Structure-functional organization of eukaryotic high-affinity copper importer CTR1 determines its ability to transport copper, silver and cisplatin

It was shown recently, that high affinity Cu(I) importer eukaryotic protein CTR1 can also transport in vitro abiogenic Ag(I) ions and anticancer drug cisplatin. At present there is no rational explanation how CTR1 can transfer platinum group, which is different by coordination properties from highly similar Cu(I) and Ag(I). To understand this phenomenon we analyzed 25 sequences of chordate CTR1 proteins, and found out conserved patterns of organization of N-terminal extracellular part of CTR1 which correspond to initial metal binding. Extracellular copper-binding motifs were qualified by their coordination properties. It was shown that relative position of Met- and His-rich copper-binding motifs in CTR1 predisposes the extracellular CTR1 part to binding of copper, silver and cisplatin. Relation between tissue-specific expression of CTR1 gene, steady-state copper concentration, and silver and platinum accumulation in organs of mice in vivo was analyzed. Significant positive but incomplete correlation exists between these variables. Basing on structural and functional peculiarities of N-terminal part of CTR1 a hypothesis of coupled transport of copper and cisplatin has been suggested, which avoids the disagreement between CTR1-mediated cisplatin transport in vitro, and irreversible binding of platinum to Met-rich peptides.     

Cholesterol reduces membrane electroporation and electric deformation of small bilayer vesicles

Electric fields, similar in the order of magnitude of the natural membrane fields of cellular lipid/protein membranes, and chemical relaxation spectrometry can be used as tools to quantify the rigidifying effect of cholesterol in membranes. Small unilamellar vesicles of radius a=50+/-3 nm, prepared form phosphatidylcholine, phosphatidylserine and phosphatidyl-glycerol in the molar ratio 1:1:1 and containing the optical lipid probe molecule 2-(3-diphenyl-hexatrienyl) propanoyl)-1-palmitoyl-sn-glycerol-3-phosphocholine (beta-DPH pPC), serve as examples for curved lipid membranes. The data of electrooptical turbidity and absorbance relaxations at the wavelength lambda=365 nm are analysed in terms of membrane bending rigidity kappa and membrane stretching modulus K. Both kappa and K increase with increasing mole fraction x of cholesterol up to x=0.5. The cholesterol induced denser packing of the lipids reduces the extent of both membrane electroporation (ME) and electroelongation of the vesicles. Further on, cholesterol in the lipid phase and sucrose in the aqueous suspension reduce the extent of membrane undulation and electro-stretching.     

Systematics on the threshold of the XXI century. Traditional principles and fundamentals from today's point of view

Systematics as regarded is a purely theoretical domain of biology, and its product, system, as a specific biological theory, or a topologo-genetic model of the biota. Linnaeus was the first to introduce the idea of system and the systematic approach into the natural history. The advent of evolutionism brought new meaning to the old term "affinity", so Linnaeus' slogan of natural system got new life, and Linnaeus taxonomy assimilated the evolutionary ideology quite naturally and much easier than many other departments of biology. The difference between natural and artificial systems is remaining, and it is in their goals, as formulated by Linnaeus: heuristic of the former and cataloguing of the latter. Linnaeus' clairvoyance discovered the existence of an infrageneric level of genetic integration provable by naturalists' experience. He chose for it the designation of "species" and laid it down as primary, basic unit of his system. This is plainly evident from his own writings; the story about Linnaean species being products of a logical division of genera is a pure fiction. Modern populational model of species, by 3 important criteria, appears to be more akin to the Linneaean one than to the ideas of Lamarckism and early Darwinism. Systematic approach focuses rather on the interrelations among elements and their relative position, then on the properties and qualities of separately treated individual elements. In the development of systematics the aspect of "nexification" (study of connections) has been continuously gaining attention especially regarding the nomenclature where connotation has been totally forced out by denotation.     

What do over-trained athletes and patients with neurodegenerative diseases have in common? Mitochondrial dysfunction

Under pathological conditions and excessive stress, mitochondria may experience a severe and irreversible loss of function. Both strenuous exhaustive exercise and neurodegenerative disorders appear to share defects in mitochondrial function that may fiercely disrupt the integrity and homeostasis of the organelle, leading to perennial pathological substrates. Here, we overview similarities of mitochondrial dysfunction in two conditions and discuss possible areas of interdisciplinary collaboration and research translation between sports medicine and neurology.     

Microsatellite Variation in Populations of Atlantic Salmon from North Europe

Our aim was to investigate the level of genetic differentiation in northern European populations of Atlantic salmon, to establish the genetic relationship among major salmon populations in Russia and North Norway, and to compare these to populations from the western Atlantic lineage. Samples were collected along an east—west axis, from Pechora River in Russia to Restigouche River in Quebec, Canada. A total of 439 individual salmon were collected from seven rivers (sample sizes from 50 to 84 individuals). The samples were analysed for variation at four microsatellite loci; Ssa13.37, Ssa14, Ssa171 and Ssa171. Significant differences were found between most of the European populations, and the populations from the Tana and Pechora Rivers were most distinct. The samples from the Rivers Mezenskaya Pizhma and Emtsa in Arkhangelsk oblast in Russia were not significantly different from each other in an exact test of population differences. All other river pairs were significantly different. These results confirmed the deep genetic divergence between American and European salmon populations demonstrated in earlier studies, with alleles specific to continent found in three of the microsatellites.     

Guanine-derived radicals: dielectric constant-dependent stability and UV/Vis spectral properties: a DFT study

Upon successive deprotonation of the guanine radical cation, various neutral radicals and radical anions can be formed. Their relative stability and UV/Vis absorption spectra have been calculated by DFT in the vacuum and in aqueous solution. Good agreement with experimental data is obtained when solvent effects are taken into account. The experimental observation that in the nucleosides deprotonation of the guanine radical cation occurs at N1 (formation of N1G(*)) in water and at N2 (formation of N2G(*)) in single crystals is now explained by a strong effect of the dielectric constant of the environment on their stability. While SCRF=PCM and CPCM (Gaussian 03) describe the trend, SCRF=DPCM (Gaussian 98) even shows the crossover from N2G(*) to N1G(*) at high dielectric constant. A crossover of the preferred deprotonation site is also given by the nucleoside itself. While for the gas phase a deprotonation at N2 is calculated to be favored over that at N1, the reverse is found for an aqueous environment (in agreement with the experiment). The radical anions of guanine, N9N1G(*)(-) and N9N2G(*)(-), are very similar in energy, but a comparison of the experimental and calculated UV/Vis spectra allows us to identify the experimentally observed intermediate clearly as N9N1G(*)(-).     

Serum but not myocardial TNF-alpha concentration is increased in pacing-induced heart failure in rabbits

In animals and patients with severe heart failure (HF), the serum tumor necrosis factor-alpha (TNF-alpha) concentration is increased. It is, however, still controversial whether or not such increased serum TNF-alpha originates from the heart itself or is of peripheral origin secondary to gastrointestinal congestion and increased endotoxin concentration. We therefore now examined TNF-alpha in serum, myocardium, and liver of sham-operated and HF rabbits. In nine rabbits in which HF was induced by left ventricular (LV) pacing at 400 beats/min for 3 wk, LV end-diastolic diameter was increased and systolic shortening fraction (9.4 +/- 1.0 vs. 28.5 +/- 1.3%, echocardiography, P < 0.05) was reduced. Serum TNF-alpha was higher in HF than in sham-operated rabbits (240 +/- 24 vs. 150 +/- 22 U/ml, WEHI-cell assay, P < 0.05). In the heart, TNF-alpha was located mainly in the vascular endothelium (immunohistochemistry), and TNF-alpha protein (920 +/- 160 vs. 900 +/- 95 U/g) did not differ between groups. In the liver of HF rabbits, hepatocytes expressed TNF-alpha, and TNF-alpha protein was increased compared with sham-operated rabbits (2,390 +/- 310 vs. 1,220 +/- 135 U/g, P < 0.05) and correlated to the number of hepatic leukocytes (r = 0.85) and serum TNF-alpha (r = 0.69). The intestinal endotoxin concentration was 24.5 +/- 1.2 vs. 17.0 +/- 3.1 endotoxin units/g wet wt (P < 0.05) in HF compared with sham-operated rabbits. In this HF model, serum but not myocardial TNF-alpha is increased. The increased serum TNF-alpha originates from peripheral sources.     

CD2AP/CIN85 balance determines receptor tyrosine kinase signaling response in podocytes

Defects in podocyte signaling are the basis of many inherited glomerular diseases leading to glomerulosclerosis. CD2-associated protein (CD2AP) is highly expressed in podocytes and is considered to play an important role in the maintenance of the glomerular slit diaphragm. Mice deficient for CD2AP (CD2AP(-/-)) appear normal at birth but develop a rapid onset nephrotic syndrome at 3 weeks of age. We demonstrate that impaired intracellular signaling with subsequent podocyte damage is the reason for this delayed podocyte injury in CD2AP(-/-) mice. We document that CD2AP deficiency in podocytes leads to diminished signal initiation and termination of signaling pathways mediated by receptor tyrosine kinases (RTKs). In addition, we demonstrate that CIN85, a paralog of CD2AP, is involved in termination of RTK signaling in podocytes. CIN85 protein expression is increased in CD2AP(-/-) podocytes in vitro. Stimulation of CD2AP(-/-) podocytes with various growth factors, including insulin-like growth factor 1, vascular endothelial growth factor, and fibroblast growth factor, resulted in a significantly decreased phosphatidylinositol 3-kinase/AKT and ERK signaling response. Moreover, increased CIN85 protein is detectable in podocytes in diseased CD2AP(-/-) mice, leading to decreased base-line activation of ERK and decreased phosphorylation after growth factor stimulation in vivo. Because repression of CIN85 protein leads to a restored RTK signaling response, our results support an important role of CD2AP/CIN85 protein balance in the normal signaling response of podocytes.     

Two primate-specific small non-protein-coding RNAs in transgenic mice: neuronal expression, subcellular localization and binding partners

In a rare occasion a single chromosomal locus was targeted twice by independent Alu-related retroposon insertions, and in both cases supported neuronal expression of the respective inserted genes encoding small non-protein coding RNAs (npcRNAs): BC200 RNA in anthropoid primates and G22 RNA in the Lorisoidea branch of prosimians. To avoid primate experimentation, we generated transgenic mice to study neuronal expression and protein binding partners for BC200 and G22 npcRNAs. The BC200 gene, with sufficient upstream flanking sequences, is expressed in transgenic mouse brain areas comparable to those in human brain, and G22 gene, with upstream flanks, has a similar expression pattern. However, when all upstream regions of the G22 gene were removed, expression was completely abolished, despite the presence of intact internal RNA polymerase III promoter elements. Transgenic BC200 RNA is transported into neuronal dendrites as it is in human brain. G22 RNA, almost twice as large as BC200 RNA, has a similar subcellular localization. Both transgenically expressed npcRNAs formed RNP complexes with poly(A) binding protein and the heterodimer SRP9/14, as does BC200 RNA in human. These observations strongly support the possibility that the independently exapted npcRNAs have similar functions, perhaps in translational regulation of dendritic protein biosynthesis in neurons of the respective primates.