DNA damage causes TP53-dependent coupling of self-renewal and senescence pathways in embryonal carcinoma cells

Comment on: Wu R, et al. Clin Cancer Res 2011; 17:7359–72     

Wolframin deficiency is accompanied with metabolic inflexibility in rat striated muscles

The protein wolframin is localized in the membrane of the endoplasmic reticulum (ER), influencing Ca2+ metabolism and ER interaction with mitochondria, but the exact role of the protein remains unclear. Mutations in Wfs1 gene cause autosomal recessive disorder Wolfram syndrome (WS). The first symptom of the WS is diabetes mellitus, so accurate diagnosis of the disease as WS is often delayed. In this study we aimed to characterize the role of the Wfs1 deficiency on bioenergetics of muscles. Alterations in the bioenergetic profiles of Wfs1-exon-5-knock-out (Wfs1KO) male rats in comparison with their wild-type male littermates were investigated using high-resolution respirometry, and enzyme activity measurements. The changes were followed in oxidative (cardiac and soleus) and glycolytic (rectus femoris and gastrocnemius) muscles. There were substrate-dependent alterations in the oxygen consumption rate in Wfs1KO rat muscles. In soleus muscle, decrease in respiration rate was significant in all the followed pathways. The relatively small alterations in muscle during development of WS, such as increased mitochondrial content and/or increase in the OxPhos-related enzymatic activity could be an adaptive response to changes in the metabolic environment. The significant decrease in the OxPhos capacity is substrate dependent indicating metabolic inflexibility when multiple substrates are available.     

Kinetic characteristics of interaction of conotoxin with N-type Ca2+ channels in rat hippocampal neurons

The binding and unbinding constants describing interaction of ω-CTx-GVIA with N-type Ca²⁺ channels were calculated based on the time course of the blocking action of the toxin. The experiments were carried out on pyramidal neurons freshly dissociated from theCA3 region of the rat hippocampus using a “concentration-clamp” technique and a patch-clamp technique in the whole-cell configuration. The bindingk ₁ and unbindingk ₋₁ constants were evaluated as 0.32 (μM·sec)⁻¹ and 0.004 sec⁻¹, respectively. The dissociation constantK _D kinetically derived from the ratiok ₋₁/k ₁ was 0.012 μM. These values allow us to interpret the apparent “irreversibility” of the toxin action.     

Morphological separation of aphid species <Emphasis Type="Italic">Cryptomyzus leonuri</Emphasis> Bozhko, 1961 and <Emphasis Type="Italic">Cryptomyzus alboapicalis</Emphasis> (Theobald, 1916) (Hemiptera: Sternorrhyncha: Aphididae)

Aphids of the species Cryptomyzus alboapicalis (Theobald, 1916) and Cryptomyzus leonuri Bozhko, 1961 are very similar morphologically, although exploit different host plants (Lamium album and Leonurus cardiaca, respectively). Morphological characters proposed for the separation of this species couple in the identification key to European Cryptomyzus species appeared to be of little discriminatory power when applied to apterous viviparous females from clonal lineages, whilst alate viviparous females of C. leonuri were not included in the key at all. The aim of this study was to find reliable morphological characters and their combinations for the separation of apterous and alate viviparous females of C. alboapicalis and C. leonuri. Forward stepwise discriminant analysis based on characters without statistically significant (P < 0.05) correlation (|r| ≥ 0.50) with body length resulted in canonical functions enabling correct classification of 95–100% of specimens from clonal lineages involved in the analysis with a priori specified group membership. The post hoc classification gave 95–100% correct identification of individuals from clonal and 85–100% from field-collected samples. The discriminative values of single morphological characters and canonical functions are discussed and modified key for the morphological identification of C. alboapicalis and C. leonuri apterous and alate viviparous females is suggested.     

Redox and metal ion binding properties of human insulin-like growth factor 1 determined by electrospray ionization mass spectrometry

Insulin-like growth factor 1 (IGF-1) is a 70-residue hormone containing three intramolecular disulfide bridges. IGF-1 and other growth factors are oxidatively folded in the endoplasmic reticulum and act primarily in the blood, under relatively oxidative conditions. It is known that IGF-1 exists in various intracellular and extracellular compartments in the oxidized form; however, the reduction potential of IGF-1 and the ability of fully reduced IGF-1, which contains six cysteine residues, to bind transition metal ions are not known. In this work, we determine that the redox potential of human IGF-1 is equal to -332 mV and the reduced form of hIGF-1 can bind cooperatively four Cu(+) ions, most probably into a tetracopper-hexathiolate cluster. The Cu(+) binding affinity of hIGF-1 is, however, approximately 3 times lower than that for the copper chaperones; thus, we can conclude that fully reduced hIGF-1 cannot compete with known Cu(+)-binding proteins.     

Pharmacological properties of the potassium-activated inward current in pyramidal hippocampal neurons

Elevation of the external potassium concentration induced a two-phase inward current in freshly isolated pyramidal hippocampal neurons. This current was voltage-dependent and demonstrated strong inward rectification. The current consisted of a leakage current and a time-dependent current (τ=40–50 msec at 21°C); the latter was designated asI _ΔK. As was shown earlier, K⁺ is a major charge carrier in the development of slow potassium-activated current. The pharmacological properties ofI _ΔK were studied using a patch-clamp technique.I _ΔK was completely blocked by external 10 mM TEA or 5 mM Ba²⁺ (IC₅₀=480±90mM) and exhibited low sensitivity to extracellular Cs⁺ (2 mM). This current was not affected by 1 mM 4-aminopyridine and was insensitive to a muscarinic agonist, carbachol (50 μM), and to 1 mM extracellular Cd²⁺. Elevation of external Ca²⁺ from 2.5 mM to 10 mM did not changeI _ΔK. Our data indicate that the pharmacological properties ofI _ΔK differ from those of other voltage-gated potassium currents, but more specific blockers must be used to make this evidence conclusive.