Yeast holoenzyme of protein kinase CK2 requires both β and β′ regulatory subunits for its activity

Protein kinase CK2 is a highly conserved Ser/Thr protein kinase that is ubiquitous among eucaryotic organisms and appears to play an important role in many cellular functions. This enzyme in yeast has a tetrameric structure composed of two catalytic (α and/or α′) subunits and two regulatory β and β′ subunits. Previously, we have reported isolation from yeast cells four active forms of CK2, composed of αα′ββ′, α₂ββ′, α′₂ββ′ and a free α′-catalytic subunit. Now, we report that in Saccharomyces cerevisiae CK2 holoenzyme regulatory β subunit cannot substitute other β′ subunit and only both of them can form fully active enzymatic unit. We have examined the subunit composition of tetrameric complexes of yeast CK2 by transformation of yeast strains containing single deletion of the β or β′ regulatory subunits with vectors carrying lacking CKB1 or CKB2 genes. CK2 holoenzyme activity was restored only in cases when both of them were present in the cell. Additional, co-immunoprecypitation experiments show that polyadenylation factor Fip1 interacts with catalytic α subunits of CK2 and interaction with beta subunits in the holoenzyme decreases CK2 activity towards this protein substrate. These data may help to elucidate the role of yeast protein kinase CK2β/β′ subunits in the regulation of holoenzyme assembly and phosphotransferase activity.     

Two new end-to-end single dicyanamide bridged Cu(II) complexes with Schiff base ligands: Structural, electrochemical and magnetic properties

The synthesis, crystal structures and magnetic properties of two different copper(II) complexes of formula [Cu(L¹)(dca)]_n · nClO₄ (1) and [Cu(L²)]₂(dca)(ClO₄) (2) [L¹ = N,N-dimethylethylene-N′-(pyridine-2-carbaldiiminato), HL² = N,N-dimethylethylene-N′-salicylaldiiminato, dca = dicyanamide anion] are described. Spectroscopic and electrochemical properties have also been discussed. A one-dimensional chain structure with single, symmetrical, μ_1,5-dca bridges is found in compound 1. The copper atom in 1 has a square pyramidal geometry. A tridentate Schiff base ligand, having NNN donor sites, and one nitrogen atom from dca occupy the basal plane. N(18) of a neighbouring unit occupies the apical site. The Schiff base used in compound 2 is a tridentate anion with NNO donor sites, which changes the structure in a dinuclear unit of copper atoms bridged by single end-to-end dicyanamide ion. The environment around copper in 2 is square planar. Magnetic susceptibility measurements for 1 and 2 reveal the occurrence of weak antiferromagnetic interaction through the dca ligand.     

Validated spectrofluorimetric and spectrophotometric methods for the determination of brimonidine tartrate in ophthalmic solutions via derivatization with NBD‐Cl. Application to stability study

Two simple, selective and accurate methods were developed and validated for the determination of brimonidine tartrate (BT) in pure state and pharmaceutical formulations. Both methods are based on the coupling of the drug with 4‐chloro‐7‐nitro‐2,1,3‐benzoxadiazole in borate buffer (pH 8.5) at 70 °C and measurement of the reaction product spectrophotometrically at 407 nm (method I) or spectrofluorimetrically at 528 nm upon excitation at 460 nm (method II). The calibration graphs were rectilinear over the concentration ranges of 1.0–16.0 and 0.1–4.0 µg/mL with lower detection limits of 0.21 and 0.03, and lower quantification limits of 0.65 and 0.09 µg/mL for methods I and II, respectively. Both methods were successfully applied to the analysis of commercial ophthalmic solution with mean recovery of 99.50 ± 1.00 and 100.13 ± 0.71%, respectively. Statistical analysis of the results obtained by the proposed methods revealed good agreement with those obtained using a comparison method. The proposed spectrofluorimetric method was extended to a stability study of BT under different ICH‐outlined conditions such as alkaline, acidic, oxidative and photolytic degradation. Furthermore, the kinetics of oxidative degradation of the drug was investigated and the apparent first‐order reaction rate constants, half‐life times and Arrhenius equation were estimated. The proposed methods are practical and valuable for routine applications in quality control laboratories for the analysis of BT. Copyright © 2014 John Wiley & Sons, Ltd.     

Prehibernation and hibernation effects on the D-3-hydroxybutyrate dehydrogenase of the heavy and light mitochondria from liver jerboa (Jaculus orientalis) and related metabolism

The D-3-hydroxybutyrate dehydrogenase (BDH) (EC 1.1.1.30) from liver jerboa (Jaculus orientalis), a ketone body converting enzyme in mitochondria, in two populations of mitochondria (heavy and light) has been studied in different jerboa states (euthermic, prehibernating and hibernating). The results reveal: (1) important variations between states in terms of ketones bodies, glucose and lipid levels; (2) significant differences between the BDH of the two mitochondrial populations in term of protein expression and kinetic properties. These results suggest that BDH leads an important conformational change depending on the physiological state of jerboa. This BDH structural change could be the consequence of the lipid composition modifications in inner mitochondrial membrane leading to changes in BDH catalytic properties.     

Influence of age on the bioaccumulation of heavy metals in Apodemus sylvaticus at Merja Zerga lagoon,Morocco

The influence of age and sex on the bioaccumulation of heavy metals in Apodemus sylvaticus was studied in Merja Zerga lagoon in northern Morocco. Five trace metal elements (Zn, Pb, Cr, Cu and Fe) were quantitatively analyzed by Varian AA 240 atomic absorption spectroscopy with graphite furnace in three organs (Liver, Kidney and Heart) from animals of different age and sex. The maximum metal level of the analyzed samples was recorded in adults and was limited to 46.62 μg/g for Pb and 35.1 μg/g for Cu, while it reached 22.69 μg/g, 7.59 μg/g and 6.78 μg/g for Cr, Zn and Fe, respectively. Highly significant differences were found for bioaccumulation of heavy metals according to animal ages and no significant differences were observed between the two sexes among the studied animals. Our results revealed also the existence of a strong correlation (r > 0.65) between the majority of biometric parameters and the trace element concentrations. In general, we found that age is a critical factor in estimating the level of heavy metal pollution. Other characteristics such as habitat, feeding habits and anti-predator behavior of the species need to be studied.     

Estimation of aboveground biomass in conserved areas of Stipa tenacissima L. stands in the high steppes of western Algeria by mean of the Landsat 8 imagery‐based vegetation indices

The main aim of this paper was to evaluate the use of OLI spectral data as a tool to assess the steppe vegetation in a conservation context. The field sampling was conducted for two specific areas of treatment (a) an exclosure area and (b) a free grazing area. After testing several vegetation indices (VIs), the optimal results were obtained for the Normalised Difference Vegetation Index (NDVI)‐based aboveground biomass model with r² = 0.61 and r² = 0.72 for total and perennial biomass, respectively. No difference between observed and predicted total and perennial biomass was found (p = 0.700 and p = 0.306, respectively). The comparison between the two treatments using the field sampling revealed a significant difference on total plant cover (p = 0.016) and total biomass (p = 0.005), with a plant cover of 50.6% and a biomass of 325.771 kg dry matter per hectare (kg DM ha^?1) on average in grazed area and 66.9%, 1,407.869 kg DM ha^?1 in exclosure. Finally, a concordance is noted between the results obtained by the NDVI‐based biomass model and the field sampling‐based biomass.     

The influence of postnatal nutrition on lipoprotein lipase activity and hormone sensitive lipolysis in vitro of rat adipose tissue

In rats grown up in small and large nests the lipolytic activity of adipose tissue was studied in vitro in dependence on age. Independent of age and sex a significantly higher lipoprotein lipase activity as well as a lower basal or norepinephrine stimulated lipolysis and reesterification (absolute) occurred in rats from small nests compared with rats from large nests. The calculated degree of reesterification was independent of the nest size, but decreased from 80% to 30% from one month to one year of age. The marked differences in the lipolytic activities of adipose tissue emphasize the distinct influence of the post-natal nutrition on metabolic functions in the later life and lead to the conclusion that the metabolism of adipose tissue of animals from small nests is directed towards a long-term increased storage of lipids.