Genome-Wide Identification of Alternatively Spliced mRNA Targets of Specific RNA-Binding Proteins

BACKGROUND: Alternative splicing plays an important role in generating molecular and functional diversity in multi-cellular organisms. RNA binding proteins play crucial roles in modulating splice site choice. The majority of known binding sites for regulatory proteins are short, degenerate consensus sequences that occur frequently throughout the genome. This poses an important challenge to distinguish between functionally relevant sequences and a vast array of those occurring by chance. METHODOLOGY/PRINCIPAL FINDINGS: Here we have used a computational approach that combines a series of biological constraints to identify uridine-rich sequence motifs that are present within relevant biological contexts and thus are potential targets of the Drosophila master sex-switch protein Sex-lethal (SXL). This strategy led to the identification of one novel target. Moreover, our systematic analysis provides a starting point for the molecular and functional characterization of an additional target, which is dependent on SXL activity, either directly or indirectly, for regulation in a germline-specific manner. CONCLUSIONS/SIGNIFICANCE: This approach has successfully identified previously known, new, and potential SXL targets. Our analysis suggests that only a subset of potential SXL sites are regulated by SXL. Finally, this approach should be directly relevant to the large majority of splicing regulatory proteins for which bonafide targets are unknown

O circuito social da fotografia em Porto Alegre (1922 e 1935)

Photography has been increasingly investigated by Brazilian historians, thus bringing about diverse methodological approaches for the analysis of such visual images. A fundamental component in the studies of this nature is research on photographys social circuit. Within this scope, there is a body of information that is related to the production, circulation and use of photography understood as a visual, creative and disseminating image for certain social imaginaries, and as an artifact that had a place in peoples lives in different contexts. In this article, the author investigates photographys social circuit in the city of Porto Alegre in the 1920s and 1930s, in an attempt to find clues of the presence of this modern invention in the life of Porto Alegre citizens and of the urban vistas that it disseminated

Identification and Characterization of an Unusual Class I Myosin Involved in Vesicle Traffic in Trypanosoma brucei

Myosins are a multimember family of motor proteins with diverse functions in eukaryotic cells. African trypanosomes possess only two candidate myosins and thus represent a useful system for functional analysis of these motors. One of these candidates is an unusual class I myosin (TbMyo1) that is expressed at similar levels but organized differently during the life cycle of Trypanosoma brucei. This myosin localizes to the polarized endocytic pathway in bloodstream forms of the parasite. This organization is actin dependent. Knock down of TbMyo1 results in a significant reduction in endocytic activity, a cessation in cell division and eventually cell death. A striking morphological feature in these cells is an enlargement of the flagellar pocket, which is consistent with an imbalance in traffic to and from the surface. In contrast TbMyo1 is distributed throughout procyclic forms of the tsetse vector and a loss of ∼90% of the protein has no obvious effects on growth or morphology. These results reveal a life cycle stage specific requirement for this myosin in essential endocytic traffic and represent the first description of the involvement of a motor protein in vesicle traffic in these parasites

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Hip-Abductor Fatigue and Single-Leg Landing Mechanics in Women Athletes

CONTEXT: Reduced hip-abductor strength and muscle activation may be associated with altered lower extremity mechanics, which are thought to increase the risk for anterior cruciate ligament injury. However, experimental evidence supporting this relationship is limited. OBJECTIVE: To examine the changes in single-leg landing mechanics and gluteus medius recruitment that occur after a hip-abductor fatigue protocol. DESIGN: Descriptive laboratory study. PATIENTS OR OTHER PARTICIPANTS: Twenty physically active women (age = 21.0 +/- 1.3 years). INTERVENTION(S): Participants were tested before (prefatigue) and after (postfatigue) a hip-abductor fatigue protocol consisting of repetitive side-lying hip abduction. MAIN OUTCOME MEASURE(S): Outcome measures included sagittal-plane and frontal-plane hip and knee kinematics at initial contact and at 60 milliseconds after initial contact during 5 single-leg landings from a height of 40 cm. Peak hip and knee sagittal-plane and frontal-plane joint moments during this time interval were also analyzed. Measures of gluteus medius activation, including latency, peak amplitude, and integrated signal, were recorded. RESULTS: A small (<1 degree) increase in hip-abduction angle at initial contact and a small (<1 degree) decrease in knee-abduction (valgus) angle at 60 milliseconds after contact were observed in the postfatigue landing condition. No other kinematic changes were noted for the knee or hip at initial contact or at 60 milliseconds after initial contact. Peak external knee-adduction moment decreased 27% and peak hip adduction moment decreased 24% during the postfatigue landing condition. Gluteus medius activation was delayed after the protocol, but no difference in peak or integrated signal was seen during the landing trials. CONCLUSIONS: Changes observed during single-leg landings after hip-abductor fatigue were not generally considered unfavorable to the integrity of the anterior cruciate ligament. Further work may be justified to study the role of hip-abductor activation in protecting the knee during landing

Terrestrial and Aquatic Macroinvertebrate Assemblages as a Function of Wetland Type across a Mountain Landscape

Fens and wet meadows are important mountain wetland types, but influences onassemblage structure of associated invertebrates are poorly understood compared with other aspects of the ecology of these habitats. We sought to determine the relative contributions of terrestrial and aquatic invertebrates to diversity and abundance in these wetlands, the extent to which terrestrial and aquatic invertebrate assemblages differ with wetland type, and to what degree the aquatic assemblages vary as a function of slow sheet flow. We compared assemblages in fens and wet meadows, with and without flow, at 80 backcountry sites dispersed across the 6200 km2 landscape of Yosemite, Sequoia,and Kings Canyon National Parks in the Sierra Nevada mountains of California,U.S.A., using standard aquatic and terrestrial sweep netting. Cicadellid leafhoppers, aphids, and thomisid crab spiders were the most abundant terrestrial taxa. Cicadellids,Lepidoptera, anthomyiid, muscid, chloropid, and ephydrid flies, and thomisids were more abundant in fens than in wet meadows. Only mirid leaf bugs were significantly more abundant in wet meadows than fens. Sphaeriid clams and chironomid midges dominated aquatic assemblages both with and without flow. Chloroperlid stoneflies, mites, clams, and flatworms were all more abundant in flow, and Hemiptera and mosquitos were significantly more abundant in quiescent water. Mosquitos were more abundant in wet meadows, but there were few other population differences as a functionof wetland type. Terrestrial diversity was 1.1 to 2.0 times that of aquatic diversity,depending on metric and habitat. Fens had greater terrestrial abundance, richness,evenness, and diversity than wet meadows; there were fewer differences as a function of wetland type for aquatic fauna. Presence or absence of slow sheet flow had more effect on these aquatic assemblages than did wetland type. Cluster analyses, ordination, and multi-response permutation procedures were generally consistent with the univariate results. Vegetation-based wetland classifications should be extrapolated to faunal assemblages with caution, particularly for aquatic invertebrates