Differential Gel Electrophoresis (DIGE) Evaluation of Naphthoimidazoles Mode of Action: A Study in Trypanosoma cruzi Bloodstream Trypomastigotes

BackgroundThe obligate intracellular protozoan Trypanosoma cruzi is the causative agent of Chagas disease, a neglected illness affecting millions of people in Latin America that recently entered non-endemic countries through immigration, as a consequence of globalization. The chemotherapy for this disease is based mainly on benznidazole and nifurtimox, which are very efficient nitroderivatives against the acute stage but present limited efficacy during the chronic phase. Our group has been studying the trypanocidal effects of naturally occurring quinones and their derivatives, and naphthoimidazoles derived from β-lapachone N1, N2 and N3 were the most active. To assess the molecular mechanisms of action of these compounds, we applied proteomic techniques to analyze treated bloodstream trypomastigotes, which are the clinically relevant stage of the parasite.Conclusion/SignificanceOur results point to different modes of action for N1, N2 and N3, which indicate a great variety of metabolic pathways involved and allow for novel perspectives on the development of trypanocidal agents.     

FAS haploinsufficiency is a common disease mechanism in the human autoimmune lymphoproliferative syndrome

The autoimmune lymphoproliferative syndrome (ALPS) is characterized by early-onset lymphadenopathy, splenomegaly, immune cytopenias, and an increased risk for B cell lymphomas. Most ALPS patients harbor mutations in the FAS gene, which regulates lymphocyte apoptosis. These are commonly missense mutations affecting the intracellular region of the protein and have a dominant-negative effect on the signaling pathway. However, analysis of a large cohort of ALPS patients revealed that ～30% have mutations affecting the extracellular region of FAS, and among these, 70% are nonsense, splice site, or insertions/deletions with frameshift for which no dominant-negative effect would be expected. We evaluated the latter patients to understand the mechanism(s) by which these mutations disrupted the FAS pathway and resulted in clinical disease. We demonstrated that most extracellular-region FAS mutations induce low FAS expression due to nonsense-mediated RNA decay or protein instability, resulting in defective death-inducing signaling complex formation and impaired apoptosis, although to a lesser extent as compared with intracellular mutations. The apoptosis defect could be corrected by FAS overexpression in vitro. Our findings define haploinsufficiency as a common disease mechanism in ALPS patients with extracellular FAS mutations.     

New southerly breeding location of king penguins (Aptenodytes patagonicus) on Elephant Island (Maritime Antarctic)

In the 2009–2010 austral summer, two breeding pairs of king penguins were recorded at Stinker Point, Elephant Island, Maritime Antarctic. This is the first record of king penguins breeding south of 60°S. The finding suggests a possible range extension of this species and increases the number of breeding bird species at Stinker Point, which was recently appointed as an Important Bird Area in Antarctica.     

Validation of the determination of oxymetholone in human plasma analysis using gas chromatography-mass spectrometry. Application to pharmacokinetic studies

A simple and rapid procedure for extraction of oxymetholone in human plasma using gas chromatography coupled with quadrupole mass spectrometric was evaluated. The method involves analyte extraction with tert.-butylmethylether after alkalinization of the plasma and derivatization with MSTFA-NH(4)I-2-mercaptoethanol before the high resolution gas chromatographic-mass spectrometry separation. Methyltestosterone was used as internal standard. The calibration curves were linear, with typical r(2) values >0.995 and F(table)>F(calculated) (alpha=0.05). Recovery from plasma proved to be more than 70%. The method was accurate and reproducible, and was successfully applied to determine the pharmacokinetic parameters of oxymetholone for healthy volunteers after oral administration of 50 mg of the compound. The (C(max)) and (T(max)) were 18.8 +/- 0.4 ng/ml and 210 +/- 42.4 min, respectively.     

Breeding sites and population of seabirds on Admiralty Bay,King George Island,Antarctica

The monitoring of the status and distribution of seabird populations is necessary to understand their spatial and temporal responses to rapid climate changes occurring in the Western Antarctic Peninsula area. We surveyed and mapped Admiralty Bay bird communities and related them to climate variables—temperature, temperature anomaly, Antarctic Oscillation Index and El-Niño Southern Oscillation Index. We recorded 13 breeding seabird species over three seasons (2009/2010, 2010/2011 and 2011/2012) and mapped 10 of them over an area of 149.5 ha. The ice-free areas with the greatest number of species were Point Thomas, Keller Peninsula and Hennequin Point. The most abundant species was the Adelie Penguin (Pygoscelis adeliae) followed by the Chinstrap Penguin (P. antarcticus). We observed that the number of breeding pairs of Gentoo Penguins (P. papua), Chinstrap Penguins and skuas (Catharacta maccormicki and C. antarctica) are related to temperature, temperature anomaly and El-Niño Southern Oscillation Index. The size of breeding populations and their distributions have been fluctuating over the last 30 years in ice-free areas of Admiralty Bay. Most species showed a decreasing trend from 1978 to 2012, with the exception of Chinstrap Penguins, Southern Giant Petrels (Macronectes giganteus) and skuas, which seem to be stable in numbers in the last two decades. Decreases in seabird populations from the Antarctic Peninsula are widely recognized as a response to environmental change and anthropogenic influences such as tourism and building activities, thus highlighting the importance of monitoring to support mitigation measures.