Cellular immunotherapy of murine neuroblastoma.

Neuroblastoma (NB) is the most common extracranial paediatric tumour, and patients with disseminated disease have a poor long-term prognosis. Due to the significant mortality rate, alternative treatments to conventional therapies are continually sought. Using the A/J mouse model it has previously been demonstrated that a cellular vaccine of the syngeneic Neuro-2a NB cell line modified to express IL-2 and IL-12 abrogated the tumourigenicity of Neuro-2a cells, and mediated regression of established tumours. However, establishing cultures of primary NB cells can be problematic, making this approach difficult to implement clinically. This thesis describes the development of an alternative cellular vaccine to treat murine NB using a synthetic vector. Firstly, transfection of primary dendritic cells was optimised. Dendritic cells are potent antigen presenting cells and studies have shown them to generate anti-cancer responses. Optimal transfection levels of 5% were obtained but antigen presentation by these cells was limited. Therefore, an alternative approach was developed using fibroblasts engineered to express IL-2 and IL-12. Cytokine-expressing fibroblasts could be used in place of transfected tumour cells to provide sustained, high-level cytokine expression in the tumour locale. Transfection of syngeneic and allogeneic murine fibroblasts was optimised in vitro to produce therapeutic levels of IL-2 and IL-12. Cytokine-transfected fibroblasts were compared with cytokine-transfected Neuro-2a cells to prevent engraftment of wild-type Neuro-2a cells in vivo. The allogeneic cells prevented tumour engraftment in a nonspecific, cytokine-independent manner. Syngeneic fibroblasts expressing IL-2 and IL-12 inhibited tumour engraftment as effectively as cytokine expressing Neuro-2a cells, and this rejection was cytokine-dependent. The cytokine-transfected syngeneic fibroblasts induced protective immunity against rechallenge with wild-type Neuro-2a cells as effectively as cytokine-transfected-Neuro-2a cells. Intratumoural vaccination of cytokine-transfected syngeneic fibroblasts also demonstrated therapeutic efficacy against Neuro-2a-derived established tumours. Splenocytes from vaccinated mice demonstrated increased IL-2 and IFN-y expression and cytotoxicity compared with controls when co-cultured with wild-type Neuro-2a cells in vitro. Vaccinated tumours showed decreased vascularity and increased infiltration of CD45+ cells compared with controls. Therefore, cytokine-transfected syngeneic fibroblasts are a viable potential alternative vaccine for the treatment of minimal residual NB

Packaging signals in single-stranded RNA viruses: nature’s alternative to a purely electrostatic assembly mechanism

The formation of a protective protein container is an essential step in the life-cycle of most viruses. In the case of single-stranded (ss)RNA viruses, this step occurs in parallel with genome packaging in a co-assembly process. Previously, it had been thought that this process can be explained entirely by electrostatics. Inspired by recent single-molecule fluorescence experiments that recapitulate the RNA packaging specificity seen in vivo for two model viruses, we present an alternative theory, which recognizes the important cooperative roles played by RNA–coat protein interactions, at sites we have termed packaging signals. The hypothesis is that multiple copies of packaging signals, repeated according to capsid symmetry, aid formation of the required capsid protein conformers at defined positions, resulting in significantly enhanced assembly efficiency. The precise mechanistic roles of packaging signal interactions may vary between viruses, as we have demonstrated for MS2 and STNV. We quantify the impact of packaging signals on capsid assembly efficiency using a dodecahedral model system, showing that heterogeneous affinity distributions of packaging signals for capsid protein out-compete those of homogeneous affinities. These insights pave the way to a new anti-viral therapy, reducing capsid assembly efficiency by targeting of the vital roles of the packaging signals, and opens up new avenues for the efficient construction of protein nanocontainers in bionanotechnology

Quantitation and localization of ENaC subunit expression in fetal, newborn, and adult mouse lung

The newborn lung is cleared of fetal liquid by active Na+ transport. The heterotrimeric (α, β, γ) epithelial Na+ channel, ENaC, mediates this process. To understand the role of individual ENaC subunits in Na+ transport during development, we quantified murine ENaC (mENaC) subunit messenger RNA (mRNA) expression levels of fetal, neonatal, and adult mouse lung by Northern blot analysis and studied regional expression by in situ hybridization. αmENaC and γmENaC mRNA expression increased sharply in late fetal gestation and reached near-adult levels by Day 1 of postnatal life. βmENaC expression increased more gradually through late fetal and early postnatal life and increased progressively until adulthood. In situ hybridization studies showed similar localization patterns of αmENaC and γmENaC subunit expression in fetal and postnatal lung. γmENaC and αmENaC subunits were initially localized to fetal lung bud tubules and by late gestation both subunits were expressed in all regions (acinar and bronchiolar) of the distal lung epithelium. βmENaC was detected from 16 d gestation onward and was expressed most intensely in small airways. There was little expression of βmENaC in the alveolar region. In postnatal lung all three subunits were expressed intensely in small airways. In adult lung, αmENaC and γmENaC were expressed in a pattern consistent with an alveolar type II (ATII) cell distribution. The timing of quantitative changes in mENaC subunit expression is consistent with a role of Na+ transport in liquid clearance of the perinatal lung. Intense expression of mENaC subunits in medium and small airway epithelium and in ATII cells suggests that these regions are a primary location for liquid absorption in the perinatal and post-natal murine lung

Height and risk of death among men and women: aetiological implications of associations with cardiorespiratory disease and cancer mortality

OBJECTIVES: Height is inversely associated with cardiovascular disease mortality risk and has shown variable associations with cancer incidence and mortality. The interpretation of findings from previous studies has been constrained by data limitations. Associations between height and specific causes of death were investigated in a large general population cohort of men and women from the West of Scotland. DESIGN: Prospective observational study. SETTING: Renfrew and Paisley, in the West of Scotland. SUBJECTS: 7052 men and 8354 women aged 45-64 were recruited into a study in Renfrew and Paisley, in the West of Scotland, between 1972 and 1976. Detailed assessments of cardiovascular disease risk factors, morbidity and socioeconomic circumstances were made at baseline. MAIN OUTCOME MEASURES: Deaths during 20 years of follow up classified into specific causes. RESULTS: Over the follow up period 3347 men and 2638 women died. Height is inversely associated with all cause, coronary heart disease, stroke, and respiratory disease mortality among men and women. Adjustment for socioeconomic position and cardiovascular risk factors had little influence on these associations. Height is strongly associated with forced expiratory volume in one second (FEV1) and adjustment for FEV1 considerably attenuated the association between height and cardiorespiratory mortality. Smoking related cancer mortality is not associated with height. The risk of deaths from cancer unrelated to smoking tended to increase with height, particularly for haematopoietic, colorectal and prostate cancers. Stomach cancer mortality was inversely associated with height. Adjustment for socioeconomic position had little influence on these associations. CONCLUSION: Height serves partly as an indicator of socioeconomic circumstances and nutritional status in childhood and this may underlie the inverse associations between height and adulthood cardiorespiratory mortality. Much of the association between height and cardiorespiratory mortality was accounted for by lung function, which is also partly determined by exposures acting in childhood. The inverse association between height and stomach cancer mortality probably reflects Helicobacter pylori infection in childhood resulting inor being associated withshorter height. The positive associations between height and several cancers unrelated to smoking could reflect the influence of calorie intake during childhood on the risk of these cancers

From Pre‐Roman Bailo To Roman Baelo: Long‐Term Landscape Dynamics In The Straits Of Gibraltar

The Straits of Gibraltar have been historically an important maritime axis of connection between the Mediterranean and Atlantic areas of the Iberian Peninsula. For this reason, most of the archaeological research has focused on the coastal settlements, but its broader archaeological landscape remains mostly unknown. In this paper, we present recent intensive surveys in which a wide range of sites was detected, dating from the eighth century BC to the fourteenth AD. The ancient landscape is thus reconstructed over a long‐term perspective. Prior to the Roman expansion, the earlier Bailo‐La Silla del Papa was an urban central place that supported a dense network of subordinate settlements. Later on, the central settlement was transported from inland to the coastal town of Baelo Claudia, but the territorial structure remained based on a similar pattern.This study has been carried out as part of the Franco-German (Agence Nationale de la Recherche Scientifique and Deutsche Forschungsgemeinschaft) project – ARCHEOSTRAITS. Espaces protohistoriques du détroit de Gibraltar: les territoires de la Silla del Papa et de Los Castillejos de Alcorrín (IXe – Ier siècle av. J.-C.) – led by D.Marzoli and P.Moret. In this article we have presented the analysis of the Atlantic sector, drawing on a study included in a Junta de Andalucía General Research Project led by P. Moret – La Silla del Papa (Tarifa, Cádiz): oppidum, necropolis and territory (2014-2019)

The Geological Record of Ocean Acidification

Ocean acidification may have severe consequences for marine ecosystems; however, assessing its future impact is difficult because laboratory experiments and field observations are limited by their reduced ecologic complexity and sample period, respectively. In contrast, the geological record contains long-term evidence for a variety of global environmental perturbations, including ocean acidification plus their associated biotic responses. We review events exhibiting evidence for elevated atmospheric CO2, global warming, and ocean acidification over the past ~300 million years of Earth's history, some with contemporaneous extinction or evolutionary turnover among marine calcifiers. Although similarities exist, no past event perfectly parallels future projections in terms of disrupting the balance of ocean carbonate chemistry—a consequence of the unprecedented rapidity of CO2 release currently taking place

Detector Description and Performance for the First Coincidence Observations between LIGO and GEO

For 17 days in August and September 2002, the LIGO and GEO interferometer gravitational wave detectors were operated in coincidence to produce their first data for scientific analysis. Although the detectors were still far from their design sensitivity levels, the data can be used to place better upper limits on the flux of gravitational waves incident on the earth than previous direct measurements. This paper describes the instruments and the data in some detail, as a companion to analysis papers based on the first data.Comment: 41 pages, 9 figures 17 Sept 03: author list amended, minor editorial change

Analysis of LIGO data for gravitational waves from binary neutron stars

We report on a search for gravitational waves from coalescing compact binary systems in the Milky Way and the Magellanic Clouds. The analysis uses data taken by two of the three LIGO interferometers during the first LIGO science run and illustrates a method of setting upper limits on inspiral event rates using interferometer data. The analysis pipeline is described with particular attention to data selection and coincidence between the two interferometers. We establish an observational upper limit of $\mathcal{R}<$1.7 \times 10^{2}$per year per Milky Way Equivalent Galaxy (MWEG), with 90coalescence rate of binary systems in which each component has a mass in the range 1--3$M_\odot$.Comment: 17 pages, 9 figure

Isotopic signatures of methane emissions from tropical fires, agriculture and wetlands: the MOYA and ZWAMPS flights

We report methane isotopologue data from aircraft and ground measurements in Africa and South America. Aircraft campaigns sampled strong methane fluxes over tropical papyrus wetlands in the Nile, Congo and Zambezi basins, herbaceous wetlands in Bolivian southern Amazonia, and over fires in African woodland, cropland and savannah grassland. Measured methane δ13CCH4 isotopic signatures were in the range −55 to −49‰ for emissions from equatorial Nile wetlands and agricultural areas, but widely −60 ± 1‰ from Upper Congo and Zambezi wetlands. Very similar δ13CCH4 signatures were measured over the Amazonian wetlands of NE Bolivia (around −59‰) and the overall δ13CCH4 signature from outer tropical wetlands in the southern Upper Congo and Upper Amazon drainage plotted together was −59 ± 2‰. These results were more negative than expected. For African cattle, δ13CCH4 values were around −60 to −50‰. Isotopic ratios in methane emitted by tropical fires depended on the C3 : C4 ratio of the biomass fuel. In smoke from tropical C3 dry forest fires in Senegal, δ13CCH4 values were around −28‰. By contrast, African C4 tropical grass fire δ13CCH4 values were −16 to −12‰. Methane from urban landfills in Zambia and Zimbabwe, which have frequent waste fires, had δ13CCH4 around −37 to −36‰. These new isotopic values help improve isotopic constraints on global methane budget models because atmospheric δ13CCH4 values predicted by global atmospheric models are highly sensitive to the δ13CCH4 isotopic signatures applied to tropical wetland emissions. Field and aircraft campaigns also observed widespread regional smoke pollution over Africa, in both the wet and dry seasons, and large urban pollution plumes. The work highlights the need to understand tropical greenhouse gas emissions in order to meet the goals of the UNFCCC Paris Agreement, and to help reduce air pollution over wide regions of Africa

Use of SMS texts for facilitating access to online alcohol interventions: a feasibility study

A41 Use of SMS texts for facilitating access to online alcohol interventions: a feasibility study In: Addiction Science & Clinical Practice 2017, 12(Suppl 1): A4