On the origin of the Slovenian Cika cattle.

In 2002, it was discovered that several Cika cattle in the mountain areas of Slovenia had escaped the official policy of cross-breeding. Here, we report a genetic characterization to assess their status as autochthonous breed. We compared genotypes for 14 microsatellite markers in 150 Cika cattle individuals with data from 16 Central European cattle breeds. We show that Cika cattle are genetically as diverse as other Eastern Alpine breeds, are more diverse than Austrian Simmental but less than the Balkan Busha cattle. STRUCTURE analysis showed Pinzgauer admixture in several individuals but also indicated a unique genetic identity for Cika. This analysis also allowed a selection of the most genetically pure Cika individuals as assessed by the panel of microsatellites. These original Cika cattle form an Eastern Alpine breed cluster together with Pinzgauer and Pustertaler cattle. Cika cattle should be considered as an authentic and valuable genetic resource, which offers clear opportunities for sustainable agriculture and landscape conservation in marginal and mountain areas.

Comparative binding effects of aspirin and anti-inflammatory Cu complex in the active site of LOX-1.

(1)H NMR Saturation Transfer Difference (STD) experiments were applied to study the binding of aspirin and of an anti-inflammatory complex of Cu(I), namely [Cu(tpp)(pmt)](2) [pmt = 2-mercaptopyrimidine), synthesized in an attempt to develop novel metallotherapeutic molecules. While aspirin showed only very weak binding, the complex [Cu(tpp)(pmt)](2) clearly favored binding to LOX-1. In silico docking experiments in LOX-1 showed that aspirin does only weakly bind to LOX-1, while the complex binds with high affinity. In addition, docking experiments and molecular dynamics (MD) simulations showed that the complex binds via hydrogen bonding (HB), to an allosteric site of LOX-1, revealing that this enzyme has more than one accessible site for complex metallotherapeutic molecules. When aspirin was added in the solution containing LOX and the complex [Cu(tpp)(pmt)](2), the former was shown to hinder the binding of the Cu complex significantly. This may be interpreted as the copper complex aiding the transfer of aspirin through an acid-base reaction at the LOX enzyme which subsequently blocks its binding.

Changes in fatty acid profile of Bovec sheep milk due to different pasture altitude.

Sheep rearing on mountain pastures is an ancestral tradition in northwestern Slovenia. The indigenous Bovec sheep are widespread there and are well adapted to the rough Alpine rearing conditions. Every year, after weaning, the sheep start grazing in the lowlands (L) and then gradually move to mountain pastures, and finally, to the highland (H) pastures of the Alps. Grazing positively affects the fatty acid (FA) composition in sheep milk fat with increased availability of polyunsaturated FA (PUFA) in grass, and subsequently, in milk. Consequently, the objective of this work was to study the FA profile in sheep milk during grazing in four geographical areas in the Alps. A total of 15 ewes of the Bovec sheep breed were randomly selected and milk samples from these ewes were taken at four different pasture locations that differed with regard to altitude: the L pasture location at an altitude of 480 m, the mountain pastures (M1 and M2) at altitudes of 1100 to 1300 m and 1600 to 1900 m, respectively, and the H pastures at altitudes of 2100 to 2200 m. Milk samples from the ewes were taken during the grazing season from April to September. The chemical and FA composition of the milk samples from each pasture location were determined. There were significant differences in the concentrations of FA among the L, M1, M2 and H milk samples. We observed decreases of the concentrations of saturated FA (SFA) in milk from L to H pastures. The concentration of α-linolenic FA, monounsaturated FA (MUFA), PUFA and n-3 PUFA in milk were increased significantly with pasture altitude. The n-6/n-3 PUFA ratio was reduced by the change of pasture altitude with the lowest value at the M1 pasture (1.5). The concentrations of total SFA decreased significantly and was lowest at the L pasture. Our results underline the importance of the effect of grazing in the Alpine region associated with pasture altitude on the FA profile of sheep milk. The first variation in FA concentration in sheep milk occurred between L and M1, although it was more evident on H pastures in the Alpine mountains. Changes of the FA profile in sheep milk due to pasture altitude were related to variation in FA concentration in the pasture and the botanical composition of the pasture location.

Strategies in the rational drug design.

Rational design is applied in the discovery of novel lead drugs. Its rapid development is mainly attributed to the tremendous advancements in the computer science, statistics, molecular biology, biophysics, biochemistry, medicinal chemistry, pharmacokinetics and pharmacodynamics experienced in the last few decades. The promising feature that characterizes the application of rational drug design is that it uses for developing potential leads in drug discovery all known theoretical and experimental knowledge of the system under study. The utilization of the knowledge of the molecular basis of the system ultimately aims to reduce human power cost, time saving and laboratory expenses in the drug discovery. In this review paper various strategies applied for systems which include: (i) absence of knowledge of the receptor active site; (ii) the knowledge of a homology model of a receptor, (iii) the knowledge of the experimentally determined (i.e. X-ray crystallography, NMR spectroscopy) coordinates of the active site of the protein in absence and (iv) the presence of the ligand will be analyzed.

Docking and Molecular Dynamics Calculations of Pyrrolidinone Analog MMK16 Bound to COX and LOX Enzymes.

The new molecule 4-[(2S)-2-(1H-imidazol-1-ylmethyl)-5-oxotetrahydro-1H-pyrrol-1-yl]methylbenzenecarboxylic acid (MMK16) was found to have promising anti-inflammatory activity. This biological behavior of MMK16 triggered our interest to study its binding affinity using NMR spectroscopy in LOX and its docking and molecular dynamics (MD) properties in LOX and COX enzymes. The present NMR and docking binding studies not only rationalize the obtained biological results since in all three receptors MMK16 shows high affinity and scoring but also make it a potential dual LOX-5/COX-2 inhibitor. Thus, this class of molecules must be further investigated for discovering compounds possessing better biological activity and more lasting biological effect.