Effect of stress and diapause in two Calliphoridae species

Cultures of two Dipteran flies (Calliphora vicina (R-D) and C. vomitoria (L.)) were established to answer questions in regards to responses to thermal and desiccation stress, effects of diapause and the mechanisms which underpin diapause. The findings are divided in to two sections. Unequivocal new findings – Calliphora vomitoria were seen to depend on water being present in culture medium for increased survival. Furthermore, C. vomitoria were found to have lower desiccation resistance than C. vicina. Larvae of C. vicina and C. vomitoria showed different cold tolerance strategies, with C. vicina being freeze-avoiding and C. vomitoria ‘partially’ freeze-tolerant. Metabolomics, using $^1$H-NMR, revealed that diapause and non-diapause had distinct metabolic profiles. Diapause larvae were seen to reduce energy synthesis from the Krebs cycle and increase glycolysis. Calliphora vicina and C. vomitoria also exhibited different diapause phenotypes; C. vicina entered a maternally regulated facultative diapause as an L3 larvae, Calliphora vomitoria had a less distinct diapause, with maternal conditions having little effect. Speculative new findings - Despite the above differences C. vicina and C. vomitoria were able to produce a viable cross, though field fresh C. vomitoria were not used, as such it cannot be confirmed if this could occur in the wild. Increased temperatures due to climate change may affect both phenology and survival of insects; C. vicina was seen to have a delayed induction to diapause and a reduction in the proportion entering diapause. Diapause conferred increased cold tolerance; therefore those insects that overwinter not in diapause may suffer increased mortality

Do media literacies approach equity and justice?

It is often assumed that media literacy serves to protect and uphold democratic practice and that media literate citizens are the best safeguards for democracy. However, little attention is paid to defining this practice and its relationship to ongoing inequities within democratic societies. In this essay, we argue media literacy operates from three core assumptions; media literacy creates knowledgeable individuals, empowers communities, and encourages democratic participation. The first assumption draws out an individual’s skills and critical thinking in media literacy practices. The second assumption focuses on the community aspect of media literacy, specifically which communities are best served by media literacy and why. Finally, the connection between media literacy and democratic practices is evaluated to understand how the democratic ideals of equity and justice are situated within the existing literature. Through an exploration of these assumptions, this essay provokes a discussion into the assumptions that media literacy scholarship and practice addresses to highlight some of the gaps in constructing impactful practice that centers on equity and social justice

Do Media Literacies Approach Equity and Justice?

It is often assumed that media literacy serves to protect and uphold democratic practice and that media literate citizens are the best safeguards for democracy. However, little attention is paid to defining this practice and its relationship to ongoing inequities within democratic societies. In this essay, we argue media literacy operates from three core assumptions; media literacy creates knowledgeable individuals, empowers communities, and encourages democratic participation. The first assumption draws out an individual’s skills and critical thinking in media literacy practices. The second assumption focuses on the community aspect of media literacy, specifically which communities are best served by media literacy and why. Finally, the connection between media literacy and democratic practices is evaluated to understand how the democratic ideals of equity and justice are situated within the existing literature. Through an exploration of these assumptions, this essay provokes a discussion into the assumptions that media literacy scholarship and practice addresses to highlight some of the gaps in constructing impactful practice that centers on equity and social justice

MPLEx: a Robust and Universal Protocol for Single-Sample Integrative Proteomic, Metabolomic, and Lipidomic Analyses

ABSTRACT Integrative multi-omics analyses can empower more effective investigation and complete understanding of complex biological systems. Despite recent advances in a range of omics analyses, multi-omic measurements of the same sample are still challenging and current methods have not been well evaluated in terms of reproducibility and broad applicability. Here we adapted a solvent-based method, widely applied for extracting lipids and metabolites, to add proteomics to mass spectrometry-based multi-omics measurements. The m etabolite, p rotein, and l ipid ex traction (MPLEx) protocol proved to be robust and applicable to a diverse set of sample types, including cell cultures, microbial communities, and tissues. To illustrate the utility of this protocol, an integrative multi-omics analysis was performed using a lung epithelial cell line infected with Middle East respiratory syndrome coronavirus, which showed the impact of this virus on the host glycolytic pathway and also suggested a role for lipids during infection. The MPLEx method is a simple, fast, and robust protocol that can be applied for integrative multi-omic measurements from diverse sample types (e.g., environmental, in vitro , and clinical). IMPORTANCE In systems biology studies, the integration of multiple omics measurements (i.e., genomics, transcriptomics, proteomics, metabolomics, and lipidomics) has been shown to provide a more complete and informative view of biological pathways. Thus, the prospect of extracting different types of molecules (e.g., DNAs, RNAs, proteins, and metabolites) and performing multiple omics measurements on single samples is very attractive, but such studies are challenging due to the fact that the extraction conditions differ according to the molecule type. Here, we adapted an organic solvent-based extraction method that demonstrated broad applicability and robustness, which enabled comprehensive proteomics, metabolomics, and lipidomics analyses from the same sample

MPLEx: a method for simultaneous pathogen inactivation and extraction of samples for multi-omics profiling

The continued emergence and spread of infectious agents is of great concern, and systems biology approaches to infectious disease research can advance our understanding of host–pathogen relationships and facilitate the development of new therapies and vaccines

Muscle Segment Homeobox Genes Direct Embryonic Diapause by Limiting Inflammation in the Uterus

Embryonic diapause is a reproductive strategy widespread in the animal kingdom. This phenomenon is defined by a temporary arrest in blastocyst growth and metabolic activity within a quiescent uterus without implantation until the environmental and maternal milieu become favorable for pregnancy to progress. We found that uterine Msx expression persists during diapause across species; their inactivation in the mouse uterus results in termination of diapause with the development of implantation-like responses (“pseudoimplantation”) that ultimately succumbed to resorption. To understand the cause of this failure, we compared proteome profiles between floxed and Msx-deleted uteri. In deleted uteri, several functional networks, including transcription/translation, ubiquitin-proteasome, inflammation, and endoplasmic reticulum stress, were dysregulated. Computational modeling predicted intersection of these pathways on an enhanced inflammatory signature. Further studies showed that this signature was reflected in increased phosphorylated IκB levels and nuclear NFκB in deleted uteri. This was associated with enhanced proteasome activity and endoplasmic reticulum stress. Interestingly, treatment with anti-inflammatory glucocorticoid (dexamethasone) reduced the inflammatory signature with improvement of the diapause phenotype. These findings highlight an unexpected role of uterine Msx in limiting aberrant inflammatory responses to maintain embryonic diapause

MERS-CoV and H5N1 influenza virus antagonize antigen presentation by altering the epigenetic landscape

Convergent evolution dictates that diverse groups of viruses will target both similar and distinct host pathways to manipulate the immune response and improve infection. In this study, we sought to leverage this uneven viral antagonism to identify critical host factors that govern disease outcome. Utilizing a systems-based approach, we examined differential regulation of IFN-γ–dependent genes following infection with robust respiratory viruses including influenza viruses [A/influenza/Vietnam/1203/2004 (H5N1-VN1203) and A/influenza/California/04/2009 (H1N1-CA04)] and coronaviruses [severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome CoV (MERS-CoV)]. Categorizing by function, we observed down-regulation of gene expression associated with antigen presentation following both H5N1-VN1203 and MERS-CoV infection. Further examination revealed global down-regulation of antigen-presentation gene expression, which was confirmed by proteomics for both H5N1-VN1203 and MERS-CoV infection. Importantly, epigenetic analysis suggested that DNA methylation, rather than histone modification, plays a crucial role in MERS-CoV–mediated antagonism of antigen-presentation gene expression; in contrast, H5N1-VN1203 likely utilizes a combination of epigenetic mechanisms to target antigen presentation. Together, the results indicate a common mechanism utilized by H5N1-VN1203 and MERS-CoV to modulate antigen presentation and the host adaptive immune response

Adding 6 months of androgen deprivation therapy to postoperative radiotherapy for prostate cancer: a comparison of short-course versus no androgen deprivation therapy in the RADICALS-HD randomised controlled trial

Background Previous evidence indicates that adjuvant, short-course androgen deprivation therapy (ADT) improves metastasis-free survival when given with primary radiotherapy for intermediate-risk and high-risk localised prostate cancer. However, the value of ADT with postoperative radiotherapy after radical prostatectomy is unclear. Methods RADICALS-HD was an international randomised controlled trial to test the efficacy of ADT used in combination with postoperative radiotherapy for prostate cancer. Key eligibility criteria were indication for radiotherapy after radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to radiotherapy alone (no ADT) or radiotherapy with 6 months of ADT (short-course ADT), using monthly subcutaneous gonadotropin-releasing hormone analogue injections, daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as distant metastasis arising from prostate cancer or death from any cause. Standard survival analysis methods were used, accounting for randomisation stratification factors. The trial had 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 80% to 86% (hazard ratio [HR] 0·67). Analyses followed the intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov, NCT00541047. Findings Between Nov 22, 2007, and June 29, 2015, 1480 patients (median age 66 years [IQR 61–69]) were randomly assigned to receive no ADT (n=737) or short-course ADT (n=743) in addition to postoperative radiotherapy at 121 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 9·0 years (IQR 7·1–10·1), metastasis-free survival events were reported for 268 participants (142 in the no ADT group and 126 in the short-course ADT group; HR 0·886 [95% CI 0·688–1·140], p=0·35). 10-year metastasis-free survival was 79·2% (95% CI 75·4–82·5) in the no ADT group and 80·4% (76·6–83·6) in the short-course ADT group. Toxicity of grade 3 or higher was reported for 121 (17%) of 737 participants in the no ADT group and 100 (14%) of 743 in the short-course ADT group (p=0·15), with no treatment-related deaths. Interpretation Metastatic disease is uncommon following postoperative bed radiotherapy after radical prostatectomy. Adding 6 months of ADT to this radiotherapy did not improve metastasis-free survival compared with no ADT. These findings do not support the use of short-course ADT with postoperative radiotherapy in this patient population