Proteomics analysis of human endothelial cell line EA.hy926 after exposure to GSM 900 radiation

The human endothelial cell line EA.hy926 was exposed to mobile phone radiation and the effect on protein expression was examined using two-dimensional electrophoresis (2-DE). Up to 38 various proteins have statistically significantly altered their expression levels following the irradiation. Four proteins were identified with matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). Two of the affected proteins were determined to be isoforms of cytoskeletal vimentin. This finding supports our earlier presented working hypothesis which indicated that the mobile phone radiation might affect the cytoskeleton and might have an effect on the physiological functions that are regulated by the cytoskeleton.     

Mobile phone radiation causes changes in gene and protein expression in human endothelial cell lines and the response seems to be genome- and proteome-dependent

We have examined in vitro cell response to mobile phone radiation (900 MHz GSM signal) using two variants of human endothelial cell line: EA.hy926 and EA.hy926v1. Gene expression changes were examined in three experiments using cDNA Expression Arrays and protein expression changes were examined in ten experiments using 2-DE and PDQuest software. Obtained results show that gene and protein expression were altered, in both examined cell lines, in response to one hour mobile phone radiation exposure at an average specific absorption rate of 2.8 W/kg. However, the same genes and proteins were differently affected by the exposure in each of the cell lines. This suggests that the cell response to mobile phone radiation might be genome- and proteome-dependent. Therefore, it is likely that different types of cells and from different species might respond differently to mobile phone radiation or might have different sensitivity to this weak stimulus. Our findings might also explain, at least in part, the origin of discrepancies in replication studies between different laboratories.     

Mobile phones, heat shock proteins and cancer

There are several reports which indicate that electromagnetic radiation (such as from mobile phones) at non-thermal levels may elicit a biological effect in target cells or tissues. Whether or not these biological effects lead to adverse health effects, including cancer, is unclear. To date there is limited scientific evidence of health issues, and no mechanism by which mobile phone radiation could influence cancer development. In this paper, we develop a theoretical mechanism by which radiofrequency radiation from mobile phones could induce cancer, via the chronic activation of the heat shock response. Upregulation of heat shock proteins (Hsps) is a normal defence response to a cellular stress. However, chronic expression of Hsps is known to induce or promote oncogenesis, metastasis and/or resistance to anticancer drugs. We propose that repeated exposure to mobile phone radiation acts as a repetitive stress leading to continuous expression of Hsps in exposed cells and tissues, which in turn affects their normal regulation, and cancer results. This hypothesis provides the possibility of a direct association between mobile phone use and cancer, and thus provides an important focus for future experimentation.     

Psychophysiological indicators for children using mobile phones. Communication 1. Current state of the problem

An overview of the epidemiological and experimental evidence for exposure of humans and animals to electromagnetic radiation produced by mobile phones is provided. The effects of mobile phone radiation on the child's body are considered in detail. It has been shown that the children's organism is more sensitive to this kind of exposure than the adult one.     

High-throughput analytical gel filtration screening of integral membrane proteins for structural studies

Background

Structural studies of integral membrane proteins (IMPs) are often hampered by difficulties in producing stable homogenous samples for crystallization. To overcome this hurdle it has become common practice to screen large numbers of target proteins to find suitable candidates for crystallization. For such an approach to be effective, an efficient screening strategy is imperative. To this end, strategies have been developed that involve the use of green fluorescent protein (GFP) fusion constructs. However, these approaches suffer from two drawbacks; proteins with a translocated C-terminus cannot be tested and scale-up from analytical to preparative purification is often non-trivial and may require re-cloning.

Methods

Here we present a screening approach that prioritizes IMP targets based on three criteria: expression level, detergent solubilization yield and homogeneity as determined by high-throughput small-scale immobilized metal affinity chromatography (IMAC) and automated size-exclusion chromatography (SEC).

Results

To validate the strategy, we screened 48 prokaryotic IMPs in two different vectors and two Escherichia coli strains. A set of 11 proteins passed all preset quality control checkpoints and was subjected to crystallization trials. Four of these crystallized directly in initial sparse matrix screens, highlighting the robustness of the strategy.

Conclusions

We have developed a rapid and cost efficient screening strategy that can be used for all IMPs regardless of topology. The analytical steps have been designed to be a good mimic of preparative purification, which greatly facilitates scale-up.

General significance

The screening approach presented here is intended and expected to help drive forward structural biology of membrane proteins.     

Non-thermal activation of the hsp27/p38MAPK stress pathway by mobile phone radiation in human endothelial cells: molecular mechanism for cancer- and blood-brain barrier-related effects

Abstract We have examined whether non-thermal exposures of cultures of the human endothelial cell line EA.hy926 to 900 MHz GSM mobile phone microwave radiation could activate stress response. Results obtained demonstrate that 1-hour non-thermal exposure of EA.hy926 cells changes the phosphorylation status of numerous, yet largely unidentified, proteins. One of the affected proteins was identified as heat shock protein-27 (hsp27). Mobile phone exposure caused a transient increase in phosphorylation of hsp27, an effect which was prevented by SB203580, a specific inhibitor of p38 mitogen-activated protein kinase (p38MAPK). Also, mobile phone exposure caused transient changes in the protein expression levels of hsp27 and p38MAPK. All these changes were non-thermal effects because, as determined using temperature probes, irradiation did not alter the temperature of cell cultures, which remained throughout the irradiation period at 37 ± 0.3 °C. Changes in the overall pattern of protein phosphorylation suggest that mobile phone radiation activates a variety of cellular signal transduction pathways, among them the hsp27/p38MAPK stress response pathway. Based on the known functions of hsp27, we put forward the hypothesis that mobile phone radiation-induced activation of hsp27 may (i) facilitate the development of brain cancer by inhibiting the cytochrome c/caspase-3 apoptotic pathway and (ii) cause an increase in blood-brain barrier permeability through stabilization of endothelial cell stress fibers. We postulate that these events, when occurring repeatedly over a long period of time, might become a health hazard because of the possible accumulation of brain tissue damage. Furthermore, our hypothesis suggests that other brain damaging factors may co-participate in mobile phone radiation-induced effects.     

A numerical and experimental comparison of human head phantoms for compliance testing of mobile telephone equipment

A new human head phantom has been proposed by CENELEC/IEEE, based on a large scale anthropometric survey. This phantom is compared to a homogeneous Generic Head Phantom and three high resolution anatomical head models with respect to specific absorption rate (SAR) assessment. The head phantoms are exposed to the radiation of a generic mobile phone (GMP) with different antenna types and a commercial mobile phone. The phones are placed in the standardized testing positions and operate at 900 and 1800 MHz. The average peak SAR is evaluated using both experimental (DASY3 near field scanner) and numerical (FDTD simulations) techniques. The numerical and experimental results compare well and confirm that the applied SAR assessment methods constitute a conservative approach.     

Structural and kinetic effects of mobile phone microwaves on acetylcholinesterase activity

The present study provides evidence that "in vitro" simple exposure of an aqueous solution of electric eel acetylcholinesterase (EeAChE; EC 3.1.1.7.) to cellular phone emission alters its enzymatic activity. This paper demonstrates, by combining different experimental techniques, that radio frequency (RF) radiations irreversibly affect the structural and biochemical characteristics of an important CNS enzyme. These results were obtained by using a commercial cellular phone to reproduce the reality of the human exposition. This experimental procedure provided surprising effects collected practically without experimental errors because they were obtained comparing native and irradiated sample of the same enzyme solution. Although these results cannot be used to conclude whether exposure to RF during the use of cellular phone can lead to any hazardous health effect, they may be a significant first step towards further verification of these effects on other "ex vivo" or "in vivo" biological systems.     

Comparison of cytotoxic and genotoxic effects of plutonium-239 alpha particles and mobile phone GSM 900 radiation in the Allium cepa test

The goal of this study was to compare the cytotoxic and genotoxic effects of plutonium-239 alpha particles and GSM 900 modulated mobile phone (model Sony Ericsson K550i) radiation in the Allium cepa test. Three groups of bulbs were exposed to mobile phone radiation during 0 (sham), 3 and 9 h. A positive control group was treated during 20 min with plutonium-239 alpha-radiation. Mitotic abnormalities, chromosome aberrations, micronuclei and mitotic index were analyzed. Exposure to alpha-radiation from plutonium-239 and exposure to modulated radiation from mobile phone during 3 and 9 h significantly increased the mitotic index. GSM 900 mobile phone radiation as well as alpha-radiation from plutonium-239 induced both clastogenic and aneugenic effects. However, the aneugenic activity of mobile phone radiation was more pronounced. After 9 h of exposure to mobile phone radiation, polyploid cells, three-groups metaphases, amitoses and some unspecified abnormalities were detected, which were not registered in the other experimental groups. Importantly, GSM 900 mobile phone radiation increased the mitotic index, the frequency of mitotic and chromosome abnormalities, and the micronucleus frequency in a time-dependent manner. Due to its sensitivity, the A. cepa test can be recommended as a useful cytogenetic assay to assess cytotoxic and genotoxic effects of radiofrequency electromagnetic fields.     

Early phase drug discovery: Cheminformatics and computational techniques in identifying lead series

Early drug discovery processes rely on hit finding procedures followed by extensive experimental confirmation in order to select high priority hit series which then undergo further scrutiny in hit-to-lead studies. The experimental cost and the risk associated with poor selection of lead series can be greatly reduced by the use of many different computational and cheminformatic techniques to sort and prioritize compounds. We describe the steps in typical hit identification and hit-to-lead programs and then describe how cheminformatic analysis assists this process. In particular, scaffold analysis, clustering and property calculations assist in the design of high-throughput screening libraries, the early analysis of hits and then organizing compounds into series for their progression from hits to leads. Additionally, these computational tools can be used in virtual screening to design hit-finding libraries and as procedures to help with early SAR exploration.     

Testing the effectiveness of small radiation shields for mobile phones

Nine small radiation shields made to adhere to the case of mobile phones were tested at 914 and 1880 MHz. Five popular products were tested because advertisements typically claim they are up to 99% effective in blocking radio frequency (RF) radiation emitted from mobile phones. Also, four other conceptually unusual products were tested because advertisements typically claim they emit oscillations that counteract the RF radiation from mobile phones. Each shield was tested on the same mobile phone, and measurements were made to compare the absorption of RF energy in the head with and without each shield attached to the phone. The phone was positioned against a head model, and an automated measurement process was used to determine specific absorption rate (SAR) in the same way it is used at Motorola to test the compliance of mobile phones with respect to human exposure limits. The location of the peak SAR was not observed to change with any of the shields attached to the phone, and the 1 g, peak spatial average SAR did not change by any statistically significant amount. These results indicate the small shields are ineffective in reducing the exposure of the head to RF energy emitted by a mobile phone.     

Analysis of proteome response to the mobile phone radiation in two types of human primary endothelial cells

Background  

Use of mobile phones has widely increased over the past decade. However, in spite of the extensive research, the question of potential health effects of the mobile phone radiation remains unanswered. We have earlier proposed, and applied, proteomics as a tool to study biological effects of the mobile phone radiation, using as a model human endothelial cell line EA.hy926. Exposure of EA.hy926 cells to 900 MHz GSM radiation has caused statistically significant changes in expression of numerous proteins. However, exposure of EA.hy926 cells to 1800 MHz GSM signal had only very small effect on cell proteome, as compared with 900 MHz GSM exposure. In the present study, using as model human primary endothelial cells, we have examined whether exposure to 1800 MHz GSM mobile phone radiation can affect cell proteome.     

Combinatorial approaches to affinity chromatography

A new armoury of protein purification tools is required to support rapid advances in high-throughput genomics and proteomics, which are predicted to lead to the discovery, isolation, characterisation and manufacture of a number of new biopharmaceutical proteins. Computer-aided molecular design, combinatorial (bio)chemistry and high-throughput screening techniques are now being exploited to identify highly selective ligands for use in the purification of these proteins by affinity chromatography.     

High-throughput expression,purification, and characterization of recombinant Caenorhabditis elegans proteins

Modern proteomics approaches include techniques to examine the expression, localization, modifications, and complex formation of proteins in cells. In order to address issues of protein function in vitro using classical biochemical and biophysical approaches, high-throughput methods of cloning the appropriate reading frames, and expressing and purifying proteins efficiently are an important goal of modern proteomics approaches. This process becomes more difficult as functional proteomics efforts focus on the proteins from higher organisms, since issues of correctly identifying intron-exon boundaries and efficiently expressing and solubilizing the (often) multi-domain proteins from higher eukaryotes are challenging. Recently, 12,000 open-reading-frame (ORF) sequences from Caenorhabditis elegans have become available for functional proteomics studies [Nat. Gen. 34 (2003) 35]. We have implemented a high-throughput screening procedure to express, purify, and analyze by mass spectrometry hexa-histidine-tagged C. elegans ORFs in Escherichia coli using metal affinity ZipTips. We find that over 65% of the expressed proteins are of the correct mass as analyzed by matrix-assisted laser desorption MS. Many of the remaining proteins indicated to be "incorrect" can be explained by high-throughput cloning or genome database annotation errors. This provides a general understanding of the expected error rates in such high-throughput cloning projects. The ZipTip purified proteins can be further analyzed under both native and denaturing conditions for functional proteomics efforts.     

Simplifying a life cycle assessment of a mobile phone

Purpose

The possibilities for full life cycle assessment (LCA) of new Information and Communication Technology (ICT) products are often limited, so simplification approaches are needed. The aim of this paper is to investigate possible simplifications in LCA of a mobile phone and to use the results to discuss the possibilities of LCA simplifications for ICT products in a broader sense. Another aim is to identify processes and data that are sensitive to different methodological choices and assumptions related to the environmental impacts of a mobile phone.

Methods

Different approaches to a reference LCA of a mobile phone was tested: (1) excluding environmental impact categories, (2) excluding life cycle stages/processes, (3) using secondary process data from generic databases, (4) using input-output data and (5) using a simple linear relationship between mass and embodied emissions.

Results and discussion

It was not possible to identify one or a few impact categories representative of all others. If several impact categories would be excluded, information would be lost. A precautionary approach of not excluding impact categories is therefore recommended since impacts from the different life cycle stages vary between impact categories. Regarding use of secondary data for an ICT product similar to that studied here, we recommend prioritising collection of primary (specific) data on energy use during production and use, key component data (primarily integrated circuits) and process-specific data regarding raw material acquisition of specific metals (e.g. gold) and air transport. If secondary data are used for important processes, the scaling is crucial. The use of input-output data can be a considerable simplification and is probably best used to avoid data gaps when more specific data are lacking.

Conclusions

Further studies are needed to provide for simplified LCAs for ICT products. In particular, the end-of-life treatment stage need to be further addressed, as it could not be investigated here for all simplifications due to data gaps.     

Ionizing and non-ionizing radiation (comparative risk estimations)

The population has widely used mobile communication for already more than 15 years. It is important to note that the use of mobile communication has sharply changed the conditions of daily exposure of the population to EME We expose our brain daily for the first time in the entire civilization. The mobile phone is an open and uncontrollable source of electromagnetic radiation. The comparative risk estimation for the population of ionizing and non-ionizing radiation was carried out taking into account the real conditions of influence. Comparison of risks for the population of ionizing and non-ionizing radiation leads us to a conclusion that EMF RF exposure in conditions of wide use of mobile communication is potentially more harmful than ionizing radiation influence.     

‘Do you have a mobile?’ Mobile phone practices and the refashioning of social relationships in Port Vila Town

Mobile phones have quickly become an important part of young people's social relationships in Port Vila Vanuatu. In particular, young people embrace the new technology's capacity to broaden the breadth of their sociality. They use the mobile phone to facilitate private and secretive communication, engage in unsanctioned relationships, pre‐marital sexual relationships, and also multiple concurrent intimate relationships. Literature on mobile phone use often either takes the approach that mobile phone technology becomes purely incorporated into pre‐existing social practices, or that it dramatically reshapes social ontologies. The present article argues for an alternative view, one that takes into account the nuances between these two analytical poles. This article suggests that young people use the mobile phone in practices informed by previous models of social relationships, yet the specific materiality of the mobile phone technology is influencing the direction in which models of social relationships are changing. In demonstrating this point, I pay particular attention to two material aspects of the mobile phone technology ‐ the mobile phone as a repository of a particular kind of information ‐ ‘evidence’, and the capacity of the mobile phone to ‘disconnect’ people from their relationships by switching off the mobile. This article argues that these practices are influencing the emergence of a radically altered kinship and gender landscape in the urban context.     

On the Use of Human Mobility Proxies for Modeling Epidemics

Human mobility is a key component of large-scale spatial-transmission models of infectious diseases. Correctly modeling and quantifying human mobility is critical for improving epidemic control, but may be hindered by data incompleteness or unavailability. Here we explore the opportunity of using proxies for individual mobility to describe commuting flows and predict the diffusion of an influenza-like-illness epidemic. We consider three European countries and the corresponding commuting networks at different resolution scales, obtained from (i) official census surveys, (ii) proxy mobility data extracted from mobile phone call records, and (iii) the radiation model calibrated with census data. Metapopulation models defined on these countries and integrating the different mobility layers are compared in terms of epidemic observables. We show that commuting networks from mobile phone data capture the empirical commuting patterns well, accounting for more than 87% of the total fluxes. The distributions of commuting fluxes per link from mobile phones and census sources are similar and highly correlated, however a systematic overestimation of commuting traffic in the mobile phone data is observed. This leads to epidemics that spread faster than on census commuting networks, once the mobile phone commuting network is considered in the epidemic model, however preserving to a high degree the order of infection of newly affected locations. Proxies'' calibration affects the arrival times'' agreement across different models, and the observed topological and traffic discrepancies among mobility sources alter the resulting epidemic invasion patterns. Results also suggest that proxies perform differently in approximating commuting patterns for disease spread at different resolution scales, with the radiation model showing higher accuracy than mobile phone data when the seed is central in the network, the opposite being observed for peripheral locations. Proxies should therefore be chosen in light of the desired accuracy for the epidemic situation under study.