Identification of eusynstyelamide B as a potent cell cycle inhibitor following the generation and screening of an ascidian-derived extract library using a real time cell analyzer

Ascidians are marine invertebrates that have been a source of numerous cytotoxic compounds. Of the first six marine-derived drugs that made anticancer clinical trials, three originated from ascidian specimens. In order to identify new anti-neoplastic compounds, an ascidian extract library (143 samples) was generated and screened in MDA-MB-231 breast cancer cells using a real-time cell analyzer (RTCA). This resulted in 143 time-dependent cell response profiles (TCRP), which are read-outs of changes to the growth rate, morphology, and adhesive characteristics of the cell culture. Twenty-one extracts affected the TCRP of MDA-MB-231 cells and were further investigated regarding toxicity and specificity, as well as their effects on cell morphology and cell cycle. The results of these studies were used to prioritize extracts for bioassay-guided fractionation, which led to the isolation of the previously identified marine natural product, eusynstyelamide B (1). This bis-indole alkaloid was shown to display an IC50 of 5 μM in MDA-MB-231 cells. Moreover, 1 caused a strong cell cycle arrest in G2/M and induced apoptosis after 72 h treatment, making this molecule an attractive candidate for further mechanism of action studies.

Identification of novel antigens in blood vessels in rectovaginal endometriosis

To identify specific markers of rectovaginal endometriotic nodule vasculature, highly enriched preparations of vascular endothelial cells and pericytes were obtained from endometriotic nodules and control endometrial and myometrial tissue by laser capture microdissection (LCM), and gene expression profiles were screened by microarray analysis. Of the 18 400 transcripts on the arrays, 734 were significantly overexpressed in vessels from fibromuscular tissue and 923 in vessels from stromal tissue of endometriotic nodules, compared with vessels dissected from control tissues. The most frequently expressed transcripts included known endothelial cell-associated genes, as well as transcripts with little or no previous association with vascular cells. The higher expression in blood vessels was further corroborated by immunohistochemical staining of six potential markers, five of which showed strong expression in pericytes. The most promising marker was matrix Gla protein, which was found to be present in both glandular epithelial cells and vascular endothelial cells of endometriotic lesions, although it was barely expressed at all in normal endometrium. LCM, combined with microarray analysis, constitutes a powerful tool for mapping the transcriptome of vascular cells. After immunohistochemical validation, markers of vascular endothelial and perivascular cells from endometriotic nodules could be identified, which may provide targets to improve early diagnosis or to selectively deliver therapeutic agents.

Identification of salivary N-glycoproteins and measurement of glycosylation site occupancy by boronate glycoprotein enrichment and liquid chromatography/electrospray ionization tandem mass spectrometry

RATIONALE Diseases including cancer and congenital disorders of glycosylation have been associated with changes in the site-specific extent of protein glycosylation. Saliva can be non-invasively sampled and is rich in glycoproteins, giving it the potential to be a useful biofluid for the discovery and detection of disease biomarkers associated with changes in glycosylation. METHODS Saliva was collected from healthy individuals and glycoproteins were enriched using phenylboronic acid based glycoprotein enrichment resin. Proteins were deglycosylated with peptide-N-glycosidase F and digested with AspN or trypsin. Desalted peptides and deglycosylated peptides were separated by reversed-phase liquid chromatography and detected with on-line electrospray ionization quadrupole-time-of-flight mass spectrometry using a 5600 TripleTof instrument. Site-specific glycosylation occupancy was semi-quantitatively determined from the abundance of deglycosylated and nonglycosylated versions of each given peptide. RESULTS Glycoprotein enrichment identified 67 independent glycosylation sites from 24 unique proteins, a 3.9-fold increase in the number of glycosylation sites identified. Enrichment of glycoproteins rather than glycopeptides allowed detection of both deglycosylated and nonglycosylated versions of each peptide, and thereby robust measurement of site-specific occupancy at 21 asparagines. Healthy individuals showed limited biological variability in occupancy, with partially modified sites having characteristics consistent with inefficient glycosylation by oligosaccharyltransferase. Inclusion of negative controls without enzymatic deglycosylation controlled for spontaneous chemical deamidation, and identified asparagines previously incorrectly annotated as glycosylated. CONCLUSIONS We developed a sample preparation and mass spectrometry detection strategy for rapid and efficient measurement of site-specific glycosylation occupancy on diverse salivary glycoproteins suitable for biomarker discovery and detection of changes in glycosylation occupancy in human disease.

A little learning is a dangerous thing: Factors influencing the increased identification of special educational needs from the perspective of education policymakers and school practitioners

This article considers the increased identification of special educational needs in Australia’s largest education system from the perspectives of senior public servants, regional directors, principals, school counsellors, classroom teachers, support class teachers, learning support teachers and teaching assistants (n = 30). While their perceptions of an increase generally align with the story told by official statistics, participants’ narratives reveal that school-based identification of special educational needs is neither art nor science. This research finds that rather than an objective indication of the number and nature of children with SEN, official statistics may be more appropriately viewed as a product of funding eligibility and the assumptions of the adults who teach, refer and assess children who experience difficulties in school and with learning.

MIMO and SISO identification of radiation force terms for models of marine structures in waves

This paper presents a discussion on the use of MIMO and SISO techniques for identification of the radiation force terms in models for surface vessels. We compare and discuss two techniques recently proposed in literature for this application: time domain identification and frequency domain identification. We compare the methods in terms of estimates model order, accuracy of the fit, use of the available information, and ease of use and implementation.

Identification of pre-spike network in patients with mesial temporal lobe epilepsy

Background: Seizures and interictal spikes in mesial temporal lobe epilepsy (MTLE) affect a network of brain regions rather than a single epileptic focus. Simultaneous electroencephalography and functional magnetic resonance imaging (EEG-fMRI) studies have demonstrated a functional network in which hemodynamic changes are time-locked to spikes. However, whether this reflects the propagation of neuronal activity from a focus, or conversely the activation of a network linked to spike generation remains unknown. The functional connectivity (FC) changes prior to spikes may provide information about the connectivity changes that lead to the generation of spikes. We used EEG-fMRI to investigate FC changes immediately prior to the appearance of interictal spikes on EEG in patients with MTLE. Methods/principal findings: Fifteen patients with MTLE underwent continuous EEG-fMRI during rest. Spikes were identified on EEG and three 10 s epochs were defined relative to spike onset: spike (0–10 s), pre-spike (−10 to 0 s), and rest (−20 to −10 s, with no previous spikes in the preceding 45s). Significant spike-related activation in the hippocampus ipsilateral to the seizure focus was found compared to the pre-spike and rest epochs. The peak voxel within the hippocampus ipsilateral to the seizure focus was used as a seed region for FC analysis in the three conditions. A significant change in FC patterns was observed before the appearance of electrographic spikes. Specifically, there was significant loss of coherence between both hippocampi during the pre-spike period compared to spike and rest states. Conclusion/significance: In keeping with previous findings of abnormal inter-hemispheric hippocampal connectivity in MTLE, our findings specifically link reduced connectivity to the period immediately before spikes. This brief decoupling is consistent with a deficit in mutual (inter-hemispheric) hippocampal inhibition that may predispose to spike generation.

Nanostructured silver-gold bimetallic SERS substrates for selective identification of bacteria in human blood

Surface-enhanced Raman spectroscopy (SERS) is a potentially important tool in the rapid and accurate detection of pathogenic bacteria in biological fluids. However, for diagnostic application of this technique, it is necessary to develop a highly sensitive, stable, biocompatible and reproducible SERS-active substrate. In this work, we have developed a silver–gold bimetallic SERS surface by a simple potentiostatic electrodeposition of a thin gold layer on an electrochemically roughened nanoscopic silver substrate. The resultant substrate was very stable under atmospheric conditions and exhibited the strong Raman enhancement with the high reproducibility of the recorded SERS spectra of bacteria (E. coli, S. enterica, S. epidermidis, and B. megaterium). The coating of the antibiotic over the SERS substrate selectively captured bacteria from blood samples and also increased the Raman signal in contrast to the bare surface. Finally, we have utilized the antibiotic-coated hybrid surface to selectively identify different pathogenic bacteria, namely E. coli, S. enterica and S. epidermidis from blood samples.

Wrecking ball: could big tech firms ruin the Spotify party?

YouTube is contemplating the launch of a new music service. But how would such a service fare against established music services like Spotify, Rdio, and Pandora? All these services are referred to as “access-based music services”. They offer music listeners access to millions of songs they can listen to as much as they want for free (with advertising and only basic functionality) or for a monthly subscription fee (without advertising)...

Superstars as drivers of organizational identification : empirical findings from professional soccer

This paper examines the effect of superstars on external stakeholders’ organizational identification through the lens of sport. Drawing on social identity theory and the concept of organizational identification, as well as on role model theories and superstar economics, several hypotheses are developed regarding the influence of soccer stars on their fans’ degree of team identification. Using a proprietary data set that combines archival data on professional German soccer players and clubs with survey data on more than 1,400 soccer fans, this study finds evidence for a positive effect of superstar characteristics and role model perception. Moreover, it is found that players who qualify for the definition of a superstar are more important to fans of established teams than to fans of unsuccessful teams. The player's club tenure, however, seems to have no influence on fans’ team identification. It is further argued that the effect of soccer stars on their fans is comparable to that of executives on external stakeholders, and hence, the results are applied to the business domain. The results of this study contribute to existing research by extending the list of personnel-related determinants of organizational identification.

Text mining with semantic annotation : using enriched text representation for entity-oriented retrieval, semantic relation identification and text clustering

This project is a step forward in the study of text mining where enhanced text representation with semantic information plays a significant role. It develops effective methods of entity-oriented retrieval, semantic relation identification and text clustering utilizing semantically annotated data. These methods are based on enriched text representation generated by introducing semantic information extracted from Wikipedia into the input text data. The proposed methods are evaluated against several start-of-art benchmarking methods on real-life data-sets. In particular, this thesis improves the performance of entity-oriented retrieval, identifies different lexical forms for an entity relation and handles clustering documents with multiple feature spaces.

Causal factors of hot air ballooning incidents : identification, frequency, and potential impact

Background: Hot air ballooning incidents are relatively rare, however, when they do occur they are likely to result in a fatality or serious injury. Human error is commonly attributed as the cause of hot air ballooning incidents; however, error in itself is not an explanation for safety failures. This research aims to identify, and establish the relative importance of factors contributing towards hot air ballooning incidents. Methods: Twenty-two Australian Ballooning Federation (ABF) incident reports were thematically coded using a bottom up approach to identify causal factors. Subsequently, 69 balloonists (mean 19.51 years’ experience) participated in a survey to identify additional causal factors and rate (out of seven) the perceived frequency and potential impact to ballooning operations of each of the previously identified causal factors. Perceived associated risk was calculated by multiplying mean perceived frequency and impact ratings. Results: Incident report coding identified 54 causal factors within nine higher level areas: Attributes, Crew resource management, Equipment, Errors, Instructors, Organisational, Physical Environment, Regulatory body and Violations. Overall, ‘weather’, ‘inexperience’ and ‘poor/inappropriate decisions’ were rated as having greatest perceived associated risk. Discussion: Although errors were nominated as a prominent cause of hot air ballooning incidents, physical environment and personal attributes are also particularly important for safe hot air ballooning operations. In identifying a range of causal factors the areas of weakness surrounding ballooning operations have been defined; it is hoped that targeted safety and training strategies can now be put into place removing these contributing factors and reducing the chance of pilot error.

Correlation based method for phase identification in a three phase LV distribution network

Low voltage distribution networks feature a high degree of load unbalance and the addition of rooftop photovoltaic is driving further unbalances in the network. Single phase consumers are distributed across the phases but even if the consumer distribution was well balanced when the network was constructed changes will occur over time. Distribution transformer losses are increased by unbalanced loadings. The estimation of transformer losses is a necessary part of the routine upgrading and replacement of transformers and the identification of the phase connections of households allows a precise estimation of the phase loadings and total transformer loss. This paper presents a new technique and preliminary test results for a method of automatically identifying the phase of each customer by correlating voltage information from the utility's transformer system with voltage information from customer smart meters. The techniques are novel as they are purely based upon a time series of electrical voltage measurements taken at the household and at the distribution transformer. Experimental results using a combination of electrical power and current of the real smart meter datasets demonstrate the performance of our techniques.

Consumer phase identification in a three phase unbalanced LV distribution network

A new technique is presented for automatically identifying the phase connection of domestic customers. Voltage information from a reference three phase house is correlated with voltage information from other customer electricity meters on the same network to determine the highest probability phase connection. The techniques are purely based upon a time series of electrical voltage measurements taken by the household smart meters and no additional equipment is required. The method is demonstrated using real smart meter datasets to correctly identify the phase connections of 75 consumers on a low voltage distribution feeder.

Identification of allanite (Ce, Ca, Y)2(Al, Fe3+)3(SiO4)3OH found in marble from Chillagoe, Queensland using Raman spectroscopy

Samples of marble from Chillagoe, North Queensland have been analyzed using scanning electron microscopy (SEM) with energy dispersive X-ray spectroscopy (EDS) and Raman spectroscopy. Chemical analyses provide evidence for the presence of minerals other than limestone and calcite in the marble, including silicate minerals. Some of these analyses correspond to silicate minerals. The Raman spectra of these crystals were obtained and the Raman spectrum corresponds to that of allanite from the Arizona State University data base (RRUFF) data base. The combination of SEM with EDS and Raman spectroscopy enables the characterization of the mineral allanite in the Chillagoe marble.

A continental-scale tool for acoustic identification of European bats

Summary 1. Acoustic methods are used increasingly to survey and monitor bat populations. However, the use of acoustic methods at continental scales can be hampered by the lack of standardized and objective methods to identify all species recorded. This makes comparable continent-wide monitoring difficult, impeding progress towards developing biodiversity indicators, transboundary conservation programmes and monitoring species distribution changes. 2. Here we developed a continental-scale classifier for acoustic identification of bats, which can be used throughout Europe to ensure objective, consistent and comparable species identifications. We selected 1350 full-spectrum reference calls from a set of 15 858 calls of 34 European species, from EchoBank, a global echolocation call library. We assessed 24 call parameters to evaluate how well they distinguish between species and used the 12 most useful to train a hierarchy of ensembles of artificial neural networks to distinguish the echolocation calls of these bat species. 3. Calls are first classified to one of five call-type groups, with a median accuracy of 97·6%. The median species-level classification accuracy is 83·7%, providing robust classification for most European species, and an estimate of classification error for each species. 4. These classifiers were packaged into an online tool, iBatsID, which is freely available, enabling anyone to classify European calls in an objective and consistent way, allowing standardized acoustic identification across the continent. 5. Synthesis and applications. iBatsID is the first freely available and easily accessible continental- scale bat call classifier, providing the basis for standardized, continental acoustic bat monitoring in Europe. This method can provide key information to managers and conservation planners on distribution changes and changes in bat species activity through time.