Transport proteins determine drug sensitivity and resistance in a protozoan parasite, Trypanosoma brucei

Drug resistance in pathogenic protozoa is very often caused by changes to the ‘transportome’ of the parasites. In Trypanosoma brucei, several transporters have been implicated in uptake of the main classes of drugs, diamidines and melaminophenyl arsenicals. The resistance mechanism had been thought to be due to loss of a transporter known to carry both types of agents: the aminopurine transporter P2, encoded by the gene TbAT1. However, although loss of P2 activity is well-documented as the cause of resistance to the veterinary diamidine diminazene aceturate (Berenil®), cross-resistance between the human-use arsenical melarsoprol and the diamidine pentamidine (MPXR) is the result of loss of a separate High Affinity Pentamidine Transporter (HAPT1). A genome-wide RNAi library screen for resistance to pentamidine, published in 2012, gave the key to the genetic identity of HAPT1 by linking the phenomenon to a locus that contains the closely related T. brucei aquaglyceroporin genes TbAQP2 and TbAQP3. Further analysis determined that knockdown of only one pore, TbAQP2, produced the MPXR phenotype. TbAQP2 is an unconventional aquaglyceroporin with unique residues in the “selectivity region” of the pore, and it was found that in several MPXR lab strains the WT gene was either absent or replaced by a chimeric protein, recombined with parts of TbAQP3. Importantly, wild-type AQP2 was also absent in field isolates of T. b. gambiense, correlating with the outcome of melarsoprol treatment. Expression of a wild-type copy of TbAQP2 in even the most resistant strain completely reversed MPXR and re-introduced HAPT1 function and transport kinetics. Expression of TbAQP2 in Leishmania mexicana introduced a pentamidine transport activity indistinguishable from HAPT1. Although TbAQP2 has been shown to function as a classical aquaglyceroporin it is now clear that it is also a high affinity drug transporter, HAPT1. We discuss here a possible structural rationale for this remarkable ability

Introduzione

L\u2019innovazione e l\u2019internazionalizzazione rappresentano fattori fondamentali del successo economico e competitivo delle imprese. Da un lato il processo di internazionalizzazione risulta favorito dall\u2019innovazione; dall\u2019altro il processo di innovazione appare stimolato dall\u2019entrata in nuovi mercati. Il quadro teorico \ue8 particolarmente articolato e la relazione tra i due fenomeni, sempre pi\uf9 studiata, risulta essere una questione ancora aperta nella letteratura economico-manageriale. I risultati di diversi studi evidenziano infatti l\u2019esistenza di relazioni complesse e la necessit\ue0 di ulteriori approfondimenti. Proprio per cercare di contribuire al dibattito su queste tematiche, \ue8 stato avviato un progetto di ricerca finanziato dall\u2019Universit\ue0 di Trieste (Progetto FRA 2012), di cui questo volume riporta i risultati principali

New VLT observations of the Fermi pulsar PSR J1048-5832

PSR J1048-5832 is a Vela-like (P=123.6 ms; tau~20.3 kyr) gamma-ray pulsar detected by Fermi, at a distance of ~2.7 kpc and with a rotational energy loss rate dot{E}_{SD} ~2 x 10^{36} erg/s. The PSR J1048-5832 field has been observed with the VLT in the V and R bands. We used these data to determine the colour of the object detected closest to the Chandra position (Star D) and confirm that it is not associated with the pulsar. For the estimated extinction along the line of sight, inferred from a re-analysis of the Chandra and XMM-Newton spectra, the fluxes of Star D (V~26.7; R~25.8) imply a -0.13 < (V-R)_0 < 0.6. This means that the PSR J1048-5832 spectrum would be unusually red compared to the Vela pulsar.Moreover, the ratio between the unabsorbed optical and X-ray flux of PSR J1048-5832 would be much higher than for other young pulsars. Thus, we conclude that Star D is not the PSR J1048-5832 counterpart. We compared the derived R and V-band upper limits (R>26.4; V>27.6) with the extrapolation of the X and gamma-ray spectra and constrained the pulsar spectrum at low-energies. In particular, the VLT upper limits suggest that the pulsar spectrum could be consistent with a single power-law, stretching from the gamma-rays to the optical.Comment: 5 pages, 2 figures, accepted for publication on Monthly Notices of the Royal Astronomical Society Main Journa

HST and VLT observations of the neutron star 1E 1207.4-5209

1E 1207.4-5209, the peculiar Central Compact object in the G296.5+10.0 supernova remnant, has been proposed to be an "anti-magnetar" - a young neutron star born with a weak dipole field. Accretion, possibly of supernova fallback material, has also been invoked to explain a large surface temperature anisotropy as well as the generation of peculiar cyclotron absorption features superimposed to its thermal spectrum. Interestingly enough, a faint optical/infrared source was proposed as a possible counterpart to 1E 1207.4-5209, but later questioned, based on coarse positional coincidence. Considering the large offset of 1E 1207.4-5209 with respect to the center of its host supernova remnant, the source should move at ~70 mas/yr. Thus, we tested the association by measuring the proper motion of the proposed optical counterpart. Using HST observations spanning 3.75 years, we computed a 3 sigma upper limit of 7 mas/yr. Absolute astrometry on the same HST data set also places the optical source significantly off the 99% confidence Chandra position. This allows us to safely rule out the association. Using the HST data set, coupled to ground-based observations collected at the ESO/VLT, we set the deepest limits ever obtained to the optical/infrared emission from 1E 1207.4-5209. By combining such limits to the constraints derived from X-ray timing, we rule out accretion as the source of the thermal anisotropy of the neutron star.Comment: 8 pages, 3 figures. Accepted for publication in Astronomy & Astrophysic

Optical Observations of PSR J0205+6449 - the next optical pulsar?

PSR J0205+6449 is a young ({\approx} 5400 years), Crab-like pulsar detected in radio and at X and {\gamma}-ray energies and has the third largest spin-down flux among known rotation powered pulsars. It also powers a bright synchrotron nebula detected in the optical and X-rays. At a distance of {\approx} 3.2 kpc and with an extinction comparable to the Crab, PSR J0205+6449 is an obvious target for optical observations. We observed PSR J0205+6449 with several optical facilities, including 8m class ground-based telescopes, such as the Gemini and the Gran Telescopio Canarias. We detected a point source, at a significance of 5.5{\sigma}, of magnitude i {\approx} 25.5, at the centre of the optical synchrotron nebula, coincident with the very accurate Chandra and radio positions of the pulsar. Thus, we discovered a candidate optical counterpart to PSR J0205+6449. The pulsar candidate counterpart is also detected in the g ({\approx}27.4) band and weakly in the r ({\approx}26.2) band. Its optical spectrum is fit by a power law with photon index {\Gamma}0 = 1.9{\pm}0.5, proving that the optical emission if of non-thermal origin, is as expected for a young pulsar. The optical photon index is similar to the X-ray one ({\Gamma}X = 1.77{\pm}0.03), although the optical fluxes are below the extrapolation of the X-ray power spectrum. This would indicate the presence of a double spectral break between the X-ray and optical energy range, at variance with what is observed for the Crab and Vela pulsars, but similar to the Large Magellanic Cloud pulsar PSR B0540-69.Comment: 13 Pages, 4 Tables, 7 Figures, Accepted for publication in MNRA