Molecular targets for the protodynamic action of cis-urocanic acid in human bladder carcinoma cells

<p>Abstract</p> <p>Background</p> <p>cis-urocanic acid (cis-UCA) is an endogenous amino acid metabolite capable of transporting protons from the mildly acidic extracellular medium into the cell cytosol. The resulting intracellular acidification suppresses many cellular activities. The current study was aimed at characterizing the molecular mechanisms underlying cis-UCA-mediated cytotoxicity in cultured cancer cells.</p> <p>Methods</p> <p>5367 bladder carcinoma cells were left untreated or treated with cis-UCA. Cell death was assessed by measuring caspase-3 activity, mitochondrial membrane polarization, formation and release of cytoplasmic histone-associated DNA fragments, and cellular permeabilization. Cell viability and metabolic activity were monitored by colorimetric assays. Nuclear labelling was used to quantify the effects of cis-UCA on cell cycle. The activity of the ERK and JNK signalling pathways was studied by immunoblotting with specific antibodies. Phosphatase activity in cis-UCA-treated cells was determined by assay kits measuring absorbance resulting from the dephosphorylation of an artificial substrate. All statistical analyses were performed using the two-way Student's t-test (p < 0.05).</p> <p>Results</p> <p>Here we report that treatment of the 5637 human bladder carcinoma cells with 2% cis-UCA induces both apoptotic and necrotic cell death. In addition, metabolic activity of the 5637 cells is rapidly impaired, and the cells arrest in cell cycle in response to cis-UCA. Importantly, we show that cis-UCA promotes the ERK and JNK signalling pathways by efficiently inhibiting the activity of serine/threonine and tyrosine phosphatases.</p> <p>Conclusions</p> <p>Our studies elucidate how cis-UCA modulates several cellular processes, thereby inhibiting the proliferation and survival of bladder carcinoma cells. These anti-cancer effects make cis-UCA a potential candidate for the treatment of non-muscle invasive bladder carcinoma.</p

Induction of humoral immune response to multiple recombinant Rhipicephalus appendiculatus antigens and their effect on tick feeding success and pathogen transmission

BACKGROUND: Rhipicephalus appendiculatus is the primary vector of Theileria parva, the etiological agent of East Coast fever (ECF), a devastating disease of cattle in sub-Saharan Africa. We hypothesized that a vaccine targeting tick proteins that are involved in attachment and feeding might affect feeding success and possibly reduce tick-borne transmission of T. parva. Here we report the evaluation of a multivalent vaccine cocktail of tick antigens for their ability to reduce R. appendiculatus feeding success and possibly reduce tick-transmission of T. parva in a natural host-tick-parasite challenge model. METHODS: Cattle were inoculated with a multivalent antigen cocktail containing recombinant tick protective antigen subolesin as well as two additional R. appendiculatus saliva antigens: the cement protein TRP64, and three different histamine binding proteins. The cocktail also contained the T. parva sporozoite antigen p67C. The effect of vaccination on the feeding success of nymphal and adult R. appendiculatus ticks was evaluated together with the effect on transmission of T. parva using a tick challenge model. RESULTS: To our knowledge, this is the first evaluation of the anti-tick effects of these antigens in the natural host-tick-parasite combination. In spite of evidence of strong immune responses to all of the antigens in the cocktail, vaccination with this combination of tick and parasite antigens did not appear to effect tick feeding success or reduce transmission of T. parva. CONCLUSION: The results of this study highlight the importance of early evaluation of anti-tick vaccine candidates in biologically relevant challenge systems using the natural tick-host-parasite combination

Rethinking Radical Democracy

Over the course of three decades, vocabularies of radical democracy have pressed their stamp on democratic thought. Trading on the intuition that there is more to democracy than elections, they have generated critical insights into the important role that practices of pluralisation and critique play in bettering institutional politics. As a result, few would today deny the radical democratic contribution to democratic thought. What many might question, however, is its continuing traction. The paper probes this question, focusing on the nuanced place of democracy in contemporary radical work. It grapples with the difficulties that this poses for radical democrats and it suggests that a way of overcoming these difficulties – that threaten to undermine the coherence of radical democracy – is to rethink and reconstruct the distinctiveness of its vocabulary. The paper attempts such a reconstruction. It develops the idea of the promissory rule of the many and it discusses the ways it rejuvenates broader democratic thought

Climate, human behaviour or environment: individual-based modelling of Campylobacter seasonality and strategies to reduce disease burden

Acknowledgements: We thank colleagues within the Modelling, Evidence and Policy Research Group for useful feedback on this manuscript. Competing interests: The authors declare that they have no competing interests. Availability of data and materials: The R code used in this research is available at https://gitlab.com/rasanderson/campylobacter-microsimulation; it is platform independent, R version 3.3.0 and above. Funding: This research was funded by Medical Research Council Grant, Natural Environment Research Council, Economic and Social Research Council, Biotechnology and Biological Sciences Research Council, and the Food Standards Agency through the Environmental and Social Ecology of Human Infectious Diseases Initiative (Sources, seasonality, transmission and control: Campylobacter and human behaviour in a changing environment (ENIGMA); Grant Reference G1100799-1). PRH, SJO’B, and IRL are funded in part by the NIHR Health Protection Research Unit in Gastrointestinal Infection, at the University of Liverpool. PRH and IRL are also funded in part by the NIHR Health Protection Research Unit in Emergency Preparedness and Response, at King’s College London. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, the Department of Health or Public Health England.Peer reviewedPublisher PD