Targeted knock-down of miR21 primary transcripts using snoMEN vectors induces apoptosis in human cancer cell lines

We have previously reported an antisense technology, 'snoMEN vectors', for targeted knock-down of protein coding mRNAs using human snoRNAs manipulated to contain short regions of sequence complementarity with the mRNA target. Here we characterise the use of snoMEN vectors to target the knock-down of micro RNA primary transcripts. We document the specific knock-down of miR21 in HeLa cells using plasmid vectors expressing miR21-targeted snoMEN RNAs and show this induces apoptosis. Knock-down is dependent on the presence of complementary sequences in the snoMEN vector and the induction of apoptosis can be suppressed by over-expression of miR21. Furthermore, we have also developed lentiviral vectors for delivery of snoMEN RNAs and show this increases the efficiency of vector transduction in many human cell lines that are difficult to transfect with plasmid vectors. Transduction of lentiviral vectors expressing snoMEN targeted to pri-miR21 induces apoptosis in human lung adenocarcinoma cells, which express high levels of miR21, but not in human primary cells. We show that snoMEN-mediated suppression of miRNA expression is prevented by siRNA knock-down of Ago2, but not by knock-down of Ago1 or Upf1. snoMEN RNAs colocalise with Ago2 in cell nuclei and nucleoli and can be co-immunoprecipitated from nuclear extracts by antibodies specific for Ago2

Mercaptobenzothiazole or the mercapto-mix: which should be in the standard series?

Mercaptobenzothiazole (MBT) compounds are well known contact allergens. To detect rubber allergic patients we use both MBT (2% in petrolatum) and a mercapto-mix with 4 constituents of 0.5% each in our standard series. In this article the EECDRG presents data of in total 32,475 consecutive tested patients attending the respective contact dermatitis clinics from 11 centres in Europe to determine if the mix and MBT detected the same allergic patients. We found 327 patients positive to the mix or MBT, or to both. 261 were positive to the mix and 254 to MBT. MBT was negative in 73 patients who were positive to the mix. If the mix had not been in the standard series, on average 22% of patients allergic to a mercapto-compound would have been missed, for MBT this would have been on average 20%. All clinics would have missed a significant number of positive reactions if both compounds had not been tested. We conclude, that both the mercapto mix and MBT are required in the standard series

Volatility forecasting in the Chinese commodity futures market with intraday data

Given the unique institutional regulations in the Chinese commodity futures market as well as the characteristics of the data it generates, we utilize contracts with three months to delivery, the most liquid contract series, to systematically explore volatility forecasting for aluminum, copper, fuel oil, and sugar at the daily and three intraday sampling frequencies. We adopt popular volatility models in the literature and assess the forecasts obtained via these models against alternative proxies for the true volatility. Our results suggest that the long memory property is an essential feature in the commodity futures volatility dynamics and that the ARFIMA model consistently produces the best forecasts or forecasts not inferior to the best in statistical terms

Membranes by the Numbers

Many of the most important processes in cells take place on and across membranes. With the rise of an impressive array of powerful quantitative methods for characterizing these membranes, it is an opportune time to reflect on the structure and function of membranes from the point of view of biological numeracy. To that end, in this article, I review the quantitative parameters that characterize the mechanical, electrical and transport properties of membranes and carry out a number of corresponding order of magnitude estimates that help us understand the values of those parameters.Comment: 27 pages, 12 figure

Contemporary carbon fluxes do not reflect the long-term carbon balance for an Atlantic blanket bog

Peatlands are one of the largest terrestrial stores of carbon. Carbon exchange in peatlands is often assessed solely by measurement of contemporary fluxes; however, these fluxes frequently indicate a much stronger sink strength than that measured by the rate of C accumulation in the peat profile over longer timescales. Here we compare profile-based measurements of C accumulation with the published net ecosystem C balance for the largest peatland area in Britain, the Flow Country of northern Scotland. We estimate the long-term rate of C accumulation to be 15.4 g C m−2 yr−1 for a site where a recent eddy covariance study has suggested contemporary C uptake more than six times greater (99.37 g C m−2 yr−1). Our estimate is supported by two further long-term C accumulation records from nearby sites which give comparable results. We demonstrate that a strong contemporary C sink strength may not equate to a strong long-term sink and explore reasons for this disparity. We recommend that contemporary C sequestration should be viewed in the context of the long-term ecological drivers, such as fires, ecohydrological feedbacks and the changing quality of litter inputs

Vortices in polariton OPO superfluids

This chapter reviews the occurrence of quantised vortices in polariton fluids, primarily when polaritons are driven in the optical parametric oscillator (OPO) regime. We first review the OPO physics, together with both its analytical and numerical modelling, the latter being necessary for the description of finite size systems. Pattern formation is typical in systems driven away from equilibrium. Similarly, we find that uniform OPO solutions can be unstable to the spontaneous formation of quantised vortices. However, metastable vortices can only be injected externally into an otherwise stable symmetric state, and their persistence is due to the OPO superfluid properties. We discuss how the currents charactering an OPO play a crucial role in the occurrence and dynamics of both metastable and spontaneous vortices.Comment: 40 pages, 16 figure

Erosion characteristics and horizontal variability for small erosion depths in the Sacramento-San Joaquin River Delta, California, USA

Erodibility of cohesive sediment in the Sacramento-San Joaquin River Delta (Delta) was investigated with an erosion microcosm. Erosion depths in the Delta and in the microcosm were estimated to be about one floc diameter over a range of shear stresses and times comparable to half of a typical tidal cycle. Using the conventional assumption of horizontally homogeneous bed sediment, data from 27 of 34 microcosm experiments indicate that the erosion rate coefficient increased as eroded mass increased, contrary to theory. We believe that small erosion depths, erosion rate coefficient deviation from theory, and visual observation of horizontally varying biota and texture at the sediment surface indicate that erosion cannot solely be a function of depth but must also vary horizontally. We test this hypothesis by developing a simple numerical model that includes horizontal heterogeneity, use it to develop an artificial time series of suspended-sediment concentration (SSC) in an erosion microcosm, then analyze that time series assuming horizontal homogeneity. A shear vane was used to estimate that the horizontal standard deviation of critical shear stress was about 30% of the mean value at a site in the Delta. The numerical model of the erosion microcosm included a normal distribution of initial critical shear stress, a linear increase in critical shear stress with eroded mass, an exponential decrease of erosion rate coefficient with eroded mass, and a stepped increase in applied shear stress. The maximum SSC for each step increased gradually, thus confounding identification of a single well-defined critical shear stress as encountered with the empirical data. Analysis of the artificial SSC time series with the assumption of a homogeneous bed reproduced the original profile of critical shear stress, but the erosion rate coefficient increased with eroded mass, similar to the empirical data. Thus, the numerical experiment confirms the small-depth erosion hypothesis. A linear model of critical shear stress and eroded mass is proposed to simulate small-depth erosion, assuming that the applied and critical shear stresses quickly reach equilibrium

Mouse Protocadherin-1 gene expression is regulated by cigarette smoke exposure in vivo

Protocadherin-1 (PCDH1) is a novel susceptibility gene for airway hyperresponsiveness, first identified in families exposed to cigarette smoke and is expressed in bronchial epithelial cells. Here, we asked how mouse Pcdh1 expression is regulated in lung structural cells in vivo under physiological conditions, and in both short-term cigarette smoke exposure models characterized by airway inflammation and hyperresponsiveness and chronic cigarette smoke exposure models. Pcdh1 gene-structure was investigated by Rapid Amplification of cDNA Ends. Pcdh1 mRNA and protein expression was investigated by qRT-PCR, western blotting using isoform-specific antibodies. We observed 87% conservation of the Pcdh1 nucleotide sequence, and 96% conservation of the Pcdh1 protein sequence between men and mice. We identified a novel Pcdh1 isoform encoding only the intracellular signalling motifs. Cigarette smoke exposure for 4 consecutive days markedly reduced Pcdh1 mRNA expression in lung tissue (3 to 4-fold), while neutrophilia and airway hyperresponsiveness was induced. Moreover, Pcdh1 mRNA expression in lung tissue was reduced already 6 hours after an acute cigarette-smoke exposure in mice. Chronic exposure to cigarette smoke induced loss of Pcdh1 protein in lung tissue after 2 months, while Pcdh1 protein levels were no longer reduced after 9 months of cigarette smoke exposure. We conclude that Pcdh1 is highly homologous to human PCDH1, encodes two transmembrane proteins and one intracellular protein, and is regulated by cigarette smoke exposure in vivo