Jasna Lovrić

Adiponectin Level and Gene Variability Are Obesity and Metabolic Syndrome Markers in a Young Population

BACKGROUND AND AIMS Human obesity is accepted as an important risk factor for development of MetS. Adiponectin is linked to central obesity and ADIPOQ variants are promising markers for understanding the genetic base of obesity-related disorders. We performed analyses of adiponectin concentrations and ADIPOQ variants and tested their associations with obesity and MetS in young subjects of Croatian origin. METHODS Biochemical and anthropometric parameters of MetS were obtained for 149 unrelated subjects. Adiponectin levels were measured by ELISA assay. ADIPOQ -11391G>A and -11377C>G were genotyped by real-time PCR. RESULTS BMI and WC, TG and GLUC showed inverse correlation, whereas HDL-C showed a positive correlation with adiponectin concentrations. For central obesity, we found association with -11377C>G and with -11391G>A polymorphisms. ADIPOQ -11377GG and -11391GA significantly increased the risk for the development of central obesity (OR 5.57 and OR 3.37, respectively). Significant association was found between -11391A, -11377G allele and haplotype and increased TG. -11377C>G and -11391G>A variant were significantly associated with the incidence of MetS. C>G mutation at position -11377 significantly increased the risk of MetS development (OR = 2.93). Compared with the -11391G homozygotes, carriers of the A allele had a significantly increased risk for the development of MetS (OR = 3.15). The test of overall association showed a statistically significant correlation of MetS with -11377C>G and -11391G>A haplotypes (p = 0.008). CONCLUSIONS Analysis of adiponectin concentration and ADIPOQ -11391G>A and -11377C>G gene variants may be clinically meaningful for estimation of MetS risk in a young population.

INHIBITION OF ACETYLCHOLINESTERASE BY THREE NEW PYRIDINIUM COMPOUNDS AND THEIR EFFECT ON PHOSPHONYLATION OF THE ENZYME

Three new mono-pyridinium compounds were prepared: 1-phenacyl-2-methylpyridinium chloride (1), 1-benzoylethylpyridinium chloride (2) and 1-benzoylethylpyridinium-4-aldoxime chloride (3) and assayed in vitro for their inhibitory effect on human blood acetylcholinesterase (EC 3.1.1.7, AChE). All the three compounds inhibited AChE reversibly; their binding affinity for the enzyme was compared with their protective effect (PI) on AChE phosphonylation by soman and VX. Compound 1 was found to bind to both the catalytic and the allosteric (substrate inhibition) sites of the enzyme with estimated dissociation constants of 6.9 microM (Kcat) and 27 microM (Kall), respectively. Compound 2 bound to the catalytic site with Kcat = 59 microM and compound 3 only to the allosteric site with Kall = 328 microM. PI was evaluated from phosphonylation measured in the absence and in presence of the compounds applied in a concentration corresponding to their Kcat or Kall value, and was also calculated from theoretical equations deduced from the reversible inhibition of the enzyme. Compounds 1 and 3 protected the enzyme from phosphonylation by soman and VX, whereas no protection was observed in the presence of compound 2 under the same conditions. Irrespective of the binding sites to AChE, PI for compounds 1 and 3 evaluated from phosphonylation agreed with PI calculated from reversible inhibition. Compound 3 was found to be a weak reactivator of methylphosphonylated AChE with Kr = 1.1 x 10(2) L mol-1 min-1.

Reaction of 1-benzoylethylpyridinium-4-aldoxime chloride with aquapentacyanoferrate(II)

The results of a spectrophotometric investigation of the reaction of the biologically active salt 1-benzoylethylpyridinium-4-aldoxime chloride with aquapentacyanoferrate(II) ion are presented. In spite of the presence of two donor sites, only the carbonyl group of the ligand coordinates to the iron centre. Reaction kinetics are consistent with a dissociative mechanism.

Interaction of genetic risk factors confers increased risk for metabolic syndrome: the role of peroxisome proliferator-activated receptor γ.

AIM The aim of the study was to estimate the influence of interactions between peroxisome proliferator-activated receptor γ (PPARγ) and target genes lipoprotein lipase (LPL), interleukin 6 (IL6), angiotensin converting enzyme (ACE), and angiotensin II type 1 receptor (AT1R) on metabolic syndrome (MetSy) and its traits. METHODS The study included 527 participants (263 with MetSy and 264 controls). Genotyping of PPARγ Pro12Ala, LPL PvuII (-/+), IL6 -174G>C, ACE I/D and AT1R 1166A>C was performed using polymerase chain reaction-restriction fragment length polymorphism-based methods. RESULTS Interaction between PPARγ Pro12Ala and LPL Pvu(-/+) improved prediction of MetSy over and above prediction based on a model containing no interactions (χ(2)=7.22; df=1; p=0.007). In the group of participants with PPARγ Pro12Ala or Ala12Ala genotypes, those with the LPL Pvu (-/+) or (+/+) genotype had greater odds for MetSy (odds ratio OR=5.98; 95% confidence interval CI: 1.46-24.47, p=0.013). Interaction between PPARγ Pro12Ala and IL6 -174G>C improved prediction of high fasting blood glucose (χ(2)=13.99; df=1; p<0.001). PPARγ Ala12 variant was found protective in patients with IL6 -174GG genotype (OR=0.10; 95% CI: 0.02-0.57, p=0.01), while in the case of IL6 -174C allele carriers, for PPARγ Ala12 carriers, larger odds for high glucose levels compared with Pro12 variant were observed (OR=2.39; 95% CI: 1.11-5.17, p=0.026). Interactions of PPARγ and ACE were significant for BMI. In the group with ACE DD genotype, those with PPARγ Pro12Ala or Ala12Ala genotype have greater odds for obesity (OR=9.98; 95% CI: 1.18-84.14, p=0.034). CONCLUSIONS PPARγ gene variants can, in interaction with some of its target genes, modulate physiological processes leading to the development of MetSy.

Copper(II) complexes with benzhydrazone-related ligands: synthesis, structural studies and cytotoxicity assay

The dinuclear copper(II) complexes [Cu2(LH)2] (1), [Cu2(L3OMe)2] (2), [Cu2(L4OMe)2] (3) as well as the tetranuclear complex [Cu4(L3OMe)4]·4MeOH·2H2O (2a·4MeOH·2H2O), (where LH = 2-oxybenzaldehyde benzhydrazonato, L3OMe = 3-methoxy-2-oxybenzaldehyde benzhydrazonato or L4OMe = 4-methoxy-2-oxybenzaldehyde benzhydrazonato) were synthesized by reacting copper(II) acetate with the corresponding aroylhydrazone. Reactions in the presence of N-donor ligands D (pyridine, 4,4′-bipyridine or 1,10-phenanthroline) afforded complexes with different nuclearities and Cu : L : D ratios. They can be classified as dinuclear, [Cu2(LH)2(py)2] (1py), [Cu2(L3OMe)2(py)2]·3H2O (2py·3H2O), [Cu2(L4OMe)2(py)2] (3py), [{Cu(LH)}2(bpy)] (1bpy), [{Cu(L3OMe)}2(bpy)] (2bpy), and [{Cu(L4OMe)}2(bpy)] (3bpy), mononuclear [Cu(LH)(phen)] (1phen), [Cu(L3OMe)(phen)] (2phen) and [Cu(L4OMe)(phen)] (3phen), and polynuclear [{Cu(LH)}2(bpy)]n·2nMeOH (1bpy*·2MeOH) complexes. Conversion of complexes 1py–3py to complexes 1–3 proceeded via solid state reactions. The crystal and molecular structures of 2a·4MeOH·2H2O, 2py·3H2O, 3py, 1bpy*·2MeOH, 2bpy, 1phen and 3phen were determined by the single crystal X-ray diffraction method. All complexes were characterized by means of elemental and thermogravimetric analysis, FT-IR spectroscopy and the powder X-ray diffraction method. In vitro cytotoxicity of benzhydrazone-related ligands and their copper(II) complexes against THP-1 and HepG2 cells demonstrated that the complexes are significantly more cytotoxic than the corresponding ligands.

Bis­(ethyl­enedi­amine-N,N′)(pyrimidine-2-carboxyl­ato-N1,O)­cobalt(III) bis­(tri­fluoro­methane­sulfonate) dihydrate

In the title compound, the coordination geometry of the CoIII atom is only slightly distorted from regular octahedral, and the racemic nature of the material was confirmed by X-ray structure analysis.

Spectrophotometric Studies of Reactions of the Aquapentacyanoferrate(II) Ion with Ketones. Kinetics and Mechanism of the Substitution Reaction of the Aquapentacyanoferrate(II) Ion with 1-Benzoylethylpyridinium Chloride

Abstract The carbonyl group of compounds of the phenacyl- and benzoylethyl-pyridinium type is recognized as a potential σ donor ligand which can produce complexes with the aquapentacyanoferrate(II) ion. Its reactive form is the ionized one i.e. the enolate ion. A detailed spectrophotometric investigation has been made for the reaction of the aquapentacyanoferrate(II) ion with 1-benzoylethylpyridinium chloride. The equilibrium constant of the produced complex was derived using two literatury available additional types of processing the molar ratio method’s experimental data which were not found to be applied to this type of complexes. The kinetics of this substitution reaction were studied in buffered solutions at pH = 8.0 and I = 0.05 M. The obtained results are consistent with a dissociative mechanism.

Comparative studies on conventional and solvent-free synthesis toward hydrazones: Application of PXRD and chemometric data analysis in mechanochemical reaction monitoring

Synthesis of hydrazones was performed via both conventional and solvent-free routes using the corresponding hydrazide (isonicotinic hydrazide, nicotinic hydrazide, 2-aminobenzhydrazide or 4-aminobenzhydrazide) and appropriate aldehyde (salicylaldehyde, 3-methoxysalicylaldehyde or 4-methoxysalicylaldehyde). A systematic study dedicated to solvatomorphism or polymorphism screening resulted in the formation of twelve novel crystalline forms, and eight of these were characterized via the single crystal X-ray diffraction method. In all studied structures, the molecules were assembled into endless supramolecular chains, discrete rings, chains of rings or nets. The mechanochemical synthesis employing liquid-assisted grinding was also applied and the nicotinic- and isonicotinic-based hydrazones were found to form readily from their corresponding precursors. The chemometric study using principal component analysis for mechanochemical synthesis monitoring was implemented for the first time to provide an insight into the reaction profiles. A thoughtful combination of ex situ powder X-ray diffraction and chemometric analysis was essential to identify a stepwise mechanism for the hydrazone formation via an intermediate phase. In five investigated reactions the first principal component accounted for at least 75% of the total variance, whereas in the case of two reactions this component accounted for 69.72 and 46.23% of the total variance. The hydrazones were also evaluated for cytotoxic activity in vitro. All compounds exhibited weak to moderate cytotoxicity against THP-1 and no cytotoxicity against HepG2 cells. Substantial antibacterial activity was obtained against Moraxella catarrhalis while no growth inhibition of Staphylococcus aureus, Enterococcus faecalis and Escherichia coli was observed.

Effects of simvastatin on malondialdehyde level and esterase activity in plasma and tissue of normolipidemic rats

BACKGROUND We investigated the possible non-lipid effects of simvastatin (SIMV) on paraoxonase 1 (PON1) and butyrylcholinesterase (BuChE) activity, as well as on malondialdehyde (MDA) levels in normolipidemic rats. METHODS Two experimental groups of Wistar rats (10mg/kg/day of SIMV) and two control groups (saline) underwent a 21-day treatment period (TP). On the 22nd day one experimental and one control group of rats were sacrificed. Remaining groups of animals were sacrificied on the 32nd day of the study (10-day after-treatment period (AT)). Blood samples and slices of liver, heart, kidney, and brain tissue were obtained for the measurement of PON1 and BuChE activity and levels of MDA. Data were analyzed by means of t-test for independent samples. p values≤0.05 were considered as statistically significant. RESULTS SIMV caused a significant decrease of serum and liver PON1 activity (18-24%, p≤0.05) and MDA concentrations in the plasma, heart, liver, kidney, and brain (9-40%, p≤0.05), while plasma and liver BuChE activity increased by 29% (p≤0.05) and 18%, respectively. All effects of SIMV were largely diminished following AT. The exception was MDA, which remained significantly decreased in plasma and all tissues analyzed. CONCLUSION SIMV significantly decreased PON1 activity and MDA levels and increased BuChE activity. We suggest that the decrease of MDA levels is a beneficial therapeutic effect of SIMV, for example in cardiovascular disorders, while the increase of BuChE activity, especially in brain, may be a potential adverse effect in patients with Alzheimer disease.

Is there any association of apolipoprotein E gene polymorphisms with metabolic syndrome in a young population of Croatian origin

Abstract Background: Apolipoprotein E has an important role in lipid metabolism and adipocyte activity and apo E gene (APOE) might serve as a potential determinant of metabolic syndrome (MetS). Aim: The aim of the presented study was to investigate the association between APOE polymorphism and MetS in young adult subjects of Croatian origin. Methods: This study measured biochemical and anthropometric parameters of 149 young (aged 20–33) subjects. The APOE was genotyped by real-time PCR. Results: No APOE genotype significantly increased the risk for development of MetS. Significant association was found between APOE polymorphism and elevated blood pressure (EBP) (p = .019). The carriers of the ɛ4 allele had decreased risk for EBP (OR = 0.28, 95% CI) compared to ɛ3 allele carriers (ɛ3 allele vs others, χ2 = 7.08; p = .005). APOE alleles were significantly associated with the concentration of TC and LDL-C (χ2 = 12.11, p = .002 and χ2 = 15.76, p < .001, respectively). With diet as a modification covariate there was a significant correlation of APOE alleles with the concentrations of adiponectin and leptin (χ2 = 7.076; p = .029 and χ2 = 7.46; p = .024, respectively). Conclusion: Although APOE variants were not confirmed as the risk factor for development of MetS, the APOE alleles were associated with some of the metabolic parameters in young Croatian subjects. The relation of APOE alleles with a concentration of adiponectin and leptin depends on the diet intake.