The binding of ethyl isocyanide to ferroperoxidase

The equilibrium and kinetics of ethyl isocyanide binding to ferroperoxidase were studied. At pH9.1 the results of both studies are consistent with a single-process model with an affinity constant of 95m(-1) and combination and dissociation constants of 2.2x10(3)m(-1).s(-1) and 23s(-1) respectively. Ethyl isocyanide is not bound significantly at pH values lower than 6.0, and in this behaviour and the pH-dependence of the affinity constant, similarities exist between isocyanide and cyanide binding. The enthalpy of the process measured by equilibrium methods is -59kJ/mol (-14kcal/mol). At pH values below 9, the ethyl isocyanide adduct changes in a slow time-dependent manner, giving rise to a new species. These changes are reversible on increasing the pH. The results are discussed in relation to other known information about ligand binding to ferroperoxidase and to myoglobin.     

Permanent parahisian pacing

Right Ventricular Apical permanent pacing could have negative hemodynamic effects. A physiologic pacing modality should preserve a correct atrio-ventricular and interventricular synchronization. This can be obtained through biventricular pacing, left ventricular pacing, or from alternative right ventricular pacing sites. Direct His Bundle Pacing (DHBP) was documented as reliable and effective for preventing the desynchronization and negative effects of right ventricular apical pacing. It is, however, a complex method that requires longer average implant times, cannot be carried out on all patients and presents high pacing thresholds. On the contrary, the parahisian pacing, with simpler feasibility and reliability criteria, seems to guarantee an early invasion of the His-Purkinje conduction system, with a physiological ventricular activation, very similar to the one that can be obtained with direct His bundle pacing. We present our experience on 68 patients who underwent a permanent right ventricular pacing in hisian/parahisian region, for advanced AV block and narrow QRS. In the first 17 patients we performed a double-blind randomized controlled study, with two 6-months cross-over periods in parahisian and apical pacing, documenting a significant improvement of NYHA class, exercise tolerance, quality of life score, mitral and tricuspidal regurgitation degree, and interventricular mechanical delay. In the subsequent 51 patients, in a mean follow of 21 months/patient, the pacing threshold remained stable (0.7+/-0.5 V implant; 0.9+/-0.7 V follow-up; p=0.08). The ejection fraction maintained medium-long term stable values, confirming the fact that the parahisian pacing can prevent deterioration of the left ventricular function. Parahisian pacing, therefore, has proven to be a reliable method, easy to apply and effective in preventing the negative effects induced by non-physiological right ventricular apical pacing.     

The direct permeation of nanoparticles through the plasma membrane transiently modifies its properties

The exposure to metal nanoparticles (NPs) has increased with their widespread use in industry, research and medicine. It is well known that NPs may enter cells and that this mechanism is crucial to exert both the therapeutic and toxicity effects. The main cellular entrance route is endocytosis-based, however, recent experimental studies, have reported that NPs can also enter the cell crossing directly the plasma membrane, it is thus important to investigate this alternative internalization mechanism. Size, surface chemistry, solubility and shape play a role in NP ability of entering the cell, but it is still to be elucidated how these properties act on cell membrane. We have demonstrated that a direct permeation of metal oxide NPs through the lipid bilayer of the cell membrane can occur, giving direct access to the cytoplasm. In this paper, using the powerful tool of Xenopus laevis oocytes and two electrode Voltage Clamp, we have investigated several parameters that can influence the direct crossing. The most significant of them is the NP hydrodynamic size as clearly shown by the comparison of the behaviour between Co₃O₄ and NiO NPs. By collecting biophysical membrane parameters in different conditions, we have shown that NPs that are able to cross the membrane share the ability to maintain a hydrodynamic size lower than 200 nm. The presence of this route of entrance must be considered for a better comprehension of the effect at intracellular level considering possible mechanism in order to a safer design of engineered NPs.     

Isoelectric focusing of human head hair keratins from various racial groups

Isoelectric focusing of extracted keratin not S-carboxymethylated followed by silver staining was used in this study with the attempt to identify head hair keratins from different racial groups: Guatuso and Caribbean (Costa Rica, America), Balanta (Guinea-Bissau, Africa) and Sardinian (Italy, Europe). Morphological analysis of hair and quantitation of solubilized total proteins were also performed. Keratins extracted from hair gave similar IEF patterns for all samples in the ranges of pH 4.5–5.0 and 5.4–7.4, differing only in the intensity and width of the bands. IEF patterns for Guatuso samples differ from all the others in the presence of some additional bands in the range of pH 4.0–4.5. Different electrophoretic patterns are not associated with detectable differences in the morphology of the fibers. It seems to us that electrophoretic techniques may be useful in the identification of hair from different racial groups.     

Absence of 2009 Pandemic H1N1 Influenza A Virus in Fresh Pork

The emergence of the pandemic 2009 H1N1 influenza A virus in humans and subsequent discovery that it was of swine influenza virus lineages raised concern over the safety of pork. Pigs experimentally infected with pandemic 2009 H1N1 influenza A virus developed respiratory disease; however, there was no evidence for systemic disease to suggest that pork from pigs infected with H1N1 influenza would contain infectious virus. These findings support the WHO recommendation that pork harvested from pandemic influenza A H1N1 infected swine is safe to consume when following standard meat hygiene practices.     

Denaturation of Aplysia myoglobin. Equilibrium study

The effect of temperature and solvent composition on the reversible denaturation of Aplysia myoglobin is reported here. The denaturation is accompanied by changes in the haem absorption spectrum, in the intrinsic fluorescence of the protein and in its optical activity in the Soret and ultraviolet regions. Under all conditions examined (i.e. different temperatures, different concentrations of various alcohols and different concentrations of proteins) the system appears to be in a state of true equilibrium and good agreement is found between thermally-induced and solvent-induced transitions.     

Uptake of lead,cadmium and zinc by the fairy shrimp,Branchinecta longiantenna (Crustacea: Anostraca)

Individuals of the fairy shrimp, Branchinecta longiantenna, were subjected to 5 concentrations (0.1 to 15 mg l^–1) of Pb in diluted habitat water at 13 °C. Lead concentrations (mg kg^–1 wet weight) in the animals were determined at 2-day intervals by digestion in nitric acid followed by atomic absorption analysis. The shrimp were also subjected to 0.1 mg l^–1 media of Cd and Zn, separately.Uptake rates by the fairy shrimp for the three metal ions at 0.1 mg l^–1 were: 0.111, 0.0885, and 0.0460 mg kg^–1 day^–1 for Zn, Pb, and Cd, respectively. After 2 days in 1.0 mg l^–1 Cd or Zn, the animals expired; but they surviced for 8 days in a 10 mg l^–1 Pb medium and for 2 days in 25 mg l^–1 Pb. Lead uptake demonstrated a linear dependence on the Pb concentration in the media.Shrimp survived at much higher tissue accumulations of Pb compared to Zn and Cd. Estimated lethal doses were 20, 1.2–2.4, and 0.4–1.4 mg kg^–1 wet weight for Pb, Zn, and Cd, respectively. Pb was found to be at much lower concentration than Cd or Zn in the natural pond water but between Cd and Zn levels in the sediment. Thus Cd and Zn probably present a greater threat to B. longiantenna than Pb, although Pb may be in higher concentration in the environment.Contribution 47, Laboratory of Ecology, The Claremont Colleges, Claremont, CA 91711, USA. Send reprint requests to Clyde Eriksen.     

Guanosine modulates SUMO2/3-ylation in neurons and astrocytes via adenosine receptors

Purinergic Signalling - SUMOylation is a post-translational modification (PTM) whereby members of the Small Ubiquitin-like MOdifier (SUMO) family of proteins are conjugated to lysine residues in...