group was 895.92 pg/ml, while in OC patients 319, 45 pg/ml. Due to the small sample size, smoking, PBI, medical conditions, drug and alcohol consumption had no statistical significance. Salivary SCCA1 could serve as a satisfactory biomarker for OC. However, a greater sample is needed to establish the diagnostic value of the investigated biomarkers. To our knowledge, SCCA1 and SCCA2 have not yet been measured in the saliva of OC patients.

Background: Oral squamous cell carcinoma (OC) is one of the ten most frequent cancers in the world, with a 5-year survival rate between 30- 60%. Melatonin (MLT) has anti-oxidant, anti- inflammatory and immunomodulating properties and is considered to contribute in protecting the oral cavity from tissue damage caused by endogenic and exogenic factors. Squamous cell carcinoma antigen (SCCA) has been recognized as a potential serum marker for various squamous cell carcinomas and other diseases. However, most of the studies on serum SCCA and OC have not distinguished SCCA1 and SCCA2, due to their similar chemical structure. It is important to mention that the two molecules play different roles in the organism. Aim of the proposed study is to determine salivary MLT and SCCA1 concentrations in patients with OC. Method: Unstimulated whole saliva (UWS) and stimulated whole saliva (SWS) was sampled from 29 patients with OC (21 male, 8 female) and 29 age and sex matched healthy subjects (22 male, 7 female). Only patients with histologically diagnosed oral squamous cell carcinoma were included in the study. Sandwich human SCCA1 ELISA kits (My BioSource, San Diego, USA) and Melatonin direct saliva ELISA kits (IBL International GmbH, Hamburg, Germany) were used. This research has been funded by the Croatian Science Foundation (IP-09-2014-9376). Results: Melatonin UWS levels were higher in the OC than in the control group (p=0.045 ; median OC: 2.18 pg/mL(95% CI:1.37- 2.88), median control group: 0.84 pg/mL(95% CI: 0.50-1.71)). SCCA1 levels were higher in the OC group both in WUS (p=0.030 ; median OC: 244, 26 pg/mL(95% CI:133, 00 to 513, 27), median control group: 2pg/mL(95% CI: 2, 00-321, 47)) and WSS (p=0.014 ; median OC: 478, 03 pg/mL(95% CI:125, 16-618, 74), median control group: 2, 00(95% CI:2, 00- 207, 46)). Conclusion: Salivary MLT and SCCA1 could serve as satisfactory biomarkers for OC. To our knowledge, MLT and SCCA1 have not yet been measured in the saliva of OC patients. Disclosure: Funded by Croatian Science Foundation (IP-09-2014- 9376)

In recent years, mechanochemistry is attracting interest and sympathy of many scientist [1]. In general, it is promoted by hand grinding or ball milling [2]. Ball milling is an efficient method, commonly used both in organic and inorganic chemistry. Hydrazones are class of as chelating agents, used in coordination chemistry and their metal complexes are of a great importance. The significance of studying hydrazones arises from the fact that they have increased biological activity and they form particular materials with unique properties [3, 4]. In particular are known as antioxidants, vasidilatators, agents again tuberculosis and tumors, ect. [5, 6] Moreover, molybdenum complexes with hydrazone ligands have been reported to possess interesting antibacterial activities. The presented research considers green protocol and highly efficient synthesis of series of aromatic hydrazones. [7]. Ball milling was deliberated as solventless method promoting eco-friendly reaction conditions. The ligands were prepared by the condensation of salicylaldehyde derivative (A1-3) and appropriate hydrazide (isoniaside or nicotinic acid hydrazide, H1-3), as shown in the Scheme 1. Solid state preparative procedures were also followed by conventional synthetic procedure, in solution, and different forms of ligands were isolated (polymorphs and solvates), whose molecular and crystal structure was determined by the single crystal diffraction method. Molybdenum(VI) mononuclear and dinuclear complex with the obtained ligands were synthesized and characterized. The prepared compounds were biologically tested. With the targeted synthetic policy and recognized correlation of structure and activity, biologically active compounds can be evolved and expanded.

Uromodulin je pleiotropni glikoprotein s brojnim fiziološkim funkcijama. Do sada je poznata njegova regulacijska uloga u transportu soli, protektivna uloga mokraćnog sustava od infekcija kao i bubrežnih kamenaca, te bitna protektivna uloga u oštećenju bubrežne funkcije te i samog urođenog imuniteta. Najnoviji podaci predstavljaju niski serumski uromodulin kao nezavisni čimbenik rizika za kardiovaskularnu smrtnost. Osim navedenog, polimorfizam gena UMOD rs13333226, tj. varijantni alele G se povezuje s nižim vrijednostima arterijskog tlaka (AT). S obzirom na navedeno cilj navedenog rada bio je ispitati ulogu varijabilnosti gena UMOD u prehipertenziji (PHT), kao potencijalnog biljega tranzicije PHT u hipertenziju. Ispitivanje je obuhvatilo 496 ispitanika oba spola, 157 s AT < 120/80 mmHg) i 339 s AT ≥ 120/80 i 45 godina. Ustanovljena povezanost između varijantnog alela i nižih vrijednosti AT nije statistički značajna, što se može objasniti malim kontrolnim uzorkom. Nadalje, rezultati analize povezanosti genotipova UMOD s ostalim ispitivanim parametrima pokazali su povezanost varijantnog alela G alela s nižim vrijednostima ukupnog kolesterola, LDL- kolesterola i glukoze u obje dobne skupine. Detektirana je statistički značajna povezanost heterozigotnog genotipa A/G s visokim vrijednostima LDLkolesterola. Ovaj nalaz pozitivne heteroze otvara daljnja pitanja uloge kako i samog uromodulina tako i genotipa UMOD u metabolizmu lipida i u metabolizmu glukoze. UMOD predstavlja pleitropni gen koji ima svoju ulogu u razmatranju kardiorenalnog kontinuuma.

Drugo izdanje priručnika daje sažeti pregled doprinosa molekularne biologije u području genetike i epigenetike, genomike, transkriptomike, proteomike, nutrigenomike i farmakogenomike, te prikaz modernih trendova u primjeni najnovijih dostignuća u kliničkim i znanstvenim laboratorijima.

Introduction: Statins have been shown to possess antioxidant effects and our previous results showed that simvastatin significantly decrease malondialdehyde (MDA) levels in plasma and tissue of rats. Here we wanted to investigate the influence of rosuvastatin (ROSU) on MDA level and superoxide dismutase (SOD) activity in plasma and liver of rats, both considered to be markers of oxidative stress. Material and Methods: Thirty-six male Wistar rats were divided in one control group (saline) and two experimental groups (ROSU 5 mg/kg/day and 10 mg/kg/day). Drug and saline were given orally for period of 3 weeks. Animals were sacrificed with ether 24 hours after last dose. Blood samples were obtained directly from heart and liver tissue was rinsed with saline. MDA in plasma was measured with high performance liquid chromatography using commercially HPLC kits, while in liver using Drury method. SOD activity was measured in plasma with sensitive spectrophotometric assay and in liver using Flohé and Ötting method. Data were analyzed using ANOVA test. Results are expressed as the means and SDs. Results: ROSU in both doses decreased MDA level in liver but decrease was significant (p

Statins are drugs widely used to treat hypercholesterolemia and reduce the risk of cardiovascular morbidity and mortality in patients with coronary heart disease. The mechanism of their action may be the inhibition of oxidant formation and blocking the effects of reactive oxygen species (ROS). As reported in recent studies, rosuvastatin has a beneficial effect on the plasma lipid profile and can reduce serum cholesterol and ROS formation in diabetic patients. The aim of this work was to determine the influence of rosuvastatin on the prooxidant- antioxidant balance in rat kidneys. This study was performed on male normolipidemic Wistar rats (N=36) divided into two control (treated with saline 5 mg kg-1 per day) and two experimental rosuvastatin-treated (5 and 10 mg kg-1 per day) groups. Animals were treated orally for 21 days. Malondialdehyde (MDA) – parameter of lipid peroxidation was measured in the kidney tissue homogenate of control and treated animals using the method by Drury et al. (1997). Data were analysed using t-test. Results are expressed as Mean±STD. MDA concentration was significantly lower in both experimental groups compared to controls: 0.41±0.05 vs. 0.62±0.08 µmol g-1 tissue (5 mg kg-1 per day vs. control ; p

There are diverse experimental data about the influence of gemfibrozil (GEM) on the production of hydrogen per- oxide (H2O2) and antioxidant enzymes. We investigated the influence of GEM treatment on the production of malondialde- hyde (MDA) level in tissues of normolipidaemic Wistar and Fisher rats which is an index of lipid peroxidation. Because serum paraoxonase 1 (PON1) is an important enzyme with specific protective function on metabolism of lipid peroxides, we examined the influence of GEM on PON1 activity in liver and serum. MDA level and enzyme activities were also determined 10 days after withdrawal of GEM treatment. The significantly increased levels of MDA in liver, kidney and heart of both rat strains were obtained after 3 weeks of GEM treatment. We propose two possibilities for the increase of MDA levels caused by GEM, induction of peroxisome proliferation and activities of enzymes that participated in occurrence of H2O2 and possible reduction of enzyme activities including in H2O2 metabolism. Ten days after withdrawal of GEM treatment, MDA levels in all tissue levels of both rat strains were less in comparison with GEM treatment. GEM caused a significant drop of PON1 activity in serum and liver of Fisher rats, and in liver of Wistar rats. We suggest that GEM, through induction of lipid peroxi- dation, caused the damage of hepatocytes with consequent reduction of PON1 synthesis. The increase in PON1 activity in serum and tissues of both rat strains 10 days after withdrawal of GEM treatment shows the fast recovery of enzyme synthesis.

Izbor i omjer makronutrijenata u prehrani je uz druge čimbenike i genetički determiniran te je predmet istraživanja nutrigenetike. AT1R je dominantan receptor u kardiovaskularnom sustavu za regulaciju hemodinamskih procesa i niza drugih procesa važnih u patofiziologiji debljine i metaboličkog sindroma (MetS). Cilj studije bio je ispitati interakcije između polimorfizma gena za angiotenzinski receptor AT1R i vrste prehrane te utvrditi njihovu povezanost s metaboličkim sindromom (MetS) i njegovim sastavnicama. Istraživanje je uključivalo 527 ispitanika (265 s MetS i 262 kontrolna ispitanika). Određene su antropometrijske osobine (visina, težina, opseg struka, arterijski tlak) i biokemijski parametri (trigliceridi, ukupni-, LDL- i HDL-kolesterol i glukoza) te je analiziran tip prehrane. Mediteranska prehrana je uključivala maslinovo ulje, voće i povrće, ribu, bijelo meso i orašaste plodove barem tri puta tjedno. Kontinentalni tip prehrane se sastojao od crvenog mesa i mesnih prerađevina, manjih količina voća i povrća, rijetke konzumacije ribe, bez maslinovog ulja. Genotipizacija AT1R 1166A>C provedena je metodom PCR-RFLP. Rezultati: Ispitanici koji su preferirali kontinentalni tip prehrane imali su gotovo četiri puta veće izglede za razvoj pretilosti (OR=3, 88 ; 95% CI: 1, 12-13, 36 ; p=0, 032) i značajno veći rizik za razvoj MetS (OR=13, 86 ; 95% CI: 5, 69-33, 78 ; p

Aetiology of metabolic syndrome (MetS) is complex and involves environmental and genetic factors and their interactions. Because of the important role of apoE in lipid metabolism and adipocyte activity, APOE might serve as a potential determinant of MetS. In this study of 149 subjects, aged 20-35 years, we determined whether the variable forms of functional gene APOE in interaction with environmental factors (nutrition) represent a potential genetic risk factors for increased susceptibility to MetS. Biochemical and anthropometric parameters were measured by standard laboratory methods. The APOE was genotyped by real-time PCR. No APOE genotype significantly increased the risk for MetS. Significant association was found between APOE polymorphism with elevated blood pressure (p=0.019). APOE alleles were significantly associated with the concentration of TC and LDL-C (p=0.002 and p

Background: It was suggested that statins exerted their protective effect against Alzheimer disease (AD) due to inhibition of cholinesterase (ChE) activity. Results of some nonclinical and clinical studies have shown that statins either do not influence or decrease enzyme catalytic activity. Our previous studies showed that simvastatin increased plasma and liver ChE activity in normolipidemic rats. In respect to these findings we studied influence of simvastatin (SIMV) and fenofibrate (FENO) on ChE activity in hyperlipidemic rats. Methods: Twenty-one male hyperlipidemic Zucker rats were divided into one control and two experimental groups. One experimental group was on SIMV treatment (50 mg/kg/day) and another on FENO treatment (30 mg/kg/day). Both agents were given orally for 3 weeks, as well as saline for control group. Animals were sacrificed 24 hours after the last dose. Blood samples were obtained directly from heart. Liver and brain tissue were rinsed with saline. ChE activity was determined by Ellman’s spectrophotometric method using butyrylthiocholine and acetylthiocholine as substrates. In liver tissue the ChE activity was measured with and without ethopropazine hydrochloride. Enzyme activities were expressed as μmol of substrate hydrolysed per min per ml of serum or g of tissue. Data were analyzed using ANOVA and Dunnett's test. Results were considered as significant with p

Peroxisome proliferator activated receptor-gamma (PPARG) and lipoprotein lipase (LPL) play important role in lipid homeostasis, insulin resistance and adipogenesis, and their gene variability could be considered as predictive genetic markers for metabolic syndrome (MetSy). The aim of the study was to estimate possible associations of PPARG (Pro12Ala) and LPL PvuII (+/-) polymorphisms with MetSy and its traits. Study included 527 subjects. According to the modified National Cholesterol Education Program Adult Treatment Panel III definitions, subjects were classified into the metabolic syndrome group and control group. Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism methods. In the total sample, LPL variants were associated with waist circumference (chi2 = 7.263, d.f = 2, p = 0.026) and with BMI (chi2 = 6.549, d.f = 2, p = 0.038), where PvuII (+/+) genotype carriers had the highest risk for increased waist circumference (specific PvuII (+/+) vs. others analysis chi2 = 7.033, p = 0.008) and increased BMI (specific PvuII( +/+) vs. others analysis chi2 = 5.154, p = 0.023). LPL gene variants were also associated with HDL-C levels (chi2 = 6.901, d.f = 2, p = 0.032), where PvuII (-/-) genotype carriers had higher HDL-C values in comparison to others (specific Pvu (+/+) vs. others analysis chi2 = 6.504, p = 0.011). Furthermore, PvuII (-) allele carriers had significantly lower glucose (allele based analysis Add Value = -0.0878, chi2 = 5.878, d.f. = 1, p = 0.015). Significant interaction was detected between PPARG and LPL that affected HDL-C levels in male population (chi2 = 11.790, d.f = 1, p = 0.0006) in the manner that Ala/PvuII(+) contributed to the lowest HDL-C values (Specific Ala/ Pvu(+) vs. others analysis was chi2 = 11.750, p = 0.0006). According to obtained results LPL and PPARG gene variants could be susceptibility factors of obesity and lipid status, contributing to development of MetSy, particularly in males. Because of antiatherogenic function of HDL-C, the identification of genetic variants associated with HDL-C can provide useful information related to genotype-phenotype relationships. Since the interplay between PPARG and LPL gene and gender seems to be significant it could point to the personalized behavioural recommendations for prevention of metabolic and cardiovascular diseases.

Introduction: Pyridinium aldoximes are known as pharmacologically important, strong nucleophilic agents that are effective antidotes against poisoning by organophosphorus compounds (pesticides, chemical warfare nerve agents, drugs used in treatment of cholinergic disorders). Furthermore, the biological function of aldoximes and the mechanism of their metabolism are related to their chelation to metal ions. Nowadays, the aldoxime-induced reactivation of the phosphorylated and thus inhibited acetylcholinesterase (AChE, E.C.3.1.1.7) is the primary therapeutic approach to organophosphorus poisoning. The most commonly pyridinium aldoxime used in routine human therapy is N-methylpyridinium-2-aldoxime chloride (PAM2-Cl) known as Pralidoxime Chloride. Its structural and chemical properties in aqueous media have been correlated with the N-methylpyridinium-3-aldoxime iodide (PAM3-I) and N-methylpyridinium-4-aldoxime iodide (PAM4-I). Subsequently, their coordination ability toward the aquapentacyanoferrate(II) ion, [Fe(CN)5(OH2)]3–, as a model of biologically important macromolecules with the labile sixth coordination site, has been elucidated. Materials and methods: PAM3-I and PAM4-I were synthesized, while PAM2-Cl (Sigma-Aldrich) was reagent-grade and used as purchased. Na3[Fe(CN)5(NH3)]•3H2O (Sigma-Aldrich) was used as a precursor of [Fe(CN)5(OH2)]3–. The pH of the reaction mixtures was adjusted using Britton-Robinson buffers. All kinetic and thermodynamic measurements were carried out on Varian Cary Bio 100 spectrophotometer. The 1H and 13C NMR spectra were recorded at ambient temperature in DMSO-d6 on Bruker Avance 600 spectrometer. FT-IR spectra were recorded on a Perkin Elmer Spectrum GX, Series R spectrometer. Results: Predominant ionic structures of the investigated pyridinum aldoximes in aqueous solution (pH range 4.0–11.5) as well as their ionization ability have been established. The electronic, vibrational and NMR spectral characterization as well as nucleophilic properties of the investigated aldoximes are found to be in agreement with the bond distances and valence angles obtained from the previously performed X-ray structure analysis. The thermodynamic and kinetic properties of their reactive forms toward the iron(II) center in the pentacyanoferrate(II) moiety are evaluated. Conclusion: The nucleophilic strength of the investigated aldoximes at physiological pH is found to be in the following order: PAM2-ClPAM4-IPAM3-I. The pyridinium aldoxime pentacyanoferrate(II) complexes can be classified as thermodynamically instable and kinetically labile.

PPARG – receptor za aktivator proliferacije peroksisoma tip gama je nuklearni transkripcijski faktor, koji regulira ekspresiju niza gena s važnom ulogom u metabolizmu lipida i glukoze, transportu masnih kiselina, diferencijaciji adipocita i upali. Polimorfna varijanta PPARG Pro12Ala modulira transkripcijsku aktivnost, a time i stimulaciju ciljnih gena među koje pripadaju i LPL, IL-6, ACE i AT1R. Stoga funkcija PPARG može predstavljati ključnu ulogu u razvoju dislipidemije, debljine, hiperglikemije i hipertenzije, važnih rizičnih čimbenika za razvoj metaboličkog sindroma (MetS).

Biological importance of pyridinum aldoximes is closely related to their structure, acidic properties, nuchleophilicity and coordination ability. Compounds of pyridinium aldoxime type have been shown to exhibit a great variety of pharmacological effects. An important pharmacological application is derived from their capability to reactivate organophosphate-inactivated acethylcholinesterase. N-methylpyridinium-2-aldoxime chloride (PAM2-Cl), N-methylpyridinium-3-aldoxime iodide (PAM3-I) and N-methylpyridinium-4-aldoxime iodide (PAM4-I) were used to explore the influence of the position of the aldoxime group in N-methylpyridinium ring on the structure, chemical properties and reactivity toward the pentacyanoferrate(II) moiety. Aquapentacyanoferrate(II) ion, an useful model for biological macromolecules with a labile sixth coordination site, was used as a metal reagent. Crystal structure characterization was done for PAM3-I and compared with the known structures of PAM4-I and PAM2-Cl. Ionization ability for all three ligands was determined and was found to be in good agreement with their structural differences and consequent resonance stabilization of their aldoximato forms. Reactions of the examined ligands with the aquapentacyanoferrate(II) ion were studied in buffered aqueous media by UV/Vis spectroscopy. Comparative kinetic and equilibrium studies on the formation and dissociation of the produced pentacyano(ligand)ferrate(II) complexes were carried out at 25°C and I=0.1 M. Equilibrium constants and stoichiometry of the complexes at pH 6 were determined by mole ratio method. The kinetic properties of the produced complexes were determined in a pH range 5.0-11.5 and correlated with σ-donor and π-acceptor capability of the ligands. Furthermore, the pH-dependency of the complex dissociation rate was analyzed in terms of ionization ability of the coordinated ligand. The obtained values for the equilibrium constants, identified as apparent formation constants, along with the kinetic parameters suggested that all three complexes can be classified as instable and labile.

Atherosclerosis and cardiocerebrovascular diseases are the leading cuases of morbidity and mortality in developed countries where a high number of such patients are daily admitted in intensive care and emergency medicine departments. Most diseases have a genetic component ; each individual possesses a genome that involves a certain risk disease.

Background: Adiponectin is an adipose tissue-derived adipokine linked to central obesity and ADIPOQ variants are promising markers for understanding the genetic base of obesity. We aimed to evaluate the relation of adiponectin concentrations and ADIPOQ gene variants to abdominal obesity and hypertension in young subjects. In addition, influence of the gene-environment (diet) interaction was estimated.

Background: 5-Hydroxytryptamine (5-HT, serotonin) plays an important role in the central nervous control of energy balance and is involved in several biological processes including mood, appetite, sleep, libido, memory, and body weight regulation. Brain-derived neurotrophic factor (BDNF) is also currently recognized as an important participant in the regulation of food intake. The aim of this study was to evaluate whether the 5-HTTLPR S/L and BDNF Val66Met gene variants are associated with obesity in a sample of adults of Croatian origin. Materials and methods: 462 individuals were investigated including 301 obese (BMI ≥ 30 kg/m2) and 161 lean (BMI < 25 kg/m2) (mean age ± SD was 49 ± 8 years). Genotyping of triallelic structure of 5-HTTLPR (LA, LG, S) and of BDNF Val66Met polymorphisms was performed using the RealTime based allele specific PCR methods. Results: In the whole group we found no associations between 5-HTTLPR S/L and BDNF Val66Met polymorphisms and obesity. When we compared male and female samples, we observed statistically significant differences in the distribution of 5-HTTLPR genotypes: 5-HTTLPR LALA genotype was more frequent in the group of lean women comparing to the group of lean men (41% and 28% respectively, P = 0.022). The 5-HTTLPR S and BDNF Met allele carriers have higher risk to develop obesity (OR 2.07) than non carriers (P = 0.038). Discussion: Our findings indicate that 5-HTTLPR polymorphism may be linked with obesity in adult female population, reinforcing the role of the serotonin transporter as a risk factor for the obesity phenotype. SERTPR and BDNF gene interactions could additionally predispose to obesity risk.

Background. Metabolic syndrome (MS) is a cluster of modifiable risk factors including hypertension, abdominal obesity, dyslipidemia and insulin resistance, associated with nonmodifiable risk factors, such as age, sex and genetic background. We investigated the possible role of gene polymorphisms of PPARG (Pro12Ala), ApoE (ε2, ε3, ε4), LPL (P+/-), IL-6 (-174G>C), ACE (I/D) and AT1R (1166A>C) in MS. Methods. 516 individuals were investigated including 263 patients with MS and 253 subjects without MS criteria. Genotyping was performed using PCR based methods. Results. In female group associations were found for: LPL and ACE with MS (p=0.04) ; PPARG and LPL with blood pressure, (p=0.04) ; LPL with cholesterol and LDL (p=0.01 and p=0.05, respectively). Significant gene interactions observed between: APOE and PPARG, ACE and APOE were associated with BMI (p=0.01 and p=0.05, respectively) ; LPL and PPARG were associated with triglycerides (p=0.03). For males we found associations of: LPL variants with MS (p=0.02), BMI (p=0.002) and waist circumference (p=0.008) ; PPARG and APOE with BMI (p=0.05) ; IL-6 with CRP (p=0.02). Significant gene interactions observed between: PPARG and AT1R were associated with blood pressure (p=0.05) ; PPARG and APOE with triglycerides (p=0.02) ; PPARG and APOE, PPARG and IL6 (p=0.03), ACE and APOE (p=0.0002) with cholesterol ; PPARG and LPL (p=0.003), PPARG and IL6 (p=0.06) with HDL ; PPARG and IL6 (p=0.01), ACE and APOE (p=0.04) ; PPARG and APOE, LPL and ACE (p=0.01), AT1R and ACE (p=0.06) with CRP. Conclusion. Gene variants of PPARG, APOE, LPL, ACE, AT1R and IL-6 could be susceptibility factors of obesity, lipid status, and glucose intolerance.

There are diverse experimental data about the influence of gemfibrozil (GEM) on the production of hydrogen peroxide (H(2)O(2)) and antioxidant enzymes. We investigated the influence of GEM treatment on the production of malondialdehyde (MDA) level in tissues of normolipidaemic Wistar and Fisher rats which is an index of lipid peroxidation. Because serum paraoxonase 1 (PON1) is an important enzyme with specific protective function on metabolism of lipid peroxides, we examined the influence of GEM on PON1 activity in liver and serum. MDA level and enzyme activities were also determined 10 days after withdrawal of GEM treatment. The significantly increased levels of MDA in liver, kidney and heart of both rat strains were obtained after 3 weeks of GEM treatment. We propose two possibilities for the increase of MDA levels caused by GEM, induction of peroxisome proliferation and activities of enzymes that participated in occurrence of H(2)O(2) and possible reduction of enzyme activities including in H(2)O(2) metabolism. Ten days after withdrawal of GEM treatment, MDA levels in all tissue levels of both rat strains were less in comparison with GEM treatment. GEM caused a significant drop of PON1 activity in serum and liver of Fisher rats, and in liver of Wistar rats. We suggest that GEM, through induction of lipid peroxidation, caused the damage of hepatocytes with consequent reduction of PON1 synthesis. The increase in PON1 activity in serum and tissues of both rat strains 10 days after withdrawal of GEM treatment shows the fast recovery of enzyme synthesis.

Metabolički sindrom (MS) čini grupa sastavnica povezanih s povećanim rizikom za kardiovaskularne bolesti. Sve definicije MS-a kao ključnu odrednicu ubrajaju i abdominalni suvišak masnog tkiva. Na etiologiju abdominalne pretilosti kao i MS-a utječe međudjelovanje genetičkih i okolišnih čimbenika. Cilj istraživanja je utvrditi moguću povezanost varijantnih oblika gena za adiponektin (ADIPOQ) i peroksisom-proliferator-aktivirani receptor (PPAR2) s razvojem abdominalne pretilosti, a time i MS-a kod mladih ljudi.

Background. The role of genetic factors in the development of metabolic syndrome (MS) has been widely recognized, but the contribution of genes has not yet been fully clarified. We investigated the possible role of gene polymorphisms of PPARγ (Pro12Ala), ApoE (ε2, ε3, ε4), LPL (P+/-), IL-6 (-174G>C), ACE (I/D) and AT1R (1166A>C) in MS. Methods. Genotyping of PPARγ, LPL, IL-6, AT1R and ACE was performed by PCR-RFLP, and of APOE by real-time PCR in a group of 263 patients and 180 controls. Results. We found association of LPL variants with waist circumference (p=0.02), BMI, LDL and HDL (p=0.05) ; APOE and ACE significantly corelated to cholesterol (p=0.01), D and E2 allele contributed to its increased level while APOE2 corelated to lower HDL (p=0.03) ; ACE and AT1R variants correlated with LDL cholesterol (p=0.04) ; ACE D predisposed to increased glucose level (p=0.02). Borderline associations were found between blood pressure and combination of variant alleles of PPARg and APOE (p=0.06), APOE and ACE (p=0.06) ; BMI and APOE (p=0.06). Conclusion. Gene variants of APOE, LPL, ACE, AT1R and PPARγ could be susceptibility factors of obesity, lipid status, and glucose intolerance contributing to development of MS.

Background. Metabolic syndrome (MS) is a cluster of risk factors including hypertension, abdominal obesity, dyslipidemia and hyperglycemia. The contribution of genetic factors to the development of MS has been widely recognized, but the contribution of the genes has not yet been fully clarified. We investigated the possible role of gene polymorphisms of PPAR (Pro12Ala), ApoE, ACE (I/D) and AT1R (1166A>C) in MS. Methods. Genotyping of PPARG2, AT1R and ACE was performed by PCR-RFLP, APOE by real-time PCR in group of 281 patients and 119 controls. Associations of alleles and genotypes with biological and clinical variables were performed using UNPHASED-3.0.10. Results. In comparison to females, males were older, had higher BMI and waist circumference, higher triglycerides and glucose levels and lower HDL. Males had significantly more often high blood pressure. Age accounted for the differences in glucose levels and HDL. We found significant association of AT1R variants and the developement of MS (p=0.03), with the C allele being associated with lower risk. We found association of waist circumference with: ACE variants (p=0.03), with D allele carrying the higher risk ; AT1R with A as the risk allele (p=0.05). APOE4 variant was associated with the waist circumference (p=0.05) and the levels of HDL (p=0.03), and ACE genotype was associated with glucose levels (p=0.02), with II genotype carrying the lowest risk. Conclusion. Gene variants of AT1R, ACE and ApoE could be susceptibility factors of obesity, glucose intolerance and lipid status contributing to the development of metabolic syndrome.