Oxidação de proteínas por oxigênio singlete: mecanismos de dano, estratégias para detecção e implicações biológicas

Proteins are potential targets for singlet molecular oxygen (¹O2) oxidation. Damages occur only at tryptophan, tyrosine, histidine, methionine, and cysteine residues at physiological pH, generating oxidized compounds such as hydroperoxides. Therefore, it is important to understand the mechanisms by which ¹O2, hydroperoxides and other oxidized products can trigger further damage. The improvement and development of new tools, such as clean sources of ¹O2 and isotopic labeling approaches in association with HPLC/mass spectrometry detection will allow one to elucidate mechanistic features involving ¹O2-mediated protein oxidation.

Estresse oxidativo, lesões no genoma e processos de sinalização no controle do ciclo celular

The generation of reactive oxygen species (ROS) may be both beneficial to cells, performing functions in intracellular signaling and detrimental, modifying cellular biomolecules. ROS can cause DNA damage, such as base damage and strand breaks. Organisms respond to chromosome insults by activation of a complex and hierarchical DNA-damage response pathway. The extent of DNA damages determines cell fate: cell cycle arrest and DNA repair or cell death. The ATM is a central protein in the response to DNA double-strand breaks by acting as a transducer protein. Collected evidences suggest that ATM is also involved in the response to oxidative DNA damage.

The Detector Control Systems for the CMS Resistive Plate Chamber at LHC

The RPC Detector Control System (RCS) is the main subject of this PhD work. The project, involving the Lappeenranta University of Technology, the Warsaw University and INFN of Naples, is aimed to integrate the different subsystems for the RPC detector and its trigger chain in order to develop a common framework to control and monitoring the different parts. In this project, I have been strongly involved during the last three years on the hardware and software development, construction and commissioning as main responsible and coordinator. The CMS Resistive Plate Chambers (RPC) system consists of 912 double-gap chambers at its start-up in middle of 2008. A continuous control and monitoring of the detector, the trigger and all the ancillary sub-systems (high voltages, low voltages, environmental, gas, and cooling), is required to achieve the operational stability and reliability of a so large and complex detector and trigger system. Role of the RPC Detector Control System is to monitor the detector conditions and performance, control and monitor all subsystems related to RPC and their electronics and store all the information in a dedicated database, called Condition DB. Therefore the RPC DCS system has to assure the safe and correct operation of the sub-detectors during all CMS life time (more than 10 year), detect abnormal and harmful situations and take protective and automatic actions to minimize consequential damages. The analysis of the requirements and project challenges, the architecture design and its development as well as the calibration and commissioning phases represent themain tasks of the work developed for this PhD thesis. Different technologies, middleware and solutions has been studied and adopted in the design and development of the different components and a big challenging consisted in the integration of these different parts each other and in the general CMS control system and data acquisition framework. Therefore, the RCS installation and commissioning phase as well as its performance and the first results, obtained during the last three years CMS cosmic runs, will be

Raios-x: fascinação, medo e ciência

This work presents the discovery and the use of x-rays at the end of the XIXth and the beginning of the XXth century. X-rays greatly impacted science and everyday life. Their existence broke the idea that knowledge had reached a limiting step. In general, people regarded x-rays as a marvel of science, but reactions against their use were also found. Several applications were proposed, especially in medicine. However, little or no attention was paid to security measures, leading to health damages and even death. The development of the radiological protection took into account the accidents with the x-rays.

Corporate Governance: Osakeyhtiön johdon velvollisuudet ja vastuu

Tutkimuksen tarkoituksena on selvittää, millainen Corporate Governance -järjestelmä ja sääntely-ympäristö Suomessa on voimassa. Tutkimuksessa tarkastellaan lisäksi osakeyhtiön johdon fidusiaarisia velvollisuuksia ja johdon velvollisuuksien tehosteena olevaa vastuujärjestelmää. Corporate Governance -järjestelmää analysoidaan perinteisestä oikeustieteellisestä näkökulmasta poiketen oikeustaloustieteen metodein. Tutkimuksen teoriapohja perustuu koti- ja ulkomaiseen oikeustaloustieteelliseen kirjallisuuteen, muutoin tutkimuksessa tukeudutaan suurimmilta osin kotimaiseen oikeuskirjallisuuteen ja virallislähteisiin. Suomalainen Corporate Governance -järjestelmä minimioi tehokkaasti transaktiokustannuksia ja tarjoaa osakkeenomistajille tehokkaan, informaation epäsymmetriaa rajoittavan valvontamekanismin. Jos johto toimii velvollisuuksiensa vastaisesti, osakkeenomistajat erottavat johdon, joka ei nauti sen luottamusta tai johto joutuu vastaamaan toimistaan vahingonkorvausvastuun tai rikosoikeudellisen vastuun kautta.

Vesiteiden pato- ja sulkurakenteiden korjaustarpeen määrittäminen Suomen merkittävimmissä vesivoimalaitoksissa

Tämän diplomityön tavoitteena oli kartoittaa suomalaisten vesivoimalaitosten vesiteiden pato – ja sulkurakenteiden tulevia korjaustarpeita seuraavien viiden vuoden aikana. Tutkimuskohteiksi rajattiin yli 1 MW:n ja ennen vuotta 1975 valmistuneet sekä vielä peruskorjaamattomat kohteet. Näillä rajauksilla muodostui 73 vesivoimalaitoksen joukko. Näissä vesivoimalaitoksissa oli yhteensä 133 koneistoa ja 394 erilaista sulkurakennetta. Tutkimusaineisto hankittiin haastattelemalla 30 henkilöä yhteensä 24:stä eri yhtiöstä. Haastateltavat olivat pääasiassa laitosten käyttö- tai kunnossapitopäälliköitä, mutta myös omistajayhtiöiden ylintä johtoa. Strukturoidun haastattelulomakkeen avulla toteutettuna haastattelu oli siten lähes puhdas kyselytutkimus, jonka vastausprosentiksi muodostui 100 %. Haastattelu mahdollisti epäselvien yksityiskohtien selvittämisen ja siten tutkimuksen luotettavuus muodostui suuremmaksi kuin pelkällä kyselylomake - mittauksella. Tulokset osoittivat, että tulevia korjaustarpeita havaittiin noin kolmasosassa tutkimuskohteista. Tyypillisimmät korjaustarpeet ovat vesiteiden betonirakenteisiin tulleissa vaurioissa sekä sulkurakenteiden teknisen ikääntymisen myötä tulevissa tiiviysongelmissa sekä toiminnallisissa vioissa.

Response of cucurbit species to Rotylenchulus reniformis

The state of Rio Grande do Norte is the major melon (Cucumis melo) producer in Brazil and the reniform nematode Rotylenchulus reniformis has become increasingly important due to damages to that crop and the lack of resistant cultivars and effective control measures. Under these circumstances, eight cucurbit genotypes were screened for resistance or tolerance to a population of R. reniformis in naturally infested field soil, under greenhouse conditions. A high reduction in shoot mass was found in all infected genotypes. Watermelon (Citrullus lanatus) cv. Sugar Baby showed the lowest rate of nematode reproduction.

Irtokappaleiden hallinnan nykytila ja kehittäminen Loviisan voimalaitoksella

Ydinvoimateollisuudessa irtokappaleella tarkoitetaan prosessiin kuulumatonta materiaalia, joka ei luonnostaan kuulu prosessiin ja jolla prosessiin jäädessään voi olla haitallinen vaikutus sen toimintaan, komponentteihin tai kemiaan. Irtokappaleet voivat aiheuttaa monenlaisia haittoja ydinvoimalaitoksen turvalliselle ja taloudelliselle käytölle. Vaikka ydinvoimalaitoksen käytössä turvallisuus on aina etusijalla, negatiiviset vaikutukset turvallisuuteen heijastuvat usein myös talouteen. Diplomityössä arvioitiin erilaisia irtokappaleiden aiheuttamia riskejä ydinvoimalan toiminnalle käyttötapahtumia analysoimalla. Erilaisiin tapahtumiin, niiden aiheuttajiin ja vaikutusmekanismeihin tutustumisen jälkeen tutkittiin Loviisan voimalaitoksen käyttämiä toimintatapoja irtokappaleiden hallitsemiseksi. Irtokappaleiden hallintaan pyritään sekä hallinnollisilla että käytännön menetelmillä. Hallinnollisia menetelmiä ovat esimerkiksi ohjeistojärjestelmä ja johtaminen, käytännön menettelytapoja esimerkiksi työtavat ja koulutus. Irtokappaleiden hallinnan menetelmiin tutustuttiin sekä tehoajolla että vuosihuollon aikana. Nykytilan arvioinnin perusteella esitettiin toimenpide-ehdotuksia irtokappaleiden hallinnan kehittämiseksi. Toimenpide-ehdotukset muodostettiin sekä korjaamaan havaittuja haavoittuvuuksia että kehittämään toimintaa kansainvälisesti hyviksi tunnistettujen käytäntöjen mukaisiksi.

Insulation System in an Integrated Motor Compressor

A high-speed and high-voltage solid-rotor induction machine provides beneficial features for natural gas compressor technology. The mechanical robustness of the machine enables its use in an integrated motor-compressor. The technology uses a centrifugal compressor, which is mounted on the same shaft with the high-speed electrical machine driving it. No gearbox is needed as the speed is determined by the frequency converter. The cooling is provided by the process gas, which flows through the motor and is capable of transferring the heat away from the motor. The technology has been used in the compressors in the natural gas supply chain in the central Europe. New areas of application include natural gas compressors working at the wellheads of the subsea gas reservoir. A key challenge for the design of such a motor is the resistance of the stator insulation to the raw natural gas from the well. The gas contains water and heavy hydrocarbon compounds and it is far harsher than the sales gas in the natural gas supply network. The objective of this doctoral thesis is to discuss the resistance of the insulation to the raw natural gas and the phenomena degrading the insulation. The presence of partial discharges is analyzed in this doctoral dissertation. The breakdown voltage of the gas is measured as a function of pressure and gap distance. The partial discharge activity is measured on small samples representing the windings of the machine. The electrical field behavior is also modeled by finite element methods. Based on the measurements it has been concluded that the discharges are expected to disappear at gas pressures above 4 – 5 bar. The disappearance of discharges is caused by the breakdown strength of the gas, which increases as the pressure increases. Based on the finite element analysis, the physical length of a discharge seen in the PD measurements at atmospheric pressure was approximated to be 40 – 120 m. The chemical aging of the insulation when exposed to raw natural gas is discussed based on a vast set of experimental tests with the gas mixture representing the real gas mixture at the wellhead. The mixture was created by mixing dry hydrocarbon gas, heavy hydrocarbon compounds, monoethylene glycol, and water. The mixture was chosen to be more aggressive by increasing the amount of liquid substances. Furthermore, the temperature and pressure were increased, which resulted in accelerated test conditions. The time required to detect severe degradation was thus decreased. The test program included a comparison of materials, an analysis of the e ects of di erent compounds in the gas mixture, namely water and heavy hydrocarbons, on the aging, an analysis of the e ects of temperature and exposure duration, and also an analysis on the e ect of sudden pressure changes on the degradation of the insulating materials. It was found in the tests that an insulation consisting of mica, glass, and epoxy resin can tolerate the raw natural gas, but it experiences some degradation. The key material in the composite insulation is the resin, which largely defines the performance of the insulation system. The degradation of the insulation is mostly determined by the amount of gas mixture di used into it. The di usion was seen to follow Fick’s second law, but the coe cients were not accurately defined. The di usion was not sensitive to temperature, but it was dependent upon the thermodynamic state of the gas mixture, in other words, the amounts of liquid components in the gas. The weight increase observed was mostly related to heavy hydrocarbon compounds, which act as plasticizers in the epoxy resin. The di usion of these compounds is determined by the crosslink density of the resin. Water causes slight changes in the chemical structure, but these changes do not significantly contribute to the aging phenomena. Sudden changes in pressure can lead to severe damages in the insulation, because the motion of the di used gas is able to create internal cracks in the insulation. Therefore, the di usion only reduces the mechanical strength of the insulation, but the ultimate breakdown can potentially be caused by a sudden drop in the pressure of the process gas.

Incubation period of Hemileia vastatrix in coffee plants in Brazil simulated under climate change

Risk analysis of climate change on plant diseases has great importance for agriculture since it allows the evaluation of management strategies to minimize future damages. This work aimed to simulate future scenarios of coffee rust (Hemileia vastatrix) epidemics by elaborating geographic distribution maps using a model that estimates the pathogen incubation period and the output from three General Circulation Models (CSIRO-Mk3.0, INM-CM3.0, and MIROC3.2.medres). The climatological normal from 1961-1990 was compared with that of the decades 2020s, 2050s and 2080s using scenarios A2 and B1 from the IPCC. Maps were prepared with a spatial resolution of 0.5 × 0.5 degrees of latitude and longitude for ten producing states in Brazil. The climate variables used were maximum and minimum monthly temperatures. The maps obtained in scenario A2 showed a tendency towards a reduction in the incubation period when future scenarios are compared with the climatological normal from 1961-1990. A reduction in the period was also observed in scenario B1, although smaller than that in scenario A2.

Mycobiota evolution during storage of paddy, brown and milled rice in different genotypes

The rice grain is frequently infected by a series of pathogens (fungi) during its storage, producing damages to the economy and health of humans. The aim of this study was to identify the fungal genera present in different rice genotypes and to quantify their variation during storage. Paddy, brown and milled rice fractions of Nutriar, (N) H329-5(H329) and Don Ignacio genotypes were analyzed at 4, 8 and 12 months of storage. Fungi were identified based on their micromorphological characteristics and colonies. The observed genera according to their frequency were: Alternaria, Nigrospora, Epicoccum, Bipolaris, Curvularia, Cladosporium and Fusarium (field fungi) and Penicillium and Aspergillus (storage fungi). The mycobiota composition was different depending on the grain fraction and the period of storage: field fungi were located in the hulls and bran layers, while storage fungi were mainly in the endosperm. The different genotypes showed different susceptibility to contamination.

Sensitivity reduction in Blumeria graminis f. sp. hordei to triadimenol fungicide applied as barley seed treatment

Experiments were carried out in a growth chamber with controlled temperature and photoperiod to test two populations of Blumeria graminis f. sp. hordei from Guarapuava, Paraná State, and Passo Fundo, Rio Grande do Sul State, Brazil. Treatments consisted in application of the fungicide triadimenol (Baytan 150 SC®) at three rates of its commercial formulation: 150, 250, 350 mL/100 Kg barley seeds. The experiments were conducted separately in a growth chamber for each population, adopting the same temperature and photoperiod. For inoculation, pots containing barley seedlings colonized by the fungus were placed among the plots. After emergence of the first symptoms, the disease severity was assessed at two-day intervals. The experiments were repeated twice for each fungus population. Data were expressed as area under the disease progress curve and as powdery mildew control by comparing the severity after the fungicide treatments to that of control. Data were subjected to analysis of variance and regression analysis; the area under the disease progress curve was also calculated. Comparing the data obtained in the present study with those reported in the literature and the control, the maximum value of 26.1% is considered insufficient to prevent the damages caused by the disease. The control response to the fungicide rate was significant. We can conclude that there was a reduction in the sensitivity of both B. graminis f.sp. hordei populations to the fungicide triadimenol, which explains the control failure observed in barley farms.

Etiology of phomopsis root rot in soybean

In a survey of damages caused by soybean root rot to crops in the south of Brazil for several years, a root rot caused by Phomopsis sp has been found with increasing frequency. The primary symptoms are seen when the main root is cut longitudinally, including the death of the wood which shows white coloration and well-defined black lines that do not have a defined format. Thus, based on similarity, it has been called geographic root rot due to its aspect resembling irregular lines that separate regions on a map. In isolations, colonies and alpha spores of Phomopsis have prevailed. Pathogenicity test was done by means of inoculation in the crown of plants cultivated in a growth chamber. The geographic symptoms were reproduced in plants and the fungus Phomopsis sp. was reisolated. In soybean stems naturally infected with pod and stem blight, geographic symptoms caused by Phomopsis phaseoli are found. To the known symptoms on stems, pods and grains, that of root rot caused by P. phaseoli is now added.

DPS-Like Peroxide Resistance Protein: Structural and Functional Studies on a Versatile Nanocontainer

Oxidative stress is a constant threat to almost all organisms. It damages a number of biomolecules and leads to the disruption of many crucial cellular functions. It is caused by reactive oxygen species (ROS), such as hydrogen peroxide (H2O₂), superoxide (•O₂ -), and hydroxyl radical (•OH). The most harmful of these compounds is •OH, which is only formed in cells in the presence of redox-cycling transition metals, such as iron and copper. Bacteria have developed a number of mechanisms to cope with ROS. One of the most widespread means employed by bacteria is the DNA-binding proteins from starved cells (Dps). Dps proteins protect the cells by binding and oxidizing Fe2+, thus greatly reducing the production of •OH. The oxidized iron is stored inside the protein as an iron core. In addition, Dps proteins bind directly to DNA forming a protective coating that shields DNA from harmful agents. Moreover, Dps proteins have been found to elicit other protective functions in cells and to participate in bacterial virulence. Dps proteins are of special importance to Streptococci owing to the lack of catalase in this genus of bacteria.This study was focused on structural and functional characterization of streptococcal Dpslike peroxide resistance (Dpr) proteins. Initially, crystal structures of Streptococcus pyogenes Dpr were determined. The data confirmed the presence of a di-metal ferroxidase center (FOC) in Dpr proteins and revealed the presence of a novel N-terminal helix as well as a surface metal-binding site. The crystal structures of Streptococcus suis Dpr complexed with transition metals demonstrated the metal specificity of the FOC. Solution binding studies also indicated the presence of a di-metal FOC. These results suggested a possible role for Dpr in the detoxification of various metals. Iron was found to mineralize inside the protein as ferrihydrite based on X-ray absorption spectroscopy data. The iron core was found to exhibit clear superparamagnetic behaviour using magnetic and Mössbauer measurements. The results from this study are expected to further increase our understanding on the binding, oxidation, and mineralization of iron and other metals in Dpr proteins. In particular, the structural and magnetic properties of the iron core can form a basis for potential new applications in nanotechnology. From the streptococcal viewpoint, the results would help in understanding better the complicated picture of bacterial pathogenesis. Dpr proteins may also provide a novel target for drug design due to their tight involvement in bacterial virulence.