Norma Gorrochotegui

Secondary bacterial infection (superinfection) post influenza is a serious clinical complication often leading to pneumonia and death. Eicosanoids are bioactive lipid mediators that play critical roles in the induction and resolution of inflammation. CYP450 lipid metabolites are anti-inflammatory lipid mediators that are produced at an excessive level during superinfection potentiating the vulnerability to secondary bacterial infection. Using Nanostring nCounter technology, we have defined the targeted transcriptional response where CYP450 metabolites dampen the Toll-like receptor signaling in macrophages. CYP450 metabolites are endogenous ligands for the nuclear receptor and transcription factor, PPARα. Activation of PPARα hinders NFκB p65 activities by altering its phosphorylation and nuclear translocation during TLR stimulation. Additionally, activation of PPARα inhibited anti-bacterial activities and enhanced macrophage polarization to an anti-inflammatory subtype (M2b). Lastly,...

Understanding the processes by which species colonize and adapt to human habitats is particularly important in the case of disease-vectoring arthropods. The mosquito species Aedes aegypti, a major vector of dengue and yellow fever viruses, probably originated as a wild, zoophilic species in sub-Saharan Africa, where some populations still breed in tree holes in forested habitats. Many populations of the species, however, have evolved to thrive in human habitats and to bite humans. This includes some populations within Africa as well as almost all those outside Africa. It is not clear whether all domestic populations are genetically related and represent a single ‘domestication’ event, or whether association with human habitats has developed multiple times independently within the species. To test the hypotheses above, we screened 24 worldwide population samples of Ae. aegypti at 12 polymorphic microsatellite loci. We identified two distinct genetic clusters: one included all domestic populations outside of Africa and the other included both domestic and forest populations within Africa. This suggests that human association in Africa occurred independently from that in domestic populations across the rest of the world. Additionally, measures of genetic diversity support Ae. aegypti in Africa as the ancestral form of the species. Individuals from domestic populations outside Africa can reliably be assigned back to their population of origin, which will help determine the origins of new introductions of Ae. aegypti

Historically, it has been difficult to perform gene knockouts on a large scale in Neurospora, due to the low frequency of homologous recombination. For a functional genomics project, we developed a feasible high-throughput approach that has two significant components: 1) Yeast recombinational cloning was used to create the knockout cassettes; and 2) Novel mus-51 and mus-52 mutant strains based on the work of Inoue and coworkers were used as the recipients to virtually eliminate ectopic insertion of the knockout cassettes. The process of generating cassettes was performed on a pipetting robot but the essentials of the procedure were developed manually and are presented here. The overall scheme and the details for primer design are described in Colot and Park et al. The creation and characterization of the Neurospora knockout strains incorporate several modifications to existing techniques to facilitate handling of large numbers of strains, as well as an efficient and convenient metho...

Understanding the processes by which species colonize and adapt to human habitats is particularly important in the case of disease-vectoring arthropods. The mosquito speciesAedes aegypti, a major vector of dengue and yellow fever viruses, probably originated as a wild, zoophilic species in sub-Saharan Africa, where some populations still breed in tree holes in forested habitats. Many populations of the species, however, have evolved to thrive in human habitats and to bite humans. This includes some populations within Africa as well as almost all those outside Africa. It is not clear whether all domestic populations are genetically related and represent a single ‘domestication’ event, or whether association with human habitats has developed multiple times independently within the species. To test the hypotheses above, we screened 24 worldwide population samples ofAe. aegyptiat 12 polymorphic microsatellite loci. We identified two distinct genetic clusters: one included all domestic p...

Patients infected with influenza are at high risk of secondary bacterial infection, which is a major proximate cause of morbidity and mortality. We have shown that in mice, prior infection with influenza results in increased inflammation and mortality upon Staphylococcus aureus infection, recapitulating the human disease. Lipidomic profiling of the lungs of superinfected mice revealed an increase in CYP450 metabolites during lethal superinfection. These lipids are endogenous ligands for the nuclear receptor PPARα, and we demonstrate that Ppara−/− mice are less susceptible to superinfection than wild-type mice. PPARα is an inhibitor of NFκB activation, and transcriptional profiling of cells isolated by bronchoalveolar lavage confirmed that influenza infection inhibits NFκB, thereby dampening proinflammatory and prosurvival signals. Furthermore, network analysis indicated an increase in necrotic cell death in the lungs of superinfected mice compared to mice infected with S. aureus alo...

Field trials of the predacious copepod Mesocyclops longisetus Thiubaud were conducted in northeastern Mexico to determine the effectiveness of this species to control larval Aedes aegypti (L.) populations and to survive and reproduce in nature. Groups of 200, 50, and 50 ovigerous M. longisetus females were inoculated into 200-liter metal drums, discarded tires, and cemetery flower vases, respectively, which are 3 of the more important Aedes breeding sites in this area. Larvae were sampled at 15-d intervals, and total surviving cyclops were collected at the end of the study, 120 d later. Community participation was solicited through a simple training program on copepod rescue before drum cleaning and facilitated by the addition of a drum marker to remind residents of copepod presence. Results showed good cooperation and after 4 mo all peridomestic drums, still supported variable numbers of cyclopoids. Average of larvae reduction was 37.5% for drums, 67.5% for flower vases, and 40.9% for tires. This study shows difficulties of using cyclopoids for tires and vases in areas where prolonged dry seasons desiccated these habitats and reduced copepod survival.