Single live-cell imaging for systems biology

Understanding how mammalian cells function requires a dynamic perspective. However, due to the complexity of signalling networks these non-linear systems can easily elude human intuition. The central aim of systems biology is to improve our understanding of the temporal complexity of cell signalling pathways, using a combination of experimental and computational approaches. Live cell imaging and computational modelling are compatible techniques which allow quantitative analysis of cell signalling pathway dynamics. Non-invasive imaging techniques, based on the use of various luciferases and fluorescent proteins, trace cellular events such as gene expression, protein-protein interactions and protein localisation in cells. By employing a number of markers in a single assay, multiple parameters can be measured simultaneously in the same cell. Following acquisition using specialised microscopy, analysis of multi-parameter time-lapse images facilitates the identification of important qualitative and quantitative relationships – linking intracellular signalling, gene expression and cell fate. Improvements in reporter genes coupled with significant advances in detector technologies, are now allowing us to image gene expression non-invasively in individual living cells. These methods are providing remarkable insights into the dynamics of gene expression during complex processes, such as the cell cycle and the responses of cells to hormones, growth factors and nutrients. On a larger scale, dynamics of gene expression may also be monitored in living organisms. This new technology will greatly assist attempts to decipher the complex behaviours exhibited by biological signalling networks, for instance the ability to integrate multiple input signals over time, and generate specific outputs

InterPACK2011-52268 USE CONDITION CHARACTERIZATION OF PACKAGE COMPONENTS

ABSTRACT Discrete components, such as capacitors and inductors, play an important role in the analysis and design of electronic packages and printed circuit boards. Although the electrical parameters of discrete components are described by manufacturers, the component performance at product operating conditions can vary drastically from the manufacturer's specification. Accurate characterization of discrete package components at operating conditions is essential to understand product operation. This paper will introduce a method to characterize discrete capacitors and inductors while applying multiple operating conditions simultaneously. Several inductor options will be evaluated, including a newly introduced metal composite component

Effect of Exercise on Health-Related Quality of Life in Patients with End-Stage Renal Disease

Chronic kidney disease (CKD) is becoming more common around the world. Chronic kidney disease (CKD) is linked to a wide range of other health problems, such as diabetes, hypertension, stroke, and pulmonary illness. Patients with CKD tend to lead sedentary lives for a variety of reasons. Dialysis patients, on the other hand, are much less active than the general population. All of these factors raise the likelihood of future morbidity and mortality, while also lowering the overall quality of life for people who are ill (HRQoL). Regular physical activity (PE) has been shown to increase overall well-being and HRQoL. Here, we discuss several PEs and their effects on CKD patients\u27 physical fitness, function, and HRQoL, as well as the significance of haematocrit normalisation and the influence on their serum phosphorus levels. We have discussed the advantages of PE for this particular population of individuals as well as the side effects of intradialytic PE. There have also been discussions on factors that contribute to impaired physical function in CKD patients and the impact of PEs on different bodily systems

Stage 4S Neuroblastoma: What Are the Outcomes? A Systematic Review of Published Studies

Introduction The prognosis of stage 4S/MS neuroblastoma has traditionally been reported as excellent, yet conflicting treatment protocols exist for this enigmatic disease. To critically address this question, we have undertaken a systematic review of published studies to accurately determine outcomes for infants with stage 4S/MS neuroblastoma.Materials and Methods Studies were identified using MEDLINE, Embase, and Cochrane databases using the relevant search terms. Literature reviews, case reports, and adult studies were excluded. Data were extracted independently following article selection by three authors and reviewed by the senior author.Results The original search retrieved 2,325 articles. Following application of exclusion criteria and removing duplicate data, 37 studies (1,105 patients) were included for final review. Overall patient survival was 84%. Twelve studies (544 patients) recorded MYCN status. Mortality in MYCN amplified tumors was 56%. Chromosome 1p/11q status was reported in four studies and 1p/11q deletion carried a 40% fatality rate. Management included observation only (201 patients, 8.5% mortality), surgical resection of primary tumor only (153 patients, 6.5% mortality), chemotherapy only (186 patients, 21% mortality), radiotherapy (5 deaths, 33% mortality), chemotherapy with surgery (160 patients, 10% mortality), surgery with radiotherapy (21 patients, 19% mortality), radiotherapy with chemotherapy (42 patients, 29% mortality), and surgery with chemotherapy and radiotherapy (27 patients, 33% mortality).Conclusion There is a significant mortality observed in stage 4S/MS neuroblastoma infants with a dismal outcome observed in those patients with MYCN amplification and 1p/11q deletion. Those patients suitably amenable for conservative management or surgery to excise the primary tumor carry the best prognosis.</p

Global millimeter VLBI array survey of ultracompact extragalactic radio sources at 86 GHz

Context. Very long baseline interferometry (VLBI) observations at 86 GHz (wavelength, λ = 3 mm) reach a resolution of about 50 μas, probing the collimation and acceleration regions of relativistic outflows in active galactic nuclei (AGN). The physical conditions in these regions can be studied by performing 86 GHz VLBI surveys of representative samples of compact extragalactic radio sources. Aims: To extend the statistical studies of compact extragalactic jets, a large global 86 GHz VLBI survey of 162 compact radio sources was conducted in 2010-2011 using the Global Millimeter VLBI Array (GMVA). Methods: The survey observations were made in a snapshot mode, with up to five scans per target spread over a range of hour angles in order to optimize the visibility coverage. The survey data attained a typical baseline sensitivity of 0.1 Jy and a typical image sensitivity of 5 mJy beam-1, providing successful detections and images for all of the survey targets. For 138 objects, the survey provides the first ever VLBI images made at 86 GHz. Gaussian model fitting of the visibility data was applied to represent the structure of the observed sources and to estimate the flux densities and sizes of distinct emitting regions (components) in their jets. These estimates were used for calculating the brightness temperature (Tb) at the jet base (core) and in one or more moving regions (jet components) downstream from the core. These model-fit-based estimates of Tb were compared to the estimates of brightness temperature limits made directly from the visibility data, demonstrating a good agreement between the two methods. Results: The apparent brightness temperature estimates for the jet cores in our sample range from 2.5 × 109 K to 1.3 × 1012 K, with the mean value of 1.8 × 1011 K. The apparent brightness temperature estimates for the inner jet components in our sample range from 7.0 × 107 K to 4.0 × 1011 K. A simple population model with a single intrinsic value of brightness temperature, T0, is applied to reproduce the observed distribution. It yields T0 = (3.77-0.14+0.10) × 1011 K for the jet cores, implying that the inverse Compton losses dominate the emission. In the nearest jet components, T0 = (1.42-0.19+0.16) × 1011 K is found, which is slightly higher than the equipartition limit of ̃5 × 1010 K expected for these jet regions. For objects with sufficient structural detail detected, the adiabatic energy losses are shown to dominate the observed changes of brightness temperature along the jet. The reduced images and visibility tables (FITS files) and the full Tables 5-7 are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz- bin/qcat?J/A+A/622/A9

Plastin and spectrin cooperate to stabilize the actomyosin cortex during cytokinesis

Cytokinesis, the process that partitions the mother cell into two daughter cells, requires the assembly and constriction of an equatorial actomyosin network. Different types of non-motor F-actin crosslinkers localize to the network, but their functional contribution remains poorly understood. Here, we describe a synergy between the small rigid crosslinker plastin and the large flexible crosslinker spectrin in the C. elegans one-cell embryo. In contrast to single inhibitions, co-inhibition of plastin and the βH-spectrin (SMA-1) results in cytokinesis failure due to progressive disorganization and eventual collapse of the equatorial actomyosin network. Cortical localization dynamics of non-muscle myosin II in co-inhibited embryos mimic those observed after drug-induced F-actin depolymerization, suggesting that the combined action of plastin and spectrin stabilizes F-actin in the contractile ring. An in silico model predicts that spectrin is more efficient than plastin at stabilizing the ring and that ring formation is relatively insensitive to βH-spectrin length, which is confirmed in vivo with a sma-1 mutant that lacks 11 of its 29 spectrin repeats. Our findings provide the first evidence that spectrin contributes to cytokinesis and highlight the importance of crosslinker interplay for actomyosin network integrity

Mucin (Muc) expression during pancreatic cancer progression in spontaneous mouse model: potential implications for diagnosis and therapy.

BACKGROUND: Pancreatic cancer (PC) is a lethal malignancy primarily driven by activated Kras mutations and characterized by the deregulation of several genes including mucins. Previous studies on mucins have identified their significant role in both benign and malignant human diseases including PC progression and metastasis. However, the initiation of MUC expression during PC remains unknown because of lack of early stage tumor tissues from PC patients. METHODS: In the present study, we have evaluated stage specific expression patterns of mucins during mouse PC progression in (Kras(G12D);Pdx1-Cre (KC)) murine PC model from pancreatic intraepithelial neoplasia (PanIN) to pancreatic ductal adenocarcinoma (PDAC) by immunohistochemistry and quantitative real-time PCR. RESULTS: In agreement with previous studies on human PC, we observed a progressive increase in the expression of mucins particularly Muc1, Muc4 and Muc5AC in the pancreas of KC (as early as PanIN I) mice with advancement of PanIN lesions and PDAC both at mRNA and protein levels. Additionally, mucin expression correlated with the increased expression of inflammatory cytokines IFN-γ (p \u3c 0.0062), CXCL1 (p \u3c 0.00014) and CXCL2 (p \u3c 0.08) in the pancreas of KC mice, which are known to induce mucin expression. Further, we also observed progressive increase in inflammation in pancreas of KC mice from 10 to 50 weeks of age as indicated by the increase in the macrophage infiltration. Overall, this study corroborates with previous human studies that indicated the aberrant overexpression of MUC1, MUC4 and MUC5AC mucins during the progression of PC. CONCLUSIONS: Our study reinforces the potential utility of the KC murine model for determining the functional role of mucins in PC pathogenesis by crossing KC mice with corresponding mucin knockout mice and evaluating mucin based diagnostic and therapeutic approaches for lethal PC

Amyloid precursor-like protein 2 (APLP2) affects the actin cytoskeleton and increases pancreatic cancer growth and metastasis.

Amyloid precursor-like protein 2 (APLP2) is aberrantly expressed in pancreatic cancer. Here we showed that APLP2 is increased in pancreatic cancer metastases, particularly in metastatic lesions found in the diaphragm and intestine. Examination of matched human primary tumor-liver metastasis pairs showed that 38.1% of the patients had positive APLP2 expression in both the primary tumor and the corresponding liver metastasis. Stable knock-down of APLP2 expression (with inducible shRNA) in pancreatic cancer cells reduced the ability of these cells to migrate and invade. Loss of APLP2 decreased cortical actin and increased intracellular actin filaments in pancreatic cancer cells. Down-regulation of APLP2 decreased the weight and metastasis of orthotopically transplanted pancreatic tumors in nude mice

Pathobiological Implications of MUC16 Expression in Pancreatic Cancer

MUC16 (CA125) belongs to a family of high-molecular weight O-glycosylated proteins known as mucins. While MUC16 is well known as a biomarker in ovarian cancer, its expression pattern in pancreatic cancer (PC), the fourth leading cause of cancer related deaths in the United States, remains unknown. The aim of our study was to analyze the expression of MUC16 during the initiation, progression and metastasis of PC for possible implication in PC diagnosis, prognosis and therapy. In this study, a microarray containing tissues from healthy and PC patients was used to investigate the differential protein expression of MUC16 in PC. MUC16 mRNA levels were also measured by RT-PCR in the normal human pancreatic, pancreatitis, and PC tissues. To investigate its expression pattern during PC metastasis, tissue samples from the primary pancreatic tumor and metastases (from the same patient) in the lymph nodes, liver, lung and omentum from Stage IV PC patients were analyzed. To determine its association in the initiation of PC, tissues from PC patients containing pre-neoplastic lesions of varying grades were stained for MUC16. Finally, MUC16 expression was analyzed in 18 human PC cell lines. MUC16 is not expressed in the normal pancreatic ducts and is strongly upregulated in PC and detected in pancreatitis tissue. It is first detected in the high-grade pre-neoplastic lesions preceding invasive adenocarcinoma, suggesting that its upregulation is a late event during the initiation of this disease. MUC16 expression appears to be stronger in metastatic lesions when compared to the primary tumor, suggesting a role in PC metastasis. We have also identified PC cell lines that express MUC16, which can be used in future studies to elucidate its functional role in PC. Altogether, our results reveal that MUC16 expression is significantly increased in PC and could play a potential role in the progression of this disease