EHV-1 is a double-stranded DNA virus whereas EAV is a positive sense, single-stranded RNA virus. Therefore, genetically, they are very different from one another. However, both these viruses are endotheliotropic and thus, infect and replicates in equine endothelial cells resulting in vasculitis. Vasculitis is central to the pathogenesis of these two viruses. Thus, the main objective of this thesis was to investigate the inflammatory and innate immune responses of EECs that contribute towards the development of vasculitis following infection with EHV-1 and EAV in-vitro. Since proinflammatory cytokines and chemokines produced by endothelial cells play a significant role in the development of vasculitis, we investigated their gene expression as well as secretion. Results from this study showed that the proinflammatory response of EECs induced by EAV is relatively less when compared with the corresponding results from EHV-1 infected EECs. Furthermore, EAV elicits a lower type I interferon response in EECs when compared with EHV-1. Further investigations revealed an active role played by TLR 3 in inducing the proinflammatory response in EHV-1 infected EECs during the first 6 hours of infection but not in EAV infected EECs. Analyzing the whole transcriptome of EHV-1 and EAV infected EECs revealed a complex pattern of gene regulation and cellular pathways related to cellular immune, inflammatory and apoptotic responses. Finally, we investigated host genetic factors associated with EHV-1 induced myeloencephalopathy but found no evidence for a recessive allele influencing the development of EHM following EHV-1 infection for any genetic locus was identified. However, more complex host-pathogen interactions are possible.

O Herpesvírus equino tipo -1 (EHV-1) pertence ao gênero Varicellovírus da subfamília Alphaherpesvirinae pertencente à Família Herpesviridae. É um vírus envelopado, de DNA linear fita dupla, composto por 76 genes distintos. O EHV-1 é responsável por grandes prejuízos econômicos na equinocultura mundial. Responsável por doença neonatal fatal, mieloencefalopatia, rinopneumonite e abortamento, encontra-se amplamente distribuído pela população equina do território nacional. O objetivo do presente estudo foi o de padronizar uma reação em cadeia pela polimerase (PCR) para detecção do EHV-1 em tecidos incluídos em parafina a fim de permitir estudos retrospectivos em arquivos de amostras histopatológicas. Assim, foram inoculados experimentalmente 12 camundongos com 21 dias de idade da linhagem CH3/Rockfeller com três diferentes isolados de EHV-1, dois provenientes da Argentina e um do Brasil. Esses animais foram observados por quatro dias e, após sacrifício por sobre dose de uma associação de ketamina e xilazina, foram submetidos à necropsia e colhidos o pulmão e sistema nervoso central (SNC). Os órgãos colhidos foram divididos em duas partes aproximadamente iguais: uma mantida a -20ºC até processamento e a outra fixada em formalina 10% tamponada e posteriormente incluída em parafina. A extração foi realizada com nove fragmentos contínuos de 4µm cada, a partir do protocolo de extração com proteinase K/ fenol/ clorofórmio. Foi realizada avaliação da sensibilidade analítica da PCR com oito diluições na base 10 para os três isolados utilizados. A amplificação do DNA viral foi realizada utilizando primers direcionados para a ORF64. A fim de descartar a eventual presença de inibidores da reação de PCR e assegurar a adequada extração de DNA, foram incluídos primers direcionados para o gene da beta-actina. A PCR mostrou-se capaz de amplificar DNA viral alvo numa diluição de até 10-5, sendo positiva entre 10-1 a 10-2 DICT50/25µL. Com a PCR padronizada, foi possível detectar o DNA do EHV-1 em: a) 100% (12/12) das amostras de pulmão congeladas e 100% (12/12) das amostras de pulmão incluídas em parafina; b) em 91% (11/12) das amostras de SNC congeladas e 41% (5/12) das amostras de SNC incluídas em parafina. A aplicação da PCR padronizada em uma coleção de amostras incluídas em parafina do Laboratório de Anatomia Patológica do IB/SP, colhidas de cinco casos de abortamento em equinos, revelou que o DNA do EHV-1 foi detectado em: a) um caso em que originalmente foi possível isolar o EHV-1; b) em 4/4 amostras que revelaram-se originalmente negativas. Com base nos resultados obtidos, foi possível concluir que a PCR padronizada teve bom desempenho na detecção de DNA viral em amostras incluídas em parafina de animais experimentalmente infectados e, provavelmente, uma sensibilidade diagnóstica mais elevada que os métodos utilizados para o diagnóstico do EHV-1 na coleção de amostras de equino testada. / The equine herpesvirus type 1 (EHV-1) belongs to Varicellovírus genus, Alphaherpesvirinae subfamíly of the Herpesviridae Famíly. It is an enveloped virus, double stranded linear DNA, composed of 76 distinct genes. The EHV-1 is responsible for great losses in horsebread world. Responsible for neonatal death, mieloencephalopaty, rinopneumonite and abortion, it is widely distributed into brasilian equine population. The purpose of this study was to standardize a polymerase chain reaction (PCR) for EHV-1detection in paraffin- embedded tissues allowing retrospective studies based on the collection histopatological samples. Thus, 12 mice (CH3/Rockfeller) with 21 days of age were inoculated with 3 different isolates of EHV-1, 2 from Argentina and one from Brazil. These mice were observed for 4 days and, after sacrifice by overdose of a combination of ketamine and xylazina, it were subjected to necropsy and collected the lung and central nervous system (SNC). The collected tissues were divided into 2 approximately equal parts: 1one stored at -20ºC until processing, and another set at 10% buffering formalin and later paraffin-embedded. The extraction was performed with continuous 9 fragments of 4µm each, using the extration protocol with proteinase K/ fenol/ clorofórmio. The assessment of analytical sensitivity of PCR were determined using 8 dilutions for all 3 virus isolates. The viral DNA amplification was performed using primers targeted to ORF64. In order to rule out the possible presence of PCR inhibitors and to ensure adequate extraction of DNA, primers directed to the gene for beta-actin were included It was possible to amplify viral DNA until 10-5 dilution, corresponding to 10-1 to 10-2 DICT50/25µL. With the standardized PCR, it was possible to detect the EHV-1 DNA in: a) 100% (12/12) of lung frozen sample and 100% (12/12) of the paraffin-embedded lung; b) 91% (11/12) of the frozen CNS and 41% (5/12) CNS paraffin-embedded. Moreover, the standardized PCR was tested in a collection of paraffin-embedded specimens from Pathological Anatomy Laboratory od Biological Institute Sao Paulo State, taken 5 cases of the abortion in horses. It were possible to detect EHV-1 DNA in: a) 1 sample from a case in that originally was possible to isolate the EHV-1, b) 4/4 sample originally negative diagnosed. Based on these results, it is possible to conclude that the standardized PCR performed well for detection viral DNA in paraffin-embedded tissues of experimentally infected animals, and probably a higher diagnostic sensitivity than the methods used for diagnosis of EHV-1 in the collection samples tissues tested for equine.

Equine herpesvirus type-1(EHV-1) is a complex virus known for inducing various forms of disease in horses. In recent years, the number of cases of neurological disease caused by this virus has increased. While there are a number of possible sources for this recent surge, this project set out to determine if a genotypic shift in the latent population of the virus in favor of the neuropathogenic form of EHV-1 is the basis for the recent increase in frequency of EHV-1 neurologic disease. To ascertain if such a shift has in fact occurred, 450 EHV-1 isolates were obtained from fetal tissues resulting from single, sporadic cases of abortion in Thoroughbred broodmares in central Kentucky. Furthermore, the isolates utilized were from different decades (1951-2006) to determine if the genotypic shift was time-related. The isolates were propagated in cell culture, purified and the viral DNA isolated. Real-time allelic discrimination PCR analysis was performed on the DNA samples to identify the genotype of EHV-1. Statistical analysis of the PCR data indicates that the latent mutant population does appear to be increasing. Therefore, the recent increase in the number of outbreaks of EHV-1 neurological disease will most likely continue unless measures are devised to curtail further spread of the pathogen.

O Herpesvírus equino tipo -1 (EHV-1) pertence ao gênero Varicellovírus da subfamília Alphaherpesvirinae pertencente à Família Herpesviridae. É um vírus envelopado, de DNA linear fita dupla, composto por 76 genes distintos. O EHV-1 é responsável por grandes prejuízos econômicos na equinocultura mundial. Responsável por doença neonatal fatal, mieloencefalopatia, rinopneumonite e abortamento, encontra-se amplamente distribuído pela população equina do território nacional. O objetivo do presente estudo foi o de padronizar uma reação em cadeia pela polimerase (PCR) para detecção do EHV-1 em tecidos incluídos em parafina a fim de permitir estudos retrospectivos em arquivos de amostras histopatológicas. Assim, foram inoculados experimentalmente 12 camundongos com 21 dias de idade da linhagem CH3/Rockfeller com três diferentes isolados de EHV-1, dois provenientes da Argentina e um do Brasil. Esses animais foram observados por quatro dias e, após sacrifício por sobre dose de uma associação de ketamina e xilazina, foram submetidos à necropsia e colhidos o pulmão e sistema nervoso central (SNC). Os órgãos colhidos foram divididos em duas partes aproximadamente iguais: uma mantida a -20ºC até processamento e a outra fixada em formalina 10% tamponada e posteriormente incluída em parafina. A extração foi realizada com nove fragmentos contínuos de 4µm cada, a partir do protocolo de extração com proteinase K/ fenol/ clorofórmio. Foi realizada avaliação da sensibilidade analítica da PCR com oito diluições na base 10 para os três isolados utilizados. A amplificação do DNA viral foi realizada utilizando primers direcionados para a ORF64. A fim de descartar a eventual presença de inibidores da reação de PCR e assegurar a adequada extração de DNA, foram incluídos primers direcionados para o gene da beta-actina. A PCR mostrou-se capaz de amplificar DNA viral alvo numa diluição de até 10-5, sendo positiva entre 10-1 a 10-2 DICT50/25µL. Com a PCR padronizada, foi possível detectar o DNA do EHV-1 em: a) 100% (12/12) das amostras de pulmão congeladas e 100% (12/12) das amostras de pulmão incluídas em parafina; b) em 91% (11/12) das amostras de SNC congeladas e 41% (5/12) das amostras de SNC incluídas em parafina. A aplicação da PCR padronizada em uma coleção de amostras incluídas em parafina do Laboratório de Anatomia Patológica do IB/SP, colhidas de cinco casos de abortamento em equinos, revelou que o DNA do EHV-1 foi detectado em: a) um caso em que originalmente foi possível isolar o EHV-1; b) em 4/4 amostras que revelaram-se originalmente negativas. Com base nos resultados obtidos, foi possível concluir que a PCR padronizada teve bom desempenho na detecção de DNA viral em amostras incluídas em parafina de animais experimentalmente infectados e, provavelmente, uma sensibilidade diagnóstica mais elevada que os métodos utilizados para o diagnóstico do EHV-1 na coleção de amostras de equino testada. / The equine herpesvirus type 1 (EHV-1) belongs to Varicellovírus genus, Alphaherpesvirinae subfamíly of the Herpesviridae Famíly. It is an enveloped virus, double stranded linear DNA, composed of 76 distinct genes. The EHV-1 is responsible for great losses in horsebread world. Responsible for neonatal death, mieloencephalopaty, rinopneumonite and abortion, it is widely distributed into brasilian equine population. The purpose of this study was to standardize a polymerase chain reaction (PCR) for EHV-1detection in paraffin- embedded tissues allowing retrospective studies based on the collection histopatological samples. Thus, 12 mice (CH3/Rockfeller) with 21 days of age were inoculated with 3 different isolates of EHV-1, 2 from Argentina and one from Brazil. These mice were observed for 4 days and, after sacrifice by overdose of a combination of ketamine and xylazina, it were subjected to necropsy and collected the lung and central nervous system (SNC). The collected tissues were divided into 2 approximately equal parts: 1one stored at -20ºC until processing, and another set at 10% buffering formalin and later paraffin-embedded. The extraction was performed with continuous 9 fragments of 4µm each, using the extration protocol with proteinase K/ fenol/ clorofórmio. The assessment of analytical sensitivity of PCR were determined using 8 dilutions for all 3 virus isolates. The viral DNA amplification was performed using primers targeted to ORF64. In order to rule out the possible presence of PCR inhibitors and to ensure adequate extraction of DNA, primers directed to the gene for beta-actin were included It was possible to amplify viral DNA until 10-5 dilution, corresponding to 10-1 to 10-2 DICT50/25µL. With the standardized PCR, it was possible to detect the EHV-1 DNA in: a) 100% (12/12) of lung frozen sample and 100% (12/12) of the paraffin-embedded lung; b) 91% (11/12) of the frozen CNS and 41% (5/12) CNS paraffin-embedded. Moreover, the standardized PCR was tested in a collection of paraffin-embedded specimens from Pathological Anatomy Laboratory od Biological Institute Sao Paulo State, taken 5 cases of the abortion in horses. It were possible to detect EHV-1 DNA in: a) 1 sample from a case in that originally was possible to isolate the EHV-1, b) 4/4 sample originally negative diagnosed. Based on these results, it is possible to conclude that the standardized PCR performed well for detection viral DNA in paraffin-embedded tissues of experimentally infected animals, and probably a higher diagnostic sensitivity than the methods used for diagnosis of EHV-1 in the collection samples tissues tested for equine.

High voltage (EHV and UHV) transmission facilitates transfer of large amount of power over long distances. However, due to the inherent geometry, the line and substation hardware of EHV and UHV class generate high electric fields, which results in local ionisation of air called corona discharges. Apart from producing audible noise in the form of frying or hissing sound, corona produces significant electromagnetic interferences in the radio range. The limit for this corona generated Radio Interference (RI) has been stipulated by international standards, which are strictly to be followed. In line and substation hardware, corona control rings are generally employed to limit or avoid corona. Standard dimensions of corona rings are not available for EHV and UHV class. In most of the cases, their design is based on either a trial and error method or based on empirical extrapolation. Only in certain specific cases, the dimensioning of the rings is carried out using electric field calculations. In any of these approaches, the unavoidable surface abrasions, which can lead to corona, are not considered. There are also efforts to account for nominal surface irregularity by using a surface roughness factor, which is highly heuristic. In order to address this practically relevant problem, the present work was taken up. The intended exercise requires accurate field computation and a suitable criterion for checking corona onset. For the first part, the Surface Charge Simulation Method is adopted with newly proposed sub-modelling technique. The surface of the toroid is discretised into curvilinear patches with linear approximation for the surface charge density. Owing to its high accuracy, Galerkin’s method of moments formulation is employed. The problem of singularity encountered in the numerical approach is handled using a method based on Duffy’s transformation. The developed codes have also been validated with standard geometries. After a survey of relevant literature the ‘Critical Avalanche Criteria’ is chosen for its simplicity and applicability to the problem. Through a detailed simulation, the effect of avalanche space charge in reducing the corona onset voltage is found to be around 1.5% and hence it is not considered further. For utilities not interested in a detailed calculation procedure for dimensioning of corona rings, design curves are developed for circular corona rings of both 400 kV and 765 kV class with surface roughness factor in the range 0.8 – 1. In the second part of the work, a methodology for dimensioning is developed wherein the inevitable surface abrasion in the form of minute protrusions can be accounted. It is first shown that even though considerable field intensification occurs at the protrusions, such localised modification need not lead to corona. It is shown that by varying the minor radius of the corona ring, it is possible to get a design where the prescribed surface abrasion does not lead to corona onset. In summary, the present work has successfully developed a reliable methodology for the design of corona rings with prescribed surface abrasions. It involved development of an efficient field computation technique for handling minute surface protrusions and use of appropriate criteria for assessing corona inception. It has also provided design curves for EHV and UHV class corona rings with surface roughness factor specified in the range 0.8 – 1.0.

I mitt arbete ämnar jag besvara frågan “Hur uttrycks stora fordonstillverkares eco-branding i förhållande till paradigmskiftet mot krav på hållbar utveckling?”. Syftet är att undersöka svårigheterna och balansgången med eco-branding inom fordonsindustrin. Detta görs utifrån en branding-studie av tre stora fordonstillverkares användning av eco-branding, ihop med mina egna erfarenheter av ett liknande designprojekt. Grafisk identitet och visuell kommunikation studeras utifrån el och hybridbilarna BMW i3, Tesla Model S och Toyota Prius. Positionering av företagen inom området kopplas till teori om hållbar utveckling, eco-branding och greenwashing för att diskutera krav på transparens och företagens upplevda legitimitet och trovärdighet. Det framkommer att de olika företagen brandar sig på olika sätt, med olika grad av eco-branding. Något som beror mycket på vilka värderingar och historia företaget och dess varumärke har. Även om det är en balansgång mellan greenwashing och trovärdighet, så kan alla ses ha olika styrkor; samt svagheter att förbättra för ökad transparens eller trovärdighet. En slutsats är dock att eco-branding börjar bli mer trovärdig och att greenwashing-trenden generellt börjat mattas av. Gemensamt för alla aktörer är att framtidsvisionen är det centrala i varumärkesbyggandet. Dock har konventionella fordonstillverkare svårare förutsättningar att förhålla sig till paradigmskiftet än nyare aktörer. / In this essay I aim to answer the question “How is the eco-branding of large automotive companies expressed, in relation to the paradigm shift towards demands for sustainability?”. The purpose is to examine the difficulties and balancing of eco-branding within the automotive industry. This is done through a comparative analysis of three large automotive companies and their use of eco-branding, together with my own experience from a similar design project. Corporate identity and visual communication is studied through the electric and hybrid cars BMW i3, Tesla Model S and Toyota Prius. The positioning of the brands within the field is linked to theory on sustainable development, eco-branding and greenwashing; to discuss demands for transparency and the brands perceived legitimacy and credibility. It is revealed that the different companies brand themselves in relatively different ways, with different degrees of eco-branding.This depends on the values and history of the company and it’s brand. Even though there is a balancing between greenwashing and credibility, they all have different strengths, as well as areas for improvement to increase company transparency and authenticity. A conclusion is however that eco-branding is becoming more credible and the trend of greenwashing is generally started to slightly diminish. It is revealed that future vision plays a central part in all of the brands. However conventional companies are shown to have significantly more difficulties in positioning themselves within the paradigmshift compared to newer actors.

Ever increasing demand for electricity, exploitation of large hydro and nuclear power at remote location has led to power evacuation by long EHV/UHV transmission systems. This thesis concentrates on transient analysis of EHV/UHV transmission systems for improved planning and protection. In this thesis, the uncontrolled and controlled switching methods to limit the switching surges during energization of 765kV and 1200k VUHV transmission lines are studied. The switching surge over voltages during the energization of series compensated line are compared with uncompensated line. A Generalized Electromagnetic Transients Program has been developed. The program incorporates specific models for studying the effectiveness of various means for control of switching surge over voltages during UHV transmission line energization and also simulation of various types of faults. Since power grids may adopt next higher UHV transmission level 1200kV, these studies are necessary for insulation coordination as well as transmission line protection relay settings. A new fault detection/location technique is presented for transmission line using synchronized fundamental voltage and current phasors obtained by PMUs at both ends of line. It is adaptive to fault resistance, source impedance variation, line loading and fault incidence angle. An improved Discrete Fourier Transform (DFT) algorithm to estimate and eliminate the decaying dc component in a fault current signal is proposed for computing the phasors. The settings for digital distance relays under different operating conditions are obtained. The relay should operate faster and be more sensitive to various faults under different conditions without loosing selectivity. An accurate faulted transmission line model which considers distributed shunt capacitance has been presented. The relay trip boundaries are obtained considering transmission line model under realistic fault conditions. For different loading conditions ideal relay characteristic has been developed. The obtained trip boundaries can be used for proper settings of practical relay. An adaptive relaying scheme is proposed for EHV/UHV transmission line using unsynchronized/synchronized fundamental voltage and current phasors at both ends of line. For fault location, the redundancy in equations is achieved by using two kinds of Clarke’s components which makes the calculations non-iterative and accurate. An operator for synchronization of the unsynchronized measurements is obtained by considering the distributed parameter line model. The distance to fault is calculated as per the synchronized measurements. Support Vector Machine(SVM) is used for high speed protection of UHV line. The proposed relaying scheme detects the fault and faulted phase effectively within few milli seconds. The current and voltage signals of all phases at the substation are fed to SVM directly at a sampling frequency of 1.0kHZi.e20 samples/cycle . It is possible to detect faulted phase with in 3msec, using the data window of 1/4th cycle. The performance of relaying scheme has been checked with a typical 765kV Indian transmission System which is simulated using the developed EMTP.

The modern day power system is witnessing a tremendous change. There has been a rapid rise in the distributed generation, along with this the deregulation has resulted in a more complex system. The power demand is on a rise, the generation and trans-mission infrastructure hasn't yet adapted to this growing demand. The economic and operational constraints have forced the system to be operated close to its design limits, making the system vulnerable to disturbances and possible grid failure. This makes the study of voltage stability of the system important more than ever. Generally, voltage stability studies are carried on a single phase equivalent system assuming that the system is perfectly balanced. However, the three phase power system is not always in balanced state. There are a number of untransposed lines, single phase and double phase lines. This thesis deals with three phase voltage stability analysis, in particular the voltage stability index known as L-Index. The equivalent single phase analysis for voltage stability fails to work in case of any unbalance in the system or in presence of asymmetrical contingency. Moreover, as the system operators are giving importance to synchrophasor measurements, PMUs are being installed throughout the system. Hence, the three phase voltages can be obtained, making three phase analysis easier. To study the effect of unbalanced system on voltage stability a three phase L-Index based on traditional L-Index has been proposed. The proposed index takes into consideration the unbalance resulting due to untransposed transmission lines and unbalanced loads in the system. This index can handle any unbalance in the system and is much more realistic. To obtain bus voltages during unbalanced operation of the system a three phase decoupled Newton Raphson load ow was used. Reactive power distribution in a system can be altered using generators voltage set-ting, transformers OLTC settings and SVC settings. All these settings are usually in balanced mode i.e. all the phases have the same setting. Based on this reactive power optimization using LP technique on an equivalent single phase system is proposed. This method takes into account generator voltage settings, OLTC settings of transformers and SVC settings. The optimal settings so obtained are applied to corresponding three phase system. The effectiveness of the optimal settings during unbalanced scenario is studied. This method ensures better voltage pro les and decrease in power loss. Case studies of the proposed methods are carried on 12 bus and 24 bus EHV systems of southern Indian grid and a modified IEEE 30 bus system. Both balanced and unbalanced systems are studied and the results are compared.