Metabolic syndrome in an island population of the eastern Adriatic coast of Croatia

The metabolic syndrome, an assemblage of risk factors, viz., obesity, dyslipidemia, carbohydrate intolerance, and hypertension, associated with development of cardiovascular diseases and Type 2 diabetes, has become a major public health problem in the developed countries. However, data on its prevalence in worldwide populations, particularly in non-industrialized societies is sparse. We report the prevalence of metabolic syndrome in an island population of the eastern Adriatic coast of Croatia, a distinctly homogenous population living in relative isolation with a traditional way of life style pattern. The participants consist of 381 unrelated individuals (136 males, 245 females) from the island of Hvar, age 19 and above. Hvar is among the most populated Croatian islands with a total population of 11,459 individuals according to the 1991 census. Metabolic syndrome was assessed following the National Cholesterol Education Program (NCEP) criteria, with the exception of using body mass index and waist hip ratio as the predictors of obesity. Overall age-adjusted prevalence of metabolic syndrome is 26% (32% in males and 24% in females) with body mass index, and 42% (57% in males and 36% in females) with waist hip ratio as the measures of obesity. Pairwise correlations of the age and sex-adjusted individual components show that with the exception of fasting plasma glucose, the other components of metabolic syndrome are significantly associated with each other, suggesting their co-morbidity. In spite of adherence to a relatively traditional life-style pattern together with a "Mediterranean diet" and rural habitat, the prevalence of metabolic syndrome is substantially high in the population of Hvar. It is likely that factors other than nutritional practices, that might include genetic susceptibility, could potentially be important in predisposition to metabolic syndrome.     

Seasonal dynamics of foliar antioxidative enzymes and total anthocyanins in natural populations of Iris pumila L.

Aims Plants in their natural habitats frequently cope with a multitude of abiotic stresses, such as high light intensity, extreme temperatures and water deficit, which often co-occur during periods of drought, especially in semi-arid and arid regions. Exposure of plants to stressful environmental conditions usually induce overproduction of reactive oxygen species (ROS) that, as highly toxic derivatives of O 2, can assault all cell macromolecules, leading to the disruption of cellular homeostasis and, consequently, the uncoupling of major metabolic processes, the photosynthesis and photorespiration. In order to minimize ROS-mediated cellular damage, plants have evolved highly efficient antioxidative defense systems that include both enzymatic and non-enzymatic components. Since abiotic stress can also operate as a strong evolutionary force that shapes adaptations in natural plant populations, the aim of this study was to examine the seasonal variation patterns of two distinct antioxidative systems, ROS-scavenging enzymes and anthocyanin pigments, in the leaf tissue of a steppe plant, Iris pumila, as expressed under contrasting light conditions that the species regularly experiences in the wild.Methods We selected two natural populations of I.pumila inhabiting the alternative radiation environments in the Deliblato Sands, a sun-exposed dune site and a woodland understory. The specific activity of three antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) and the content of total anthocyanins were examined in leaves of I.pumila plants collected from each of the 31 Iris clones (17 in the exposed population and 14 in the shaded population) once during each of the three seasons, spring, summer and autumn in 2004. Specifically, a fully expanded leaf was cut from each clonal plant between 15:00 and 16:00 h, immediately frozen in liquid nitrogen and stored at ?70°C until preparation.Important findings Generally, all three antioxidative enzymes were up-regulated in summer-harvested leaves compared to their spring or autumn counterparts, as was observed for the concentration of foliar anthocyanins, indicating that strengthening of antioxidant systems was the key mechanism for long-term acclimatization of I.pumila plants to stressful environmental conditions within their natural ecological niches. When plants from contrasting radiation environments were compared, SOD and CAT activities appeared to be greater in shade-exposed than in sun-exposed leaves. Conversely, POD activity and the content of foliar anthocyanins were notably higher in foliage experiencing full sunlight relative to those developed under vegetation canopy, suggesting the synergistic function of these two molecules in protecting leaf cells against photoinhibitory and photooxidative effects of strong light.     

(19)F NMR studies of the leucine-isoleucine-valine binding protein: evidence that a closed conformation exists in solution

The leucine-isoleucine-valine binding protein (LIV) found in the periplasmic space of E. coli has been used as a structural model for a number of neuronal receptors. This "venus fly trap" type protein has been characterized by crystallography in only the open form. Herein we have labeled LIV with 5-fluorotryptophan (5F-Trp) and difluoromethionine (DFM) in order to explore the structural dynamics of this protein and the application of DFM as a potential (19)F NMR structural probe for this family of proteins. Based on mass spectrometric analysis of the protein overproduced in the presence of DFM, approximately 30% of the five LIV methionine residues were randomly substituted with the fluorinated analog. Urea denaturation experiments imply a slight decrease in protein stability when DFM is incorporated into LIV. However, the fluorinated methionine did not alter leucine-binding activity upon its incorporation into the protein. Binding of L-leucine stabilizes both the unlabeled and DFM-labeled LIV, and induces the protein to adopt a three-state unfolding model in place of the two-state process observed for the free protein. The (19)F NMR spectrum of DFM-labeled LIV gave distinct resonances for the five Met residues found in LIV. 5F-Trp labeled LIV gave a well resolved spectrum for the three Trp residues. Trp to Phe mutants defined the resonances in the spectrum. The distinct narrowing in line width of the resonances when ligand was added identified the closed form of the protein.     

Linking oxidative and salinity stress tolerance in barley: can root antioxidant enzyme activity be used as a measure of stress tolerance?

Aims

A causal relationship between salinity and oxidative stress tolerance and a suitability of using root antioxidant activity as a biochemical marker for salinity tolerance in barley was investigated.

Methods

Net ion fluxes were measured from the mature zone of excised roots of two barley varieties contrasting in their salinity tolerance using non-invasive MIFE technique in response to acute and prolonged salinity treatment. These changes were correlated with activity of major antioxidant enzymes; ascorbate peroxidase, catalase, and superoxide dismutase.

Results

It was found that genotypic difference in salinity tolerance was largely independent of root integrity, and observed not only for short-term but also long-term NaCl exposures. Higher K⁺ retention ability (and, hence, salinity tolerance) positively correlated with oxidative stress tolerance. At the same time, antioxidant activities were constitutively higher in a sensitive but not tolerant variety, and no correlation was found between SOD activity and salinity tolerance index during large-scale screening.

Conclusion

Although salinity tolerance in barley correlates with its oxidative stress tolerance, higher antioxidant activity at one particular time does not correlate with salinity tolerance and, as such, cannot be used as a biochemical marker in barley screening programs.     

Mechanism for Active Membrane Fusion Triggering by Morbillivirus Attachment Protein

The paramyxovirus entry machinery consists of two glycoproteins that tightly cooperate to achieve membrane fusion for cell entry: the tetrameric attachment protein (HN, H, or G, depending on the paramyxovirus genus) and the trimeric fusion protein (F). Here, we explore whether receptor-induced conformational changes within morbillivirus H proteins promote membrane fusion by a mechanism requiring the active destabilization of prefusion F or by the dissociation of prefusion F from intracellularly preformed glycoprotein complexes. To properly probe F conformations, we identified anti-F monoclonal antibodies (MAbs) that recognize conformation-dependent epitopes. Through heat treatment as a surrogate for H-mediated F triggering, we demonstrate with these MAbs that the morbillivirus F trimer contains a sufficiently high inherent activation energy barrier to maintain the metastable prefusion state even in the absence of H. This notion was further validated by exploring the conformational states of destabilized F mutants and stabilized soluble F variants combined with the use of a membrane fusion inhibitor (3g). Taken together, our findings reveal that the morbillivirus H protein must lower the activation energy barrier of metastable prefusion F for fusion triggering.