Aβ(1-42) induced metabolic effects in a stem cell derived neuron and astrocyte network.

Alzheimer’s Disease (AD) is the most common form of dementia currently affecting more than 35 million people worldwide. Hypometabolism is a major feature of AD and appears decades before cognitive decline and pathological lesions. This has a detrimental impact on the brain which has a high energy demand. Current models of AD fail to mimic all the features of the disease, which has an impact on the development of new therapies. Human stem cell derived models of the brain have attracted a lot of attention in recent years as a tool to study neurodegenerative diseases. In this thesis, neurons and astrocytes derived from the human embryonal carcinoma cell line (NT2/D1) were utilised to determine the metabolic coupling between neurons and astrocytes with regards to responses to hypoglycaemia, neuromodulators and increase in neuronal activity. This model was then used to investigate the effects of Aß(1-42) on the metabolism of these NT2-derived co-cultures as well as pure astrocytes. Additionally primary cortical mixed neuronal and glial cultures were utilised to compare this model to a widely accepted in vitro model used in Alzheimer’s disease research. Co-cultures were found to respond to Aß(1-42) in similar way to human and in vivo models. Hypometabolism was characterised by changes in glucose metabolism, as well as lactate, pyruvate and glycogen. This led to a significant decrease in ATP and the ratio of NAD+/NADH. These results together with an increase in calcium oscillations and a decrease in GSH/GSSG ratio, suggests Aß-induces metabolic and oxidative stress. This situation could have detrimental effects in the brain which has a high energy demand, especially in terms of memory formation and antioxidant capacity.

Gene network and proteomic analyses of cardiac responses to pathological and physiological stress

Background—The molecular mechanisms underlying similarities and differences between physiological and pathological left ventricular hypertrophy (LVH) are of intense interest. Most previous work involved targeted analysis of individual signaling pathways or screening of transcriptomic profiles. We developed a network biology approach using genomic and proteomic data to study the molecular patterns that distinguish pathological and physiological LVH. Methods and Results—A network-based analysis using graph theory methods was undertaken on 127 genome-wide expression arrays of in vivo murine LVH. This revealed phenotype-specific pathological and physiological gene coexpression networks. Despite >1650 common genes in the 2 networks, network structure is significantly different. This is largely because of rewiring of genes that are differentially coexpressed in the 2 networks; this novel concept of differential wiring was further validated experimentally. Functional analysis of the rewired network revealed several distinct cellular pathways and gene sets. Deeper exploration was undertaken by targeted proteomic analysis of mitochondrial, myofilament, and extracellular subproteomes in pathological LVH. A notable finding was that mRNA–protein correlation was greater at the cellular pathway level than for individual loci. Conclusions—This first combined gene network and proteomic analysis of LVH reveals novel insights into the integrated pathomechanisms that distinguish pathological versus physiological phenotypes. In particular, we identify differential gene wiring as a major distinguishing feature of these phenotypes. This approach provides a platform for the investigation of potentially novel pathways in LVH and offers a freely accessible protocol (http://sites.google.com/site/cardionetworks) for similar analyses in other cardiovascular diseases.

Intra-channel nonlinearity compensation for PM-16QAM traffic co-propagating with 28Gbaud m-ary QAM neighbours

We quantify the benefits of intra-channel nonlinear compensation in meshed optical networks, in view of network configuration, fibre design aspect, and dispersion management. We report that for a WDM optical transport network employing flexible 28Gbaud PM-mQAM transponders with no in-line dispersion compensation, intrachannel nonlinear compensation, for PM-16QAM through traffic, offers significant improvements of up to 4dB in nonlinear tolerance (Q-factor) irrespective of the co-propagating modulation format, and that this benefit is further enhanced (1.5dB) by increasing local link dispersion. For dispersion managed links, we further report that advantages of intra-channel nonlinear compensation increase with in-line dispersion compensation ratio, with 1.5dB improvements after 95% in-line dispersion compensation, compared to uncompensated transmission. © 2012 Optical Society of America.

Managing supply networks:organizational roles in network management

This paper describes research findings on the roles that organizations can adopt in managing supply networks. Drawing on extensive empirical data, it is demonstrated that organizations may be said to be able to manage supply networks, provided a broad view of ‘managing’ is adopted. Applying role theory, supply network management interventions were clustered into sets of linked activities and goals that constituted supply network management roles. Six supply network management roles were identified – innovation facilitator, co-ordinator, supply policy maker and implementer, advisor, information broker and supply network structuring agent. The findings are positioned in the wider context of debates about the meaning of management, the contribution of role theory to our understanding of management, and whether inter-organizational networks can be managed.

Supply network strategy:role and competence requirements

This paper presents an argument that it is possible for an organisation to manage networks, but understanding this involves consideration of what is meant by "managing". Based on prior research and data from a major longitudinal action research study in the health sector, the paper describes six network management roles: network structuring agent; co-ordinator; advisor; information broker; relationship broker; innovation sponsor. The necessary "assets" for effective performance of these roles are identified, in particular those relating to team competence. The findings enrich and significantly develop previous work on network management roles and activities, and their influencing factors. It is concluded that, given the specific nature of the networks studied, further research is required to evaluate the generalisability of the findings, though initial indications are promising.

Quantification of functionalised gold nanoparticle-targeted knockdown of gene expression in HeLa cells

Introduction: Gene therapy continues to grow as an important area of research, primarily because of its potential in the treatment of disease. One significant area where there is a need for better understanding is in improving the efficiency of oligonucleotide delivery to the cell and indeed, following delivery, the characterization of the effects on the cell. Methods: In this report, we compare different transfection reagents as delivery vehicles for gold nanoparticles functionalized with DNA oligonucleotides, and quantify their relative transfection efficiencies. The inhibitory properties of small interfering RNA (siRNA), single-stranded RNA (ssRNA) and single-stranded DNA (ssDNA) sequences targeted to human metallothionein hMT-IIa are also quantified in HeLa cells. Techniques used in this study include fluorescence and confocal microscopy, qPCR and Western analysis. Findings: We show that the use of transfection reagents does significantly increase nanoparticle transfection efficiencies. Furthermore, siRNA, ssRNA and ssDNA sequences all have comparable inhibitory properties to ssDNA sequences immobilized onto gold nanoparticles. We also show that functionalized gold nanoparticles can co-localize with autophagosomes and illustrate other factors that can affect data collection and interpretation when performing studies with functionalized nanoparticles. Conclusions: The desired outcome for biological knockdown studies is the efficient reduction of a specific target; which we demonstrate by using ssDNA inhibitory sequences targeted to human metallothionein IIa gene transcripts that result in the knockdown of both the mRNA transcript and the target protein. © 2014 Jiwaji et al.

Neuronal network dynamics during epileptogenesis in the medial temporal lobe

Epilepsy is one of the most common neurological disorders, a large fraction of which is resistant to pharmacotherapy. In this light, understanding the mechanisms of epilepsy and its intractable forms in particular could create new targets for pharmacotherapeutic intervention. The current project explores the dynamic changes in neuronal network function in the chronic temporal lobe epilepsy (TLE) in rat and human brain in vitro. I focused on the process of establishment of epilepsy (epileptogenesis) in the temporal lobe. Rhythmic behaviour of the hippocampal neuronal networks in healthy animals was explored using spontaneous oscillations in the gamma frequency band (SγO). The use of an improved brain slice preparation technique resulted in the natural occurence (in the absence of pharmacological stimulation) of rhythmic activity, which was then pharmacologically characterised and compared to other models of gamma oscillations (KA- and CCh-induced oscillations) using local field potential recording technique. The results showed that SγO differed from pharmacologically driven models, suggesting higher physiological relevance of SγO. Network activity was also explored in the medial entorhinal cortex (mEC), where spontaneous slow wave oscillations (SWO) were detected. To investigate the course of chronic TLE establishment, a refined Li-pilocarpine-based model of epilepsy (RISE) was developed. The model significantly reduced animal mortality and demonstrated reduced intensity, yet high morbidy with almost 70% mean success rate of developing spontaneous recurrent seizures. We used SγO to characterize changes in the hippocampal neuronal networks throughout the epileptogenesis. The results showed that the network remained largely intact, demonstrating the subtle nature of the RISE model. Despite this, a reduction in network activity was detected during the so-called latent (no seizure) period, which was hypothesized to occur due to network fragmentation and an abnormal function of kainate receptors (KAr). We therefore explored the function of KAr by challenging SγO with kainic acid (KA). The results demonstrated a remarkable decrease in KAr response during the latent period, suggesting KAr dysfunction or altered expression, which will be further investigated using a variety of electrophysiological and immunocytochemical methods. The entorhinal cortex, together with the hippocampus, is known to play an important role in the TLE. Considering this, we investigated neuronal network function of the mEC during epileptogenesis using SWO. The results demonstrated a striking difference in AMPAr function, with possible receptor upregulation or abnormal composition in the early development of epilepsy. Alterations in receptor function inevitably lead to changes in the network function, which may play an important role in the development of epilepsy. Preliminary investigations were made using slices of human brain tissue taken following surgery for intratctable epilepsy. Initial results showed that oscillogenesis could be induced in human brain slices and that such network activity was pharmacologically similar to that observed in rodent brain. Overall, our findings suggest that excitatory glutamatergic transmission is heavily involved in the process of epileptogenesis. Together with other types of receptors, KAr and AMPAr contribute to epilepsy establishment and may be the key to uncovering its mechanism.

Expression of VEGF-C and activation of its receptors VEGFR-2 and VEGFR-3 in trophoblast

Placental villous development requires the co-ordinated action of angiogenic factors on both endothelial and trophoblast cells. Like vascular endothelial growth factor (VEGF), VEGF-C increases vascular permeability, stimulates endothelial cell proliferation and migration. In the present study, we investigated the expression of VEGF-C and its receptors VEGFR-3 and VEGFR-2 in normal and intrauterine growth-restricted (IUGR) placenta. Immunolocalisation studies showed that like VEGF and VEGFR-1, VEGF-C, VEGFR-3 and VEGFR-2 co-localised to the syncytiotrophoblast, to cells in the maternal decidua, as well as to the endothelium of the large placental blood vessels. Western blot analysis demonstrated a significant decrease in placental VEGF-C and VEGFR-3 protein expression in severe IUGR as compared to gestationally-matched third trimester pregnancies. Conditioned medium from VEGF-C producing pancreatic carcinoma (Suit-2) and endometrial epithelial (Hec-1B) cell lines caused an increased association of the phosphorylated extracellular signal regulated kinase (ERK) in VEGFR-3 immunoprecipitates from spontaneously transformed first trimester trophoblast cells. VEGF121 caused dose-dependant phosphorylation of VEGFR-2 in trophoblast cells as well as stimulating DNA synthesis. In addition, premixing VEGF165 with heparin sulphate proteoglycan potentiated trophoblast proliferation and the association of phospho-ERK with the VEGFR-2 receptor. VEGF165-mediated DNA synthesis was inhibited by anti-VEGFR-2 neutralising antibody. The results demonstrate functional VEGFR-2 and VEGFR-3 receptors on trophoblast and suggest that the decreased expression of VEGF-C and VEGFR-3 may contribute to the abnormal villous development observed in IUGR placenta.

A comparison of two distributed large-volume measurement systems:the mobile spatial co-ordinate measuring system and the indoor global positioning system

Advances in the area of industrial metrology have generated new technologies that are capable of measuring components with complex geometry and large dimensions. However, no standard or best-practice guides are available for the majority of such systems. Therefore, these new systems require appropriate testing and verification in order for the users to understand their full potential prior to their deployment in a real manufacturing environment. This is a crucial stage, especially when more than one system can be used for a specific measurement task. In this paper, two relatively new large-volume measurement systems, the mobile spatial co-ordinate measuring system (MScMS) and the indoor global positioning system (iGPS), are reviewed. These two systems utilize different technologies: the MScMS is based on ultrasound and radiofrequency signal transmission and the iGPS uses laser technology. Both systems have components with small dimensions that are distributed around the measuring area to form a network of sensors allowing rapid dimensional measurements to be performed in relation to large-size objects, with typical dimensions of several decametres. The portability, reconfigurability, and ease of installation make these systems attractive for many industries that manufacture large-scale products. In this paper, the major technical aspects of the two systems are briefly described and compared. Initial results of the tests performed to establish the repeatability and reproducibility of these systems are also presented. © IMechE 2009.

Functional astrocyte-neuron lactate shuttle in a human stem cell derived neuronal network

The development of stem cell-derived neuronal networks will promote experimental system development for drug screening, toxicological testing and disease modelling, providing that they mirror closely the functional competencies of their in vivo counterparts. The NT2 cell line is one of the best documented embryocarcinoma cell lines, and can be differentiated into neurons and astrocytes. Great focus has also been placed on defining the electrophysiological properties of these cells, and more recently we have investigated the functional properties of their associated astrocytes. We now show for the first time in a human stem cell derived co-culture model that these cultures are also metabolically competent and demonstrate a functional astrocyte neuron lactate shuttle (ANLS). The ANLS hypothesis proposes that during neuronal activity, glutamate released into the synaptic cleft is taken up by astrocytes and triggers glucose uptake which is converted into lactate and released via monocarboxylate transporters for neuronal use. Using mixed cultures of NT2 derived neurons and astrocytes we have shown that these cells modulate their glucose uptake in response to glutamate, an effect that was blocked by cytochalasin B and ouabain. Additionally we demonstrate that in response to increased neuronal activity and under hypoglycaemic conditions, co-cultures modulate glycogen turnover and increase lactate production. Similar results were also shown following treatment with glutamate, potassium, Isoproterenol and dbcAMP. Together these results demonstrate for the first time a functional ANLS in a human stem cell derived co-culture.

Numerical analysis of artificial neural network and volterra-based nonlinear equalizers for coherent optical OFDM

One major drawback of coherent optical orthogonal frequency-division multiplexing (CO-OFDM) that hitherto remains unsolved is its vulnerability to nonlinear fiber effects due to its high peak-to-average power ratio. Several digital signal processing techniques have been investigated for the compensation of fiber nonlinearities, e.g., digital back-propagation, nonlinear pre- and post-compensation and nonlinear equalizers (NLEs) based on the inverse Volterra-series transfer function (IVSTF). Alternatively, nonlinearities can be mitigated using nonlinear decision classifiers such as artificial neural networks (ANNs) based on a multilayer perceptron. In this paper, ANN-NLE is presented for a 16QAM CO-OFDM system. The capability of the proposed approach to compensate the fiber nonlinearities is numerically demonstrated for up to 100-Gb/s and over 1000km and compared to the benchmark IVSTF-NLE. Results show that in terms of Q-factor, for 100-Gb/s at 1000km of transmission, ANN-NLE outperforms linear equalization and IVSTF-NLE by 3.2dB and 1dB, respectively.

Networked governance and the regulation of expression on the Internet:the blurring of the role of public and private actors as content regulators

This editorial provides an overview of the themes of network governance and content regulation that are expanded upon in the subsequent articles, identifying key issues and concerns that are prevalent in the literature in this field. In particular, this text considers governance not as an Internet-specific phenomenon, but as a global phenomenon, identifying and discussing literature pertaining to governance both online and offline, and providing examples of theories that seek to explain these forms of governance. Focusing on the interaction between public and private actors in content regulation, this editorial highlights that content regulation is a complex and contested issue that cannot be separated from its social and cultural contexts, and provides an overview of the articles contained.

The network structure of Hungarian business clusters

This article investigates the attitudes to inter-firm co-operation in Hungary by analysing a special group of business networks: the business clusters. Following an overview of cluster policy, a wide range of selfproclaimed business clusters are identified. A small elite of these business networks evolves into successful, sustainable innovative business clusters. However, in the majority of cases, these consortia of interfirm co-operation are not based on a mutually satisfactory model, and as a consequence, many clusters do not survive in the longer term. The paper uses the concepts and models of social network theory in order to explain, why and under what circumstances inter-firm co-operation in clusters enhances the competitiveness of the network as a whole, or alternatively, under what circumstances the cluster remains dependent on Government subsidies. The empirical basis of the study is a thorough internet research about the Hungarian cluster movement; a questionnaire based expert survey among managers of clusters and member companies and a set of in-depth interviews among managers of self-proclaimed clusters. The last chapter analyises the applicability of social network theory in the analysis of business networks and a model involving the value chain is recommended.

Psychosocial variables associated with the co-occurrence of risky sexual behavior with alcohol consumption among university students

This study examined the association of theoretically guided and empirically identified psychosocial variables on the co-occurrence of risky sexual behavior with alcohol consumption among university students. The study utilized event analysis to determine whether risky sex occurred during the same event in which alcohol was consumed. Relevant conceptualizations included alcohol disinhibition, self-efficacy, and social network theories. Predictor variables included negative condom attitudes, general risk taking, drinking motives, mistrust, social group membership, and gender. Factor analysis was employed to identify dimensions of drinking motives. Measured risky sex behaviors were (a) sex without a condom, (b) sex with people not known very well, (c) sex with injecting drug users (IDUs), (d) sex with people without knowing whether they had a STD, and (e) sex with using drugs. A purposive sample was used and included 222 male and female students recruited from a major urban university. Chi-square analysis was used to determine whether participants were more likely to engage in risky sex behavior in different alcohol use contexts. These contexts were only when drinking, only when not drinking, and when drinking or not. The chi-square findings did not support the hypothesis that university students who use alcohol with sex will engage in riskier sex. These results added to the literature by extending other similar findings to a university student sample. For each of the observed risky sex behaviors, discriminant analysis methodology was used to determine whether the predictor variables would differentiate the drinking contexts, or whether the behavior occurred. Results from discriminant analyses indicated that sex with people not known very well was the only behavior for which there were significant discriminant functions. Gender and enhancement drinking motives were important constructs in the classification model. Limitations of the study and implications for future research, social work practice and policy are discussed. ^

Insulin-like growth factor-I gene expression as a growth indicator in Nile tilapia Oreochromis niloticus L.

Somatic growth in fishes is regulated by a variety of hormones. A central step in this hormonal network is the growth hormone-insulin-like growth factor-I (IGF-I) axis. Studies conducted evaluated the relationship of hepatic IGF-I (hIGF-1) mRNA with growth as affected by feeding regimes (satiation or restricted level; daily or alternate-day feeding), temperatures (high, ambient, low) and by social stress. To develop a cellular means for the quantification of hIGF-I mRNA levels in O. niloticus, hIGF-I cDNA was isolated and cloned. The partial sequence of IGF-I cDNA encodes for signal peptide, mature protein and a portion of the E-domain. A sensitive TaqMan quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed based on the mature IGF-I. Using the developed qRT-PCR assay a significant positive correlation was observed between hIGF-I mRNA levels and growth rate of fish reared under different feeding regimes (r = 0.64) and temperature conditions (r = 0.64). On the dynamics of hIGF-I gene expression in response to elevated temperature, hIGF-I mRNA levels were significantly elevated after at least 2 days of exposure to warm temperature. This validates the concept that hIGF-I gene expressions are sufficiently sensitive to be used as a rapid growth rate indicator for O. niloticus. The hIGF-I levels have a significant positive correlation with specific growth rate (length; r = 0.92), and with condition factor (r = 0.55). On the effect of social stress, differential alterations in growth rates between the dominant and subordinates were observed which was attributed more to behavioral changes as transduced by physiological regulators. The fish's relative position in the social hierarchy was consistently reflected in the levels of hIGF-I mRNA and the eye color pattern. Subordination depressed hIGF-I levels while dominance stimulated it. These findings have shown that hGF-I level remained positively correlated to growth rate as affected by feeding regime, temperature and social stress. This suggests that hIGF-I plays a key role in controlling growth in O. niloticus and indicates that IGF-I mRNA quantification could prove useful for the rapid assessment of growth rate in this species of fish.