Evidence that prokineticin receptor 2 exists as a dimer in vivo

Prokineticins are proteins that regulate diverse biological processes including gastrointestinal motility, angiogenesis, circadian rhythm, and innate immune response. Prokineticins bind two closed related G-protein coupled receptors (GPCRs), PKR1 and PKR2. In general, these receptors act as molecular switches to relay activation to heterotrimeric G-proteins and a growing body of evidence points to the fact that GPCRs exist as homo- or heterodimers. We show here by Western-blot analysis that PKR2 has a dimeric structure in neutrophils. By heterologous expression of PKR2 in Saccharomyces cerevisiae, we examined the mechanisms of intermolecular interaction of PKR2 dimerization. The potential involvement of three types of mechanisms was investigated: coiled-coil, disulfide bridges, and hydrophobic interactions between transmembrane domains. Characterization of differently deleted or site-directed PKR2 mutants suggests that dimerization proceeds through interactions between transmembrane domains. We demonstrate that co-expressing binding-deficient and signaling-deficient forms of PKR2 can re-establish receptor functionality, possibly through a domain-swapping mechanism.     

Investigations of a nanostructured FeMnSi shape memory alloy produced via severe plastic deformation

Low-cost iron-based shape memory alloys (SMAs) show great potential for engineering applications. The developments of new processing techniques have recently enabled the production of nanocrystalline materials with improved properties. These developments have opened avenues for newer applications for SMAs. The influence of severe plastic deformation induced by the high-speed high-pressure torsion (HSHPT) process on the microstructural evolution of an Fe–Mn–Si–Cr alloy was investigated. Transmission electron microscopic analysis of the alloy revealed the existence of nanoscale grains with an abundance of stacking faults. The high density of dislocations characteristic of severe plastic deformation was not observed in this alloy. X-ray diffraction studies revealed the presence of ε-martensite with an HCP crystal structure and γ-phase with an FCC structure.     

Hybrid formation in mixtures of globins from different species

Riassunto Miscele di globine preparate da emoglobina umana e canina, tenute a pH neutro e a 4C, mostrano dopo alcuni giorni la formazione di molecole ibride. Nelle stesse condizioni miscele di emoglobine non presentano una formazione significativa di ibridi. I risultati indicano ehe i legami sono più forti nell'emoglobina che nella globina.     

Changes in rate of methemoglobin reduction and oxygen affinity of erythrocytes incubated with inosine,pyruvate and phosphate

Summary Incubation of human erythrocytes with inosine, pyruvate and phosphate increases several fold the ferrihemoglobin reductase activity, the values of which, however, depend on the age of blood (by 6 to 2 times with respect to the normal value of fresh blood).     

Radioprotective action of haematoporphyrin on X-irradiated trypsin

Summary Dose-response curves were obtained for trypsin X-irradiated in aqueous solution (4.6×10^–6M), alone or after incubation with haematoporphyrin (Hp). When trypsin was irradiated alone (exposure doses ranging from 10 to 25 kR) the exponential dose-response curve showed a D₃₇ equal to 17200 R. When trypsin was incubated with Hp before irradiation a D₃₇ equal to 61900 R was obtained, with a dose reduction factor of 3.6. Therefore in our experimental conditions Hp shows a highly significant radioprotective action.Acknowledgments. The authors wish to acknowledge the invaluable suggestions and discussion offered by Prof. G. Cittadini.     

Phoenixin-14: detection and novel physiological implications in cardiac modulation and cardioprotection

Phoenixin-14 (PNX) is a newly identified peptide co-expressed in the hypothalamus with the anorexic and cardioactive Nesfatin-1. Like Nesfatin-1, PNX is able to cross the blood–brain barrier and this suggests a role in peripheral modulation. Preliminary mass spectrography data indicate that, in addition to the hypothalamus, PNX is present in the mammalian heart. This study aimed to quantify PNX expression in the rat heart, and to evaluate whether the peptide influences the myocardial function under basal condition and in the presence of ischemia/reperfusion (I/R). By ELISA the presence of PNX was detected in both hypothalamus and heart. In plasma of normal, but not of obese rats, the peptide concentrations increased after meal. Exposure of the isolated and Langendorff perfused rat heart to exogenous PNX induces a reduction of contractility and relaxation, without effects on coronary pressure and heart rate. As revealed by immunoblotting, these effects were accompanied by an increase of Erk1/2, Akt and eNOS phosphorylation. PNX (EC₅₀ dose), administered after ischemia, induced post-conditioning-like cardioprotection. This was revealed by a smaller infarct size and a better systolic recovery with respect to those detected on hearts exposed to I/R alone. The peptide also activates the cardioprotective RISK and SAFE cascades and inhibits apoptosis. These effects were also observed in the heart of obese rats. Our data provide a first evidence on the peripheral activity of PNX and on its direct cardiomodulatory and cardioprotective role under both normal conditions and in the presence of metabolic disorders.     

Alternative synthetic route for the heterometallic CO-releasing [Sb@Rh12(CO)27]3− icosahedral carbonyl cluster and synthesis of its new unsaturated [Sb@Rh12(CO)24]4− and dimeric [{Sb@Rh12Sb(CO)25}2Rh(CO)2PPh3]7− derivatives

The hetero-metallic [Sb@Rh12(CO)27]3? cluster has been known as for over three decades thanks to Vidal and co-workers, and represents the first example of an E-centered (E=heteroatom) icosahedral rhodium carbonyl cluster. However, its synthesis required high temperature (140–160 °C) and elevated CO pressure (400 atm). Applying the redox condensation method for cluster preparation, we herein report a new synthetic, high-yield route for preparing [Sb@Rh12(CO)27]3? under much milder conditions of temperature and pressure. Notably, when the same synthesis was carried out under N2 instead of CO atmosphere, the new isostructural but unsaturated derivative [Sb@Rh12(CO)24]4? was obtained, for which we report the full X-ray structural characterization. This species represents one of the few examples of an icosahedral cluster disobeying the electron-counting Wade-Mingos rules, possessing less than the expected 170 cluster valence electrons (CVEs). Judging from IR monitoring, the two species can be obtained one from the other by switching between N2 and CO atmosphere, making [Sb@Rh12(CO)27]3? a spontaneous CO-releasing molecule. Finally, the study of the chemical reactivity of [Sb@Rh12(CO)27]3? with PPh3 allowed us to obtain the new [{Sb@Rh12Sb(CO)25}2Rh(CO)2PPh3]7? dimeric compound, for which we herein report the full X-ray structural and 31P NMR analyses.