Laboratory tests to study the influence of rock stress confinement on the performances of TBM discs in tunnels

To clarify some aspects of rock destruction with a disc acting on a high confined tunnel face, a series of tests were carried out to examine fracture mechanisms under an indenter that simulates the tunnel boring machine (TBM) tool action, in the presence of an adjacent groove, when a state of stress (lateral confinement) is imposed on a rock sample. These tests proved the importance of carefully establishing the optimal distance of grooves produced by discs acting on a confined surface, and the value (as a mere order of magnitude) of the increase of the thrust to produce the initiation of chip formation, as long as the confinement pressure becomes greater. © University of Science and Technology Beijing and Springer-Verlag Berlin Heidelberg 2011.

Italy's got talent. How Italian leading companies in confectionery industry overcame the crisis

The crisis that spread worldwide since 2007 started from the financial sector and ended to affect also real economy. This process has attracted the attention of many scholars seeking to study its causes and impacts. Notwithstanding many works on this topic, the impact of the crisis on specific industries is still rather unexplored. The present work seeks to address this issue by analyzing the confectionery industry, with particular emphasis for Italian market leaders, Ferrero S.p.A. and Perfetti Van Melle S.r.l.. The aim of the study is to assess if they have been successful in tackling the crisis, keeping a satisfactory level of profitability associated to a good financial health notwithstanding ongoing difficulties. Moreover, we seek to analyze the strategies the companies employed to survive the crisis. The concern of the paper is both quantitative and qualitative. Thus, we calculated a complete set of indicators using a specific methodology for financial statement analysis which has been conceived especially for studying Italian firms; these data have been integrated with other information retrieved from the annual reports of the companies (especially the notes to the accounts and the directors’ report). The analysis highlights that both the firms benefit from a good financial health, with Perfetti Van Melle presenting a large amount of liquidity. On the contrary, liquidity should be the main concern of Ferrero because of an excessive reliance on current liabilities. Both the firms have a good level of profitability, even if Perfetti Van Melle’s one is decreasing. The key-strategies for the success of these firms are ongoing investments in state-of-the-art plant and machinery, an increasing use of equity as the main source of funding, along with huge investments in research and advertising.

Identification of a BRCA1-mRNA Splicing Complex Required for Efficient DNA Repair and Maintenance of Genomic Stability

Mutations within BRCA1 predispose carriers to a high risk of breast and ovarian cancers. BRCA1 functions to maintain genomic stability through the assembly of multiple protein complexes involved in DNA repair, cell-cycle arrest, and transcriptional regulation. Here, we report the identification of a DNA damage-induced BRCA1 protein complex containing BCLAF1 and other key components of the mRNA-splicing machinery. In response to DNA damage, this complex regulates pre-mRNA splicing of a number of genes involved in DNA damage signaling and repair, thereby promoting the stability of these transcripts/proteins. Further, we show that abrogation of this complex results in sensitivity to DNA damage, defective DNA repair, and genomic instability. Interestingly, mutations in a number of proteins found within this complex have been identified in numerous cancer types. These data suggest that regulation of splicing by the BRCA1-mRNA splicing complex plays an important role in the cellular response to DNA damage.

Low-Power Process-Variation Tolerant Arithmetic Units Using Input-Based Elastic Clocking

In this paper we propose a design methodology for low-power high-performance, process-variation tolerant architecture for arithmetic units. The novelty of our approach lies in the fact that possible delay failures due to process variations and/or voltage scaling are predicted in advance and addressed by employing an elastic clocking technique. The prediction mechanism exploits the dependence of delay of arithmetic units upon input data patterns and identifies specific inputs that activate the critical path. Under iso-yield conditions, the proposed design operates at a lower scaled down Vdd without any performance degradation, while it ensures a superlative yield under a design style employing nominal supply and transistor threshold voltage. Simulation results show power savings of upto 29%, energy per computation savings of upto 25.5% and yield enhancement of upto 11.1% compared to the conventional adders and multipliers implemented in the 70nm BPTM technology. We incorporated the proposed modules in the execution unit of a five stage DLX pipeline to measure performance using SPEC2000 benchmarks [9]. Maximum area and throughput penalty obtained were 10% and 3% respectively.

Significance Driven Computation: A Voltage-Scalable, Variation-Aware, Quality-Tuning Motion Estimator

In this paper we present a design methodology for algorithm/architecture co-design of a voltage-scalable, process variation aware motion estimator based on significance driven computation. The fundamental premise of our approach lies in the fact that all computations are not equally significant in shaping the output response of video systems. We use a statistical technique to intelligently identify these significant/not-so-significant computations at the algorithmic level and subsequently change the underlying architecture such that the significant computations are computed in an error free manner under voltage over-scaling. Furthermore, our design includes an adaptive quality compensation (AQC) block which "tunes" the algorithm and architecture depending on the magnitude of voltage over-scaling and severity of process variations. Simulation results show average power savings of similar to 33% for the proposed architecture when compared to conventional implementation in the 90 nm CMOS technology. The maximum output quality loss in terms of Peak Signal to Noise Ratio (PSNR) was similar to 1 dB without incurring any throughput penalty.

Significance Driven Computation on Next-Generation Unreliable Platforms

In this paper, we propose a design paradigm for energy efficient and variation-aware operation of next-generation multicore heterogeneous platforms. The main idea behind the proposed approach lies on the observation that not all operations are equally important in shaping the output quality of various applications and of the overall system. Based on such an observation, we suggest that all levels of the software design stack, including the programming model, compiler, operating system (OS) and run-time system should identify the critical tasks and ensure correct operation of such tasks by assigning them to dynamically adjusted reliable cores/units. Specifically, based on error rates and operating conditions identified by a sense-and-adapt (SeA) unit, the OS selects and sets the right mode of operation of the overall system. The run-time system identifies the critical/less-critical tasks based on special directives and schedules them to the appropriate units that are dynamically adjusted for highly-accurate/approximate operation by tuning their voltage/frequency. Units that execute less significant operations can operate at voltages less than what is required for correct operation and consume less power, if required, since such tasks do not need to be always exact as opposed to the critical ones. Such scheme can lead to energy efficient and reliable operation, while reducing the design cost and overheads of conventional circuit/micro-architecture level techniques.

USP17 is required for clathrin mediated endocytosis of epidermal growth factor receptor

Previously we have shown that expression of the deubiquitinating enzyme USP17 is required for cell proliferation and motility. More recently we reported that USP17 deubiquitinates RCE1 isoform 2 and thus regulates the processing of 'CaaX' motif proteins. Here we now show that USP17 expression is induced by epidermal growth factor and that USP17 expression is required for clathrin mediated endocytosis of epidermal growth factor receptor. In addition, we show that USP17 is required for the endocytosis of transferrin, an archetypal substrate for clathrin mediated endocytosis, and that USP17 depletion impedes plasma membrane recruitment of the machinery required for clathrin mediated endocytosis. Thus, our data reveal that USP17 is necessary for epidermal growth factor receptor and transferrin endocytosis via clathrin coated pits, indicate this is mediated via the regulation of the recruitment of the components of the endocytosis machinery and suggest USP17 may play a general role in receptor endocytosis.

An Event Driven Fusion Approach for Enjoyment Recognition in Real-time

Social signals and interpretation of carried information is of high importance in Human Computer Interaction. Often used for affect recognition, the cues within these signals are displayed in various modalities. Fusion of multi-modal signals is a natural and interesting way to improve automatic classification of emotions transported in social signals. Throughout most present studies, uni-modal affect recognition as well as multi-modal fusion, decisions are forced for fixed annotation segments across all modalities. In this paper, we investigate the less prevalent approach of event driven fusion, which indirectly accumulates asynchronous events in all modalities for final predictions. We present a fusion approach, handling short-timed events in a vector space, which is of special interest for real-time applications. We compare results of segmentation based uni-modal classification and fusion schemes to the event driven fusion approach. The evaluation is carried out via detection of enjoyment-episodes within the audiovisual Belfast Story-Telling Corpus.

Resistance to chemotherapy in ovarian cancer is miRrored by the microRNA miR-433-dependent dysregulation of the cell cycle.

An understanding of the mechanisms underlying the development of resistance to chemotherapy treatment is a gateway to the introduction of novel therapies and improved outcomes for women presenting with ovarian cancer (OC). The desired apoptotic death post-chemotherapy depends on an intact and fully functioning cell cycle machinery.

In this study we demonstrate that stable expression of miR-433 renders OC cells more resistant to paclitaxel treatment. Interestingly, only cells with the highest miR-433 survived paclitaxel suggesting the possible role of miR-433 in cancer recurrence. Importantly, for the first time we demonstrate that miR 433 induces cellular senescence, exemplified by a flattened morphology, the downregulation of phosphorylated Retinoblastoma (p Rb) and increased β galactosidase activity. Surprisingly, miR 433 induced senescence was independent of two well recognised senescent drivers: p21 and p16. Further in silico analysis followed by in vitro experiments identified CKD6 as a novel miR-433 target gene possibly explaining the observed p21 and p16-independent induction of cellular senescence. Another in silico identified miR-433 target gene was CDC27, a protein involved in the regulation of the cell cycle during mitosis. We demonstrate that the overexpression of pre-miR-433 leads to the downregulation of CDC27 in vitro revealing a novel interaction between miR-433 and CDC27, an integral cell cycle regulating protein.

Interestingly, miR-433 expressing cells also demonstrated an ability to impact their tumour microenvironment. We show that miR-433 is present in exosomes released from miR-433 overexpressing and high miR-433 naïve cells. Moreover, growth condition media (GCM) harvested from cells with high miR-433 have higher levels of IL-6 and IL-8, two key cytokines involved in the senescence associated secretory phenotype (SASP). Importantly, GCM from miR-433-enriched cells repressed the growth of co-cultured cells with initial studies showing a GCM-dependent induction of chemoresistance.

In conclusion, data in this study highlights how the aberrant expression miR-433 contributes to chemoresistance in OC cells. We postulate that standard chemotherapy, particularly paclitaxel, used to treat women with OC may have an attenuated ability to kill cells harbouring increased levels of miR-433, allowing for a subsequent chemoresistant phenotype post-therapy.

Hypoxia response element-driven cytosine deaminase/5-fluorocytosine gene therapy system:a highly effective approach to overcome the dynamics of tumour hypoxia and enhance the radiosensitivity of prostate cancer cells in vitro

BACKGROUND: We proposed to exploit hypoxia-inducible factor (HIF)-1alpha overexpression in prostate tumours and use this transcriptional machinery to control the expression of the suicide gene cytosine deaminase (CD) through binding of HIF-1alpha to arrangements of hypoxia response elements. CD is a prodrug activation enzyme, which converts inactive 5-fluorocytosine to active 5-fluorouracil (5-FU), allowing selective killing of vector containing cells.

METHODS: We developed a pair of vectors, containing either five or eight copies of the hypoxia response element (HRE) isolated from the vascular endothelial growth factor (pH5VCD) or glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (pH8GCD) gene, respectively. The kinetics of the hypoxic induction of the vectors and sensitization effects were evaluated in 22Rv1 and DU145 cells in vitro.

RESULTS: The CD protein as selectively detected in lysates of transiently transfected 22Rv1 and DU145 cells following hypoxic exposure. This is the first evidence of GAPDH HREs being used to control a suicide gene therapy strategy. Detectable CD levels were sustained upon reoxygenation and prolonged hypoxic exposures. Hypoxia-induced chemoresistance to 5-FU was overcome in both cell lines treated with this suicide gene therapy approach. Hypoxic transfectants were sensitized to prodrug concentrations that were ten-fold lower than those that are clinically relevant. Moreover, the surviving fraction of reoxygenated transfectants could be further reduced with the concomitant delivery of clinically relevant single radiation doses.

CONCLUSIONS: This strategy thus has the potential to sensitize the hypoxic compartment of prostate tumours and improve the outcome of current therapies.

The emergence of DNA methylation as a key modulator of aberrant cell death in prostate cancer

It is now well established that cancer cells exhibit a number of genetic defects in the machinery that governs programmed cell death and that sabotage of apoptosis is one of the principal factors aiding in the evolution of the carcinogenic phenotype. A number of studies have implicated aberrant DNA methylation as a key survival mechanism in cancer, whereby promoter hypermethylation silences genes essential for many processes including apoptosis. To date, studies on the methylation profile of apoptotic genes have largely focused on cancers of the breast, colon and stomach, with only limited data available on prostate cancer. Here we discuss the major developments in the field of DNA methylation and its role in the regulation of aberrant apoptosis in prostate cancer. The most significant advances have involved the discovery of apoptotic gene targets of methylation, including XAF1, (fragile histidine triad (FHIT ), cellular retinol binding protein 1 (CRBP1), decoy receptor 1(DCR1), decoy receptor 2 (DCR2 ), target of methylation-induced silenceing 1 (TMS1), TNF receptor superfamily, member 6 (FAS), Reprimo (RPRM) and GLI pathogenesis-related 1 (GLIPR1). These genes are reported to be hypermethylated in prostate cancer and some offer potential as diagnostic and prognostic markers. We also introduce the concept of an 'apoptotic methylation signature' for prostate cancer and evaluate its potential in a diagnostic, prognostic and therapeutic setting.

Quantification of Type VI secretion system activity in macrophages infected with Burkholderia cenocepacia

The Gram-negative bacterial type VI Secretion System (T6SS) delivers toxins to kill orinhibit the growth of susceptible bacteria, while others target eukaryotic cells. Deletionof atsR, a negative regulator of virulence factors in B. cenocepacia K56-2, increasesT6SS activity. Macrophages infected with a K56-2 ΔatsR mutant display dramaticalterations in their actin cytoskeleton architecture that rely on the T6SS, which isresponsible for the inactivation of multiple Rho-family GTPases by an unknownmechanism. We employed a strategy to standardize the bacterial infection ofmacrophages and densitometrically quantify the T6SS-associated cellular phenotype,which allowed us to characterize the phenotype of systematic deletions of each genewithin the T6SS cluster and ten vgrG encoding genes in K56-2 ΔatsR. None of thegenes from the T6SS core cluster and the individual vgrGs were directly responsiblefor the cytoskeletal changes in infected cells. However, a mutant strain with all vgrGgenes deleted was unable to cause macrophage alterations. Despite not being able toidentify a specific effector protein responsible for the cytoskeletal defects inmacrophages, our strategy resulted in the identification of the critical core componentsand accessory proteins of the T6SS assembly machinery and provides a screeningmethod to detect T6SS effectors targeting the actin cytoskeleton in macrophages byrandom mutagenesis.

Chromatin binding by the androgen receptor in prostate cancer

Alterations in transcriptional programs are fundamental to the development of cancers. The androgen receptor is central to the normal development of the prostate gland and to the development of prostate cancer. To a large extent this is believed to be due to the control of gene expression through the interaction of the androgen receptor with chromatin and subsequently with coregulators and the transcriptional machinery. Unbiased genome-wide studies have recently uncovered the recruitment sites that are gene-distal and intragenic rather than associated with proximal promoter regions. Whilst expression profiles from AR-positive primary prostate tumours and cell lines can directly relate to the AR cistrome in prostate cancer cells, this distribution raises significant challenges in making direct mechanistic connections. Furthermore, extrapolating from datasets assembled in one model to other model systems or clinical samples poses challenges if we are to use the AR-directed transcriptome to guide the development of novel biomarkers or treatment decisions. This review will provide an overview of the androgen receptor before addressing the challenges and opportunities created by whole-genome studies of the interplay between the androgen receptor and chromatin.

Solitary and repetitive binding motifs for the AP2 complex alpha-appendage in amphiphysin and other accessory proteins

Adaptor protein (AP) complexes bind to transmembrane proteins destined for internalization and to membrane lipids, so linking cargo to the accessory internalization machinery. This machinery interacts with the appendage domains of APs, which have platform and beta-sandwich subdomains, forming the binding surfaces for interacting proteins. Proteins that interact with the subdomains do so via short motifs, usually found in regions of low structural complexity of the interacting proteins. So far, up to four motifs have been identified that bind to and partially compete for at least two sites on each of the appendage domains of the AP2 complex. Motifs in individual accessory proteins, their sequential arrangement into motif domains, and partial competition for binding sites on the appendage domains coordinate the formation of endocytic complexes in a temporal and spatial manner. In this work, we examine the dominant interaction sequence in amphiphysin, a synapse-enriched accessory protein, which generates membrane curvature and recruits the scission protein dynamin to the necks of coated pits, for the platform subdomain of the alpha-appendage. The motif domain of amphiphysin1 contains one copy of each of a DX(F/W) and FXDXF motif. We find that the FXDXF motif is the main determinant for the high affinity interaction with the alpha-adaptin appendage. We describe the optimal sequence of the FXDXF motif using thermodynamic and structural data and show how sequence variation controls the affinities of these motifs for the alpha-appendage.

FCHo proteins are nucleators of clathrin-mediated endocytosis

Clathrin-mediated endocytosis, the major pathway for ligand internalization into eukaryotic cells, is thought to be initiated by the clustering of clathrin and adaptors around receptors destined for internalization. However, here we report that the membrane-sculpting F-BAR domain-containing Fer/Cip4 homology domain-only proteins 1 and 2 (FCHo1/2) were required for plasma membrane clathrin-coated vesicle (CCV) budding and marked sites of CCV formation. Changes in FCHo1/2 expression levels correlated directly with numbers of CCV budding events, ligand endocytosis, and synaptic vesicle marker recycling. FCHo1/2 proteins bound specifically to the plasma membrane and recruited the scaffold proteins eps15 and intersectin, which in turn engaged the adaptor complex AP2. The FCHo F-BAR membrane-bending activity was required, leading to the proposal that FCHo1/2 sculpt the initial bud site and recruit the clathrin machinery for CCV formation.