Haunted Mirror: British Gothic Masculinity in Transatlantic Cinema

This project follows the ghost of Gothic British masculinity across the ocean in the period of classical cinema. It examines the ways in which British stars were offered as an alternative to the American ideal of muscular, anti-intellectual, tough male identity. British men on film allowed Hollywood a glimpse in a mirror, a dark, haunted mirror, where identity might be fractured, damaged, liberated, queered or feminised. In a period dominated by two world wars and a Great Depression, identities of all types were being challenged and filmmakers used Britishness to allow this tension to seep into cinema. This project uses the lens of the Gothic as a method of uncovering the hidden history that is embedded in many films. The uncanny and the sublime, shadows and mirrors, portraits, decadent iconography and dark doubles all dominate in these cinematic texts. At a time when the Production Code made it necessary for subversive content to be well hidden, films contained embedded secret codes and invited possible alternative readings. Bringing together film scholarship with literary theorists this thesis offers fresh perspectives on historical cinematic meanings. This study presents a detailed analysis of British male stardom as it emerged in the period of early talkies. It details the ways in which the male stars, Ronald Colman, Basil Rathbone and George Sanders were presented in fan publications. It presents the contradictions inherent in their fan discourse and allows for consideration of the queerness that American culture seemed to accept was part of British - and European - male identity

Exploring access to community neurorehabilitation for people with progressive neurological conditions: a qualitative study

Purpose: Community neurorehabilitation enables people with progressive neurological conditions (PNCs) to manage their symptoms to live an active, fulfilling life; however, it is not accessible to all. This study explored the factors influencing access to community neurorehabilitation in Northern Ireland from the perspective of people with PNCs and their carers. Methods: Eleven people living with a PNC and three carers took part in virtual focus groups. Data was thematically analysed using the framework method. Results: Access to neurorehabilitation was described as a staged journey, driven by people with PNCs, and impacted by interactions with others. Four themes were identified: the person in the driving seat, describing the value of person-centred care and the need for proactivity; the traffic lights, depicting the role and influence of health care professionals (HCPs); the need for direction; and roadworks and roadblocks, identifying additional barriers to access. In addition, six fundamentals of good access were identified. Conclusions: This study adds depth to our understanding of the complexity, and the roles and needs of people with PNCs and HCPs, in accessing community neurorehabilitation. Further research is needed to determine how best to empower people to access rehabilitation.</p

A Thioredoxin Domain-Containing Protein Interacts with Pepino mosaic virus Triple Gene Block Protein 1

Pepino mosaic virus (PepMV) is a mechanically-transmitted tomato pathogen of importance worldwide. Interactions between the PepMV coat protein and triple gene block protein (TGBp1) with the host heat shock cognate protein 70 and catalase 1 (CAT1), respectively, have been previously reported by our lab. In this study, a novel tomato interactor (SlTXND9) was shown to bind the PepMV TGBp1 in yeast-two-hybrid screening, in vitro pull-down and bimolecular fluorescent complementation (BiFC) assays. SlTXND9 possesses part of the conserved thioredoxin (TRX) active site sequence (W__PC vs. WCXPC), and TXND9 orthologues cluster within the TRX phylogenetic superfamilyclosesttophosducin-likeprotein-3. InPepMV-infectedandhealthyNicotianabenthamiana plants,NbTXND9mRNAlevelswerecomparable,andexpressionlevelsremainedstableinbothlocal and systemic leaves for 10 days post inoculation (dpi), as was also the case for catalase 1 (CAT1). To localize the TXND9 in plant cells, a polyclonal antiserum was produced. Purified α-SlTXND9 immunoglobulin (IgG) consistently detected a set of three protein bands in the range of 27–35 kDa, in the 1000 and 30,000 g pellets, and the soluble fraction of extracts of healthy and PepMV-infected N. benthamiana leaves, but not in the cell wall. These bands likely consist of the homologous protein NbTXND9 and its post-translationally modified derivatives. On electron microscopy, immuno-gold labellingofultrathinsectionsofPepMV-infectedN.benthamianaleavesusingα-SlTXND9IgGrevealed particle accumulation close to plasmodesmata, suggesting a role in virus movement. Taken together, this study highlights a novel tomato-PepMV protein interaction and provides data on its localization in planta. Currently, studies focusing on the biological function of this interaction during PepMV infection are in progress

Familial myelodysplasia and acute myeloid leukaemia--a review

Summary Familial occurrence of myelodysplasia (MDS) and/or acute myeloid leukaemia (AML) is rare but can provide a useful resource for the investigation of predisposing mutations in these myeloid malignancies. To date, examination of families with MDS/AML has lead to the detection of two culprit genes, RUNX1 and CEBPA. Germline mutations in RUNX1 result in familial platelet disorder with propensity to myeloid malignancy and inherited mutations of CEBPA predispose to AML. Unfortunately, the genetic cause remains obscure in most other reported pedigrees. Further insight into the molecular mechanisms of familial MDS/AML will require awareness by clinicians of new patients with relevant family histories

Cucurbit chlorotic yellows virus p22 suppressor of RNA silencing binds single-, double-stranded long and short interfering RNA molecules in vitro

[EN] Cucurbit chlorotic yellows virus (CCYV) is a new member of the genus Crinivirus (family Closteroviridae) with a bi-partite genome. CCYV RNA 1-encoded p22 has recently been reported to be a weak local suppressor of RNA silencing for which an interaction with cucumber SKP1LB1 through an F-box-like motif was demonstrated to be essential. Using a bacterially expressed maltose-binding protein (MBP) fusion of CCYV p22 in electrophoretic mobility shift assays (EMSA), we have examined in vitro its ability to bind different RNA templates. Our experiments showed that CCYV p22 is able to bind to ss and ds long RNAs, in addition to ss and ds small interfering (si) RNA molecules. CCYV p22 deletion mutants (MBP_CCYV DEL1-4) were produced that covered the entire protein, with MBP_CCYV DEL2 corresponding to the F-box motif and its flanking sequences. None of these deletions abolished the capacity of CCYV p22 to bind ss- and dsRNA molecules. However, deletions affecting the C-terminal half of the protein resulted in decreased binding efficiency for either ss- or dsRNA molecules indicating that essential elements for these interactions are located in this region. Taken together, our data add to current knowledge of the mode of action of suppressors of RNA silencing encoded by genes sited at the 3'-terminus of crinivirus genomic RNA 1, and shed light on the involvement of CCYV p22 in the suppression of RNA silencing and/or in another role in the virus life cycle via RNA binding.Salavert, F.; Navarro Bohigues, JA.; Owen, CA.; Khechmar, S.; Pallás Benet, V.; Livieratos, IC. (2020). Cucurbit chlorotic yellows virus p22 suppressor of RNA silencing binds single-, double-stranded long and short interfering RNA molecules in vitro. Virus Research. 279:1-8. https://doi.org/10.1016/j.virusres.2020.197887S18279Abrahamian, P. E., Seblani, R., Sobh, H., & Abou-Jawdah, Y. (2013). Detection and quantitation of two cucurbit criniviruses in mixed infection by real-time RT-PCR. Journal of Virological Methods, 193(2), 320-326. doi:10.1016/j.jviromet.2013.06.004Bananej, K., Menzel, W., Kianfar, N., Vahdat, A., & Winter, S. (2013). First Report of Cucurbit chlorotic yellows virus Infecting Cucumber, Melon, and Squash in Iran. Plant Disease, 97(7), 1005-1005. doi:10.1094/pdis-01-13-0125-pdnBrantley, J. D., & Hunt, A. G. (1993). The N-terminal protein of the polyprotein encoded by the potyvirus tobacco vein mottling virus is an RNA-binding protein. Journal of General Virology, 74(6), 1157-1162. doi:10.1099/0022-1317-74-6-1157BUCK, K. W., & KEMPSON-JONES, G. F. (1970). Three Types of Virus Particle in Penicillium stoloniferum. Nature, 225(5236), 945-946. doi:10.1038/225945a0Cañizares, M. C., Navas-Castillo, J., & Moriones, E. (2008). Multiple suppressors of RNA silencing encoded by both genomic RNAs of the crinivirus, Tomato chlorosis virus. Virology, 379(1), 168-174. doi:10.1016/j.virol.2008.06.020Chen, S., Sun, X., Shi, Y., Wei, Y., Han, X., Li, H., … Shi, Y. (2019). Cucurbit Chlorotic Yellows Virus p22 Protein Interacts with Cucumber SKP1LB1 and Its F-Box-Like Motif Is Crucial for Silencing Suppressor Activity. Viruses, 11(9), 818. doi:10.3390/v11090818Cuellar, W. J., Tairo, F., Kreuze, J. F., & Valkonen, J. P. T. (2008). Analysis of gene content in sweet potato chlorotic stunt virus RNA1 reveals the presence of the p22 RNA silencing suppressor in only a few isolates: implications for viral evolution and synergism. Journal of General Virology, 89(2), 573-582. doi:10.1099/vir.0.83471-0Cuellar, W. J., Kreuze, J. F., Rajamaki, M.-L., Cruzado, K. R., Untiveros, M., & Valkonen, J. P. T. (2009). Elimination of antiviral defense by viral RNase III. Proceedings of the National Academy of Sciences, 106(25), 10354-10358. doi:10.1073/pnas.0806042106GYOUTOKU, Y., OKAZAKI, S., FURUTA, A., ETOH, T., MIZOBE, M., KUNO, K., … OKUDA, M. (2009). Chlorotic yellows disease of melon caused by Cucurbit chlorotic yellows virus, a new crinivirus. Japanese Journal of Phytopathology, 75(2), 109-111. doi:10.3186/jjphytopath.75.109Herranz, M. C., & Pallás, V. (2004). RNA-binding properties and mapping of the RNA-binding domain from the movement protein of Prunus necrotic ringspot virus. Journal of General Virology, 85(3), 761-768. doi:10.1099/vir.0.19534-0Huang, L.-H., Tseng, H.-H., Li, J.-T., & Chen, T.-C. (2010). First Report of Cucurbit chlorotic yellows virus Infecting Cucurbits in Taiwan. Plant Disease, 94(9), 1168-1168. doi:10.1094/pdis-94-9-1168bKataya, A. R. A., Suliman, M. N. S., Kalantidis, K., & Livieratos, I. C. (2009). Cucurbit yellow stunting disorder virus p25 is a suppressor of post-transcriptional gene silencing. Virus Research, 145(1), 48-53. doi:10.1016/j.virusres.2009.06.010Klaassen, V. A., Mayhew, D., Fisher, D., & Falk, B. W. (1996). In VitroTranscripts from Cloned cDNAs of the Lettuce Infectious Yellows Closterovirus Bipartite Genomic RNAs Are Competent for Replication inNicotiana benthamianaProtoplasts. Virology, 222(1), 169-175. doi:10.1006/viro.1996.0407Kreuze, J. F., Savenkov, E. I., Cuellar, W., Li, X., & Valkonen, J. P. T. (2005). Viral Class 1 RNase III Involved in Suppression of RNA Silencing. Journal of Virology, 79(11), 7227-7238. doi:10.1128/jvi.79.11.7227-7238.2005Kubota, K., & Ng, J. C. K. (2016). Lettuce chlorosis virus P23 Suppresses RNA Silencing and Induces Local Necrosis with Increased Severity at Raised Temperatures. Phytopathology®, 106(6), 653-662. doi:10.1094/phyto-09-15-0219-rLandeo-Ríos, Y., Navas-Castillo, J., Moriones, E., & Cañizares, M. C. (2016). The p22 RNA silencing suppressor of the crinivirus Tomato chlorosis virus preferentially binds long dsRNAs preventing them from cleavage. Virology, 488, 129-136. doi:10.1016/j.virol.2015.11.008Marcos, J. F., Vilar, M., Pérez-Payá, E., & Pallás, V. (1999). In VivoDetection, RNA-Binding Properties and Characterization of the RNA-Binding Domain of the p7 Putative Movement Protein from Carnation Mottle Carmovirus (CarMV). Virology, 255(2), 354-365. doi:10.1006/viro.1998.9596Mashiko, T., Wang, W.-Q., Hartono, S., Suastica, G., Neriya, Y., Nishigawa, H., & Natsuaki, T. (2019). The p27 open reading frame of tomato infectious chlorosis virus encodes a suppressor of RNA silencing. Journal of General Plant Pathology, 85(4), 301-305. doi:10.1007/s10327-019-00850-0Navarro, J. A., Genovés, A., Climent, J., Saurí, A., Martínez-Gil, L., Mingarro, I., & Pallás, V. (2006). RNA-binding properties and membrane insertion of Melon necrotic spot virus (MNSV) double gene block movement proteins. Virology, 356(1-2), 57-67. doi:10.1016/j.virol.2006.07.040Okuda, M., Okazaki, S., Yamasaki, S., Okuda, S., & Sugiyama, M. (2010). Host Range and Complete Genome Sequence of Cucurbit chlorotic yellows virus, a New Member of the Genus Crinivirus. Phytopathology®, 100(6), 560-566. doi:10.1094/phyto-100-6-0560Orfanidou, C., Maliogka, V. I., & Katis, N. I. (2014). First Report of Cucurbit chlorotic yellows virus in Cucumber, Melon, and Watermelon in Greece. Plant Disease, 98(10), 1446-1446. doi:10.1094/pdis-03-14-0311-pdnOrfanidou, C. G., Mathioudakis, M. M., Katsarou, K., Livieratos, I., Katis, N., & Maliogka, V. I. (2019). Cucurbit chlorotic yellows virus p22 is a suppressor of local RNA silencing. Archives of Virology, 164(11), 2747-2759. doi:10.1007/s00705-019-04391-xOrílio, A. F., Fortes, I. M., & Navas-Castillo, J. (2014). Infectious cDNA clones of the crinivirus Tomato chlorosis virus are competent for systemic plant infection and whitefly-transmission. Virology, 464-465, 365-374. doi:10.1016/j.virol.2014.07.032Pumplin, N., & Voinnet, O. (2013). RNA silencing suppression by plant pathogens: defence, counter-defence and counter-counter-defence. Nature Reviews Microbiology, 11(11), 745-760. doi:10.1038/nrmicro3120Richmond, K. E., Chenault, K., Sherwood, J. L., & German, T. L. (1998). Characterization of the Nucleic Acid Binding Properties of Tomato Spotted Wilt Virus Nucleocapsid Protein. Virology, 248(1), 6-11. doi:10.1006/viro.1998.9223Salem, N. M., Chen, A. Y. S., Tzanetakis, I. E., Mongkolsiriwattana, C., & Ng, J. C. K. (2009). Further complexity of the genus Crinivirus revealed by the complete genome sequence of Lettuce chlorosis virus (LCV) and the similar temporal accumulation of LCV genomic RNAs 1 and 2. Virology, 390(1), 45-55. doi:10.1016/j.virol.2009.04.025Serra-Soriano, M., Antonio Navarro, J., & Pallás, V. (2016). Dissecting the multifunctional role of the N-terminal domain of theMelon necrotic spot viruscoat protein in RNA packaging, viral movement and interference with antiviral plant defence. Molecular Plant Pathology, 18(6), 837-849. doi:10.1111/mpp.12448Simon, A. E., & Miller, W. A. (2013). 3′ Cap-Independent Translation Enhancers of Plant Viruses. Annual Review of Microbiology, 67(1), 21-42. doi:10.1146/annurev-micro-092412-155609Voinnet, O. (2005). Induction and suppression of RNA silencing: insights from viral infections. Nature Reviews Genetics, 6(3), 206-220. doi:10.1038/nrg1555Wang, J., Stewart, L. R., Kiss, Z., & Falk, B. W. (2010). Lettuce infectious yellows virus (LIYV) RNA 1-encoded P34 is an RNA-binding protein and exhibits perinuclear localization. Virology, 403(1), 67-77. doi:10.1016/j.virol.2010.04.006Weinheimer, I., Boonrod, K., Moser, M., Wassenegger, M., Krczal, G., Butcher, S. J., & Valkonen, J. P. T. (2014). Binding and processing of small dsRNA molecules by the class 1 RNase III protein encoded by sweet potato chlorotic stunt virus. Journal of General Virology, 95(2), 486-495. doi:10.1099/vir.0.058693-0Yeh, H.-H., Tian, T., Rubio, L., Crawford, B., & Falk, B. W. (2000). Asynchronous Accumulation of Lettuce Infectious Yellows Virus RNAs 1 and 2 and Identification of an RNA 1 trans Enhancer of RNA 2 Accumulation. Journal of Virology, 74(13), 5762-5768. doi:10.1128/jvi.74.13.5762-5768.200

Sprouty2 loss‐induced IL6 drives castration‐resistant prostate cancer through scavenger receptor B1

Metastatic castration‐resistant prostate cancer (mCRPC) is a lethal form of treatment‐resistant prostate cancer and poses significant therapeutic challenges. Deregulated receptor tyrosine kinase (RTK) signalling mediated by loss of tumour suppressor Sprouty2 (SPRY2) is associated with treatment resistance. Using pre‐clinical human and murine mCRPC models, we show that SPRY2 deficiency leads to an androgen self‐sufficient form of CRPC. Mechanistically, HER2‐IL6 signalling axis enhances the expression of androgen biosynthetic enzyme HSD3B1 and increases SRB1‐mediated cholesterol uptake in SPRY2‐deficient tumours. Systemically, IL6 elevated the levels of circulating cholesterol by inducing host adipose lipolysis and hepatic cholesterol biosynthesis. SPRY2‐deficient CRPC is dependent on cholesterol bioavailability and SRB1‐mediated tumoral cholesterol uptake for androgen biosynthesis. Importantly, treatment with ITX5061, a clinically safe SRB1 antagonist, decreased treatment resistance. Our results indicate that cholesterol transport blockade may be effective against SPRY2‐deficient CRPC

Sleeping Beauty screen reveals Pparg activation in metastatic prostate cancer

Prostate cancer (CaP) is the most common adult male cancer in the developed world. The paucity of biomarkers to predict prostate tumor biology makes it important to identify key pathways that confer poor prognosis and guide potential targeted therapy. Using a murine forward mutagenesis screen in a Pten-null background, we identified peroxisome proliferator-activated receptor gamma (Pparg), encoding a ligand-activated transcription factor, as a promoter of metastatic CaP through activation of lipid signaling pathways, including up-regulation of lipid synthesis enzymes [fatty acid synthase (FASN), acetyl-CoA carboxylase (ACC), ATP citrate lyase (ACLY)]. Importantly, inhibition of PPARG suppressed tumor growth in vivo, with down-regulation of the lipid synthesis program. We show that elevated levels of PPARG strongly correlate with elevation of FASN in human CaP and that high levels of PPARG/FASN and PI3K/pAKT pathway activation confer a poor prognosis. These data suggest that CaP patients could be stratified in terms of PPARG/FASN and PTEN levels to identify patients with aggressive CaP who may respond favorably to PPARG/FASN inhibition

Analysis of Nkx3.1:Cre-driven Erk5 deletion reveals a profound spinal deformity which is linked to increased osteoclast activity

Extracellular signal-regulated protein kinase 5 (ERK5) has been implicated during development and carcinogenesis. Nkx3.1-mediated Cre expression is a useful strategy to genetically manipulate the mouse prostate. While grossly normal at birth, we observed an unexpected phenotype of spinal protrusion in Nkx3.1:Cre;Erk5fl/fl (Erk5fl/fl) mice by ~6–8 weeks of age. X-ray, histological and micro CT (µCT) analyses showed that 100% of male and female Erk5fl/fl mice had a severely deformed curved thoracic spine, with an associated loss of trabecular bone volume. Although sex-specific differences were observed, histomorphometry measurements revealed that both bone resorption and bone formation parameters were increased in male Erk5fl/fl mice compared to wild type (WT) littermates. Osteopenia occurs where the rate of bone resorption exceeds that of bone formation, so we investigated the role of the osteoclast compartment. We found that treatment of RANKL-stimulated primary bone marrow-derived macrophage (BMDM) cultures with small molecule ERK5 pathway inhibitors increased osteoclast numbers. Furthermore, osteoclast numbers and expression of osteoclast marker genes were increased in parallel with reduced Erk5 expression in cultures generated from Erk5fl/fl mice compared to WT mice. Collectively, these results reveal a novel role for Erk5 during bone maturation and homeostasis in vivo