Lung metastases share common immune features regardless of primary tumor origin

BACKGROUND: Only certain disseminated cells are able to grow in secondary organs to create a metastatic tumor. Under the hypothesis that the immune microenvironment of the host tissue may play an important role in this process, we have categorized metastatic samples based on their immune features. METHODS: Gene expression data of metastatic samples (n=374) from four secondary sites (brain, bone, liver and lung) were used to characterize samples based on their immune and stromal infiltration using gene signatures and cell quantification tools. A clustering analysis was done that separated metastatic samples into three different immune categories: high, medium and low. RESULTS: Significant differences were found between the immune profiles of samples metastasizing in distinct organs. Metastases in lung showed a higher immunogenic score than metastases in brain, liver or bone, regardless of their primary site of origin. Also, they preferentially clustered in the high immune group. Samples in this cluster exhibited a clear inflammatory phenotype, higher levels of immune infiltrate, overexpression of programmed death-ligand 1 (PD-L1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA4) pathways and upregulation of genes predicting clinical response to programmed cell death protein 1 (PD-1) blockade (T-cell inflammatory signature). A decision tree algorithm was used to select CD74 as a biomarker that identify samples belonging to this high-immune subtype of metastases, having specificity of 0.96 and sensitivity of 1. CONCLUSIONS: We have found a group of lung-enriched metastases showing an inflammatory phenotype susceptible to be treated with immunotherapy

In-flight Diagnostics in LISA Pathfinder

LISA PathFinder (LPF) will be flown with the objective to test in space key technologies for LISA. However its sensitivity goals are, for good reason, one order of magnitude less than those which LISA will have to meet, both in drag-free and optical metrology requirements, and in the observation frequency band. While the expected success of LPF will of course be of itself a major step forward to LISA, one might not forget that a further improvement by an order of magnitude in performance will still be needed. Clues for the last leap are to be derived from proper disentanglement of the various sources of noise which contribute to the total noise, as measured in flight during the PathFinder mission. This paper describes the principles, workings and requirements of one of the key tools to serve the above objective: the diagnostics subsystem. This consists in sets of temperature, magnetic field, and particle counter sensors, together with generators of controlled thermal and magnetic perturbations. At least during the commissioning phase, the latter will be used to identify feed-through coefficients between diagnostics sensor readings and associated actual noise contributions. A brief progress report of the current state of development of the diagnostics subsystem will be given as well.Peer Reviewe

In-flight Diagnostics in LISA Pathfinder

LISA PathFinder (LPF) will be flown with the objective to test in space key technologies for LISA. However its sensitivity goals are, for good reason, one order of magnitude less than those which LISA will have to meet, both in drag-free and optical metrology requirements, and in the observation frequency band. While the expected success of LPF will of course be of itself a major step forward to LISA, one might not forget that a further improvement by an order of magnitude in performance will still be needed. Clues for the last leap are to be derived from proper disentanglement of the various sources of noise which contribute to the total noise, as measured in flight during the PathFinder mission. This paper describes the principles, workings and requirements of one of the key tools to serve the above objective: the diagnostics subsystem. This consists in sets of temperature, magnetic field, and particle counter sensors, together with generators of controlled thermal and magnetic perturbations. At least during the commissioning phase, the latter will be used to identify feed-through coefficients between diagnostics sensor readings and associated actual noise contributions. A brief progress report of the current state of development of the diagnostics subsystem will be given as well.Peer Reviewe

Fitness Ranking of Individual Mutants Drives Patterns of Epistatic Interactions in HIV-1

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Rainwater harvesting systems reduce detergent use

Unidad de excelencia María de Maeztu MdM-2015-0552Purpose: Due to population growth, urban water demand is expected to increase significantly, as well as the environmental and economic costs required to supply it. Rainwater harvesting (RWH) systems can play a key role in helping cities meet part of their water demand as an alternative to conventional water abstraction and treatment. This paper presents an environmental and economic analysis of RWH systems providing households with water for laundry purposes in a life cycle thinking perspective. Conclusions: LCA and LCC present better results for high-density scenarios. Overall, avoided environmental and economic impacts from detergent reduction clearly surpass environmental impacts (in all categories except terrestrial acidification) and economic cost of the RWHsystem in most cases (except two scenarios). Another important finding is that 80%of the savings are achieved by minimizing detergent and fabric softener by using soft rainwater; and the remaining 20% comes from replacing the use of tap water

TLR9 agonists induced cell death in Burkitt's lymphoma cells is variable and influenced by TLR9 polymorphism

Toll-like receptor 9 (TLR9) triggering is a promising novel strategy to combat cancer as it induces innate and adaptive immunity responses. B-cell lymphoma is unique in this context as tumor cells express TLR9 and may harbor latent Epstein-Barr virus (EBV), a gamma-herpesvirus with remarkable oncogenic potential when latent. Latent EBV may be promoted by TLR9 triggering via suppression of lytic EBV. Here, we elaborated an initial assessment of the impact of TLR9 triggering on EBV-positive and EBV-negative B-cell lymphoma using Burkitt's lymphoma (BL) cell lines as an in vitro model. We show that, independent of the presence of EBV, the TLR9 ligand oligodeoxynucleotide (ODN) CpG-2006 may or may not induce caspase-dependent cell death in BL cells. Moreover, ODN CpG-2006-induced cell death responses of BL cells were associated with TLR9 single-nucleotide polymorphisms (SNPs) rs5743836 or rs352140, which we detected in primary BL tumors and in peripheral blood from healthy individuals at similar frequencies. Thus, our findings suggest that the effect of TLR9 agonists on BL cells should be tested in vitro before installment of therapy and TLR9 SNPs in BL patients should be determined as potential biological markers for the therapeutic response to treatment targeting innate immunity

Alternative initiation and splicing in dicer gene expression in human breast cells

INTRODUCTION: Dicer is a ribonuclease that mediates RNA interference both at the transcriptional and the post-transcriptional levels. Human dicer gene expression is regulated in different tissues. Dicer is responsible for the synthesis of microRNAs and short temporal (st)RNAs that regulate the expression of many genes. Thus, understanding the control of the expression of the dicer gene is essential for the appreciation of double-stranded (ds)RNA-mediated pathways of gene expression. Human dicer mRNA has many upstream open reading frames (uORFs) at the 5'-leader sequences (the nucleotide sequence between the 5'-end and the start codon of the major ORF), and we studied whether these elements at the 5'-leader sequences regulate the expression of the dicer gene. METHOD: We determined the 5'-leader sequences of the dicer mRNAs in human breast cells by 5'-RACE and S1-nuclease protection analysis. We have analyzed the functions of the 5'-leader variants by reporter gene expression in vitro and in vivo. RESULTS: We found that the dicer transcripts in human breast cells vary in the sequence of their 5'-leader sequences, and that alternative promoter selection along with alternative splicing of the 5'-terminal exons apparently generate these variations. The breast cell has at least two predominant forms of dicer mRNAs, one of which has an additional 110 nucleotides at the 5'-end. Sequence comparison revealed that the first 80 nucleotides of these mRNA isoforms are encoded by a new exon located approximately 16 kb upstream of the reported start site. There are 30 extra nucleotides added to the previously reported exon 1. The human breast cells studied predominantly express two 5'-leader variants of dicer mRNAs, one with the exons 2 and 3 (long form) and the other without them (short form). By reporter gene expression analysis we found that the exon 2 and 3 sequences at the 5'-leader sequences are greatly inhibitory for the translation of the mRNA into protein. CONCLUSION: Dicer gene expression in human breast cells is regulated by alternative promoter selection to alter the length and composition of the 5'-leader sequence of its mRNA. Furthermore, alternative splicing of its exon 2 and 3 sequences of their pre-mRNA creates a more translationally competent mRNA in these cells

Efficient derivation of NPCs, spinal motor neurons and midbrain dopaminergic neurons from hESCs at 3% oxygen

This protocol has been designed to generate neural precursor cells (NPCs) from human embryonic stem cells (hESCs) using a physiological oxygen (O(2)) level of 3% and chemically defined conditions. The first stage involves suspension culture of hESC colonies at 3% O(2), where they acquire a neuroepithelial identity over two weeks. This timescale is comparable to that at 20% O(2), but survival is enhanced. Sequential application of retinoic acid (RA) and purmorphamine (PM), from day 14 to 28, directs differentiation towards spinal motor neurons. Alternatively, addition of FGF-8 and PM generates midbrain dopaminergic neurons. OLIG2 induction in motor neuron precursors is 2-fold greater than at 20% O(2), whereas EN1 is 5-fold enhanced. 3% NPCs can be differentiated into all three neural lineages, and such cultures can be maintained long-term in the absence of neurotrophins. The ability to generate defined cell types at 3% O(2) should represent a significant advance for in vitro disease modelling and potentially cell-based therapies