NADPH-Dependent Thioredoxin System In Regulation of Chloroplast Functions

Photosynthesis, the process in which carbon dioxide is converted into sugars using the energy of sunlight, is vital for heterotrophic life on Earth. In plants, photosynthesis takes place in specific organelles called chloroplasts. During chloroplast biogenesis, light is a prerequisite for the development of functional photosynthetic structures. In addition to photosynthesis, a number of other metabolic processes such as nitrogen assimilation, the biosynthesis of fatty acids, amino acids, vitamins, and hormones are localized to plant chloroplasts. The biosynthetic pathways in chloroplasts are tightly regulated, and especially the reduction/oxidation (redox) signals play important roles in controlling many developmental and metabolic processes in chloroplasts. Thioredoxins are universal regulatory proteins that mediate redox signals in chloroplasts. They are able to modify the structure and function of their target proteins by reduction of disulfide bonds. Oxidized thioredoxins are restored via the action of thioredoxin reductases. Two thioredoxin reductase systems exist in plant chloroplasts, the NADPHdependent thioredoxin reductase C (NTRC) and ferredoxin-thioredoxin reductase (FTR). The ferredoxin-thioredoxin system that is linked to photosynthetic light reactions is involved in light-activation of chloroplast proteins. NADPH can be produced via both the photosynthetic electron transfer reactions in light, and in darkness via the pentose phosphate pathway. These different pathways of NADPH production enable the regulation of diverse metabolic pathways in chloroplasts by the NADPH-dependent thioredoxin system. In this thesis, the role of NADPH-dependent thioredoxin system in the redox-control of chloroplast development and metabolism was studied by characterization of Arabidopsis thaliana T-DNA insertion lines of NTRC gene (ntrc) and by identification of chloroplast proteins regulated by NTRC. The ntrc plants showed the strongest visible phenotypes when grown under short 8-h photoperiod. This indicates that i) chloroplast NADPH-dependent thioredoxin system is non-redundant to ferredoxinthioredoxin system and that ii) NTRC particularly controls the chloroplast processes that are easily imbalanced in daily light/dark rhythms with short day and long night. I identified four processes and the redox-regulated proteins therein that are potentially regulated by NTRC; i) chloroplast development, ii) starch biosynthesis, iii) aromatic amino acid biosynthesis and iv) detoxification of H₂O₂. Such regulation can be achieved directly by modulating the redox state of intramolecular or intermolecular disulfide bridges of enzymes, or by protecting enzymes from oxidation in conjunction with 2-cysteine peroxiredoxins. This thesis work also demonstrated that the enzymatic antioxidant systems in chloroplasts, ascorbate peroxidases, superoxide dismutase and NTRC-dependent 2-cysteine peroxiredoxins are tightly linked up to prevent the detrimental accumulation of reactive oxygen species in plants.

Luminescence-based assay methods in drug discovery

The drug discovery process is facing new challenges in the evaluation process of the lead compounds as the number of new compounds synthesized is increasing. The potentiality of test compounds is most frequently assayed through the binding of the test compound to the target molecule or receptor, or measuring functional secondary effects caused by the test compound in the target model cells, tissues or organism. Modern homogeneous high-throughput-screening (HTS) assays for purified estrogen receptors (ER) utilize various luminescence based detection methods. Fluorescence polarization (FP) is a standard method for ER ligand binding assay. It was used to demonstrate the performance of two-photon excitation of fluorescence (TPFE) vs. the conventional one-photon excitation method. As result, the TPFE method showed improved dynamics and was found to be comparable with the conventional method. It also held potential for efficient miniaturization. Other luminescence based ER assays utilize energy transfer from a long-lifetime luminescent label e.g. lanthanide chelates (Eu, Tb) to a prompt luminescent label, the signal being read in a time-resolved mode. As an alternative to this method, a new single-label (Eu) time-resolved detection method was developed, based on the quenching of the label by a soluble quencher molecule when displaced from the receptor to the solution phase by an unlabeled competing ligand. The new method was paralleled with the standard FP method. It was shown to yield comparable results with the FP method and found to hold a significantly higher signal-tobackground ratio than FP. Cell-based functional assays for determining the extent of cell surface adhesion molecule (CAM) expression combined with microscopy analysis of the target molecules would provide improved information content, compared to an expression level assay alone. In this work, immune response was simulated by exposing endothelial cells to cytokine stimulation and the resulting increase in the level of adhesion molecule expression was analyzed on fixed cells by means of immunocytochemistry utilizing specific long-lifetime luminophore labeled antibodies against chosen adhesion molecules. Results showed that the method was capable of use in amulti-parametric assay for protein expression levels of several CAMs simultaneously, combined with analysis of the cellular localization of the chosen adhesion molecules through time-resolved luminescence microscopy inspection.

Spectrometrical Monitoring of Laser Sintering Process of Ceramic Material

This thesis is done as a part of project called FuncMama that is a project between Technical Research Centre of Finland (VTT), Oulu University (OY), Lappeenranta University of Technology (LUT) and Finnish industrial partners. Main goal of the project is to manufacture electric and mechanical components from mixed materials using laser sintering. Aim of this study was to create laser sintered pieces from ceramic material and monitor the sintering event by using spectrometer. Spectrometer is a device which is capable to record intensity of different wavelengths in relation with time. In this study the monitoring of laser sintering was captured with the equipment which consists of Ocean Optics spectrometer, optical fiber and optical lens (detector head). Light from the sintering process hit first to the lens system which guides the light in to the optical fibre. Optical fibre transmits the light from the sintering process to the spectrometer where wavelengths intensity level information is detected. The optical lens of the spectrometer was rigidly set and did not move along with the laser beam. Data which was collected with spectrometer from the laser sintering process was converted with Excel spreadsheet program for result’s evaluation. Laser equipment used was IPG Photonics pulse fibre laser. Laser parameters were kept mainly constant during experimental part and only sintering speed was changed. That way it was possible to find differences in the monitoring results without fear of too many parameters mixing together and affecting to the conclusions. Parts which were sintered had one layer and size of 5 x 5 mm. Material was CT2000 – tape manufactured by Heraeus which was later on post processed to powder. Monitoring of different sintering speeds was tested by using CT2000 reference powder. Moreover tests how different materials effect to the process monitoring were done by adding foreign powder Du Pont 951 which had suffered in re-grinding and which was more reactive than CT2000. By adding foreign material it simulates situation where two materials are accidently mixed together and it was studied if that can be seen with the spectrometer. It was concluded in this study that with the spectrometer it is possible to detect changes between different laser sintering speeds. When the sintering speed is lowered the intensity level of light is higher from the process. This is a result of higher temperature at the sintering spot and that can be noticed with the spectrometer. That indicates it could be possible to use spectrometer as a tool for process observation and support the idea of having system that can help setting up the process parameter window. Also important conclusion was how well the adding of foreign material could be seen with the spectrometer. When second material was added a significant intensity level raise could be noticed in that part where foreign material was mixed. That indicates it is possible to see if there are any variations in the material or if there are more materials mixed together. Spectrometric monitoring of laser sintering could be useful tool for process window observation and temperature controlling of the sintering process. For example if the process window for specific material is experimentally determined to get wanted properties and satisfying sintering speed. It is possible if the data is constantly recorded that the results can show faults in the part texture between layers. Changes between the monitoring data and the experimentally determined values can then indicate changes in the material being generated by material faults or by wrong process parameters. The results of this study show that spectrometer could be one possible tool for monitoring. But to get in that point where this all can be made possible much more researching is needed.

Microgeneration of electricity with producer gas in dual fuel mode operation

Among the alternatives to meet the increasing of world demand for energy, the use of biomass as energy source is one of the most promising as it contributes to reducing emissions of carbon dioxide in the atmosphere. Gasification is a technological process of biomass energy production of a gaseous biofuel. The fuel gas has a low calorific value that can be used in Diesel engine in dual mode for power generation in isolated communities. This study aimed to evaluate the reduction in the consumption of oil Diesel an engine generator, using gas from gasification of wood. The engine generator brand used was a BRANCO, with direct injection power of 7.36 kW (10 HP) coupled to an electric generator 5.5 kW. Diesel oil mixed with intake air was injected, as the oil was injected via an injector of the engine (dual mode). The fuel gas was produced in a downdraft gasifier. The engine generator was put on load system from 0.5 kW to 3.5 kW through a set of electrical resistances. Diesel oil consumption was measured with a precision scale. It was concluded that the engine converted to dual mode when using the gas for the gasification of wood decreased Diesel consumption by up to 57%.

Thermal comfort temperature update for broiler chickens up to 21 days of age

Due to changes in genetics and nutrition, as well as in acclimatization of broiler chickens to the Brazilian climate, temperature values currently accepted as optimal may be outdated. The objective of this research was to update the environment temperatures that characterize the thermal comfort for broilers chickens from one to 21 days of age, under Brazilian production conditions. This research was conducted with 600 COBB birds, which were distributed in five growth chambers maintained at different temperatures during the first three weeks of age. During the experimental period, temperature values were progressively reduced, consisting in five treatments: T2724/21, T30/27/24, T33/30/27, T36/33/30 and T39/36/33. It was observed that the birds maintained in the T30(27-24) treatment presented better performance compared to other environment conditions. Based on the obtained regression models, the environment temperature values that provide greater weighing gain for the broiler chicken growth in the initial period were 31.3, 25.5 and 21.8 ºC, respectively for the first, second and third week of age.

Start-up of horizontal anaerobic reactors with sludge blanket and fixed bed for wastewater treatment from coffee processing by wet method

In this study it was evaluated the start-up procedures of anaerobic treatment system with three horizontal anaerobic reactors (R1, R2 and R3), installed in series, with volume of 1.2 L each. R1 had sludge blanket, and R2 and R3 had half supporter of bamboo and coconut fiber, respectively. As an affluent, it was synthesized wastewater from mechanical pulping of the coffee fruit by wet method, with a mean value of total chemical oxygen demand (CODtotal) of 16,003 mg L-1. The hydraulic retention time (HRT) in each reactor was 30 h. The volumetric organic loading (VOL) applied in R1 varied from 8.9 to 25.0 g of CODtotal (L d)-1. The mean removal efficiencies of CODtotal varied from 43 to 97% in the treatment system (R1+R2+R3), stabilizing above 80% after 30 days of operation. The mean content of methane in the biogas were of 70 to 76%, the mean volumetric production was 1.7 L CH4 (L reactor d)-1 in the system, and the higher conversions were around at 0.20 L CH4 (g CODremoved)-1 in R1 and R2. The mean values of pH in the effluents ranged from 6.8 to 8.3 and the mean values of total volatile acids remained below 200 mg L-1 in the effluent of R3. The concentrations of total phenols of the affluent ranged from 45 to 278 mg L-1, and the mean removal efficiency was of 52%. The start-up of the anaerobic treatment system occurred after 30 days of operation as a result of inoculation with anaerobic sludge with active microbiota.

Start-up of anaerobic reactors for slaughterhouse wastewater treatment

This study aimed to evaluate the start-up of a horizontal anaerobic fixed bed reactor (HAFBR) followed by an upflow anaerobic sludge blanket (UASB) for the slaughterhouse wastewater treatment. HAFBR was filled with bamboo rings and had 1.2 m in length, 0.10 m in diameter and volume of 7.5 L. The UASB had the volume of 15 L. The HAFBR and UASB operated at organic loading rate and hydraulic retention time average of 8.46 and 3.77 kg m-3 d-1 of COD and 0.53 and 0.98 days, respectively. During 150 days of monitoring system it was found pH 6.8, relatively high values of bicarbonate alkalinity (> 1000 mg L-1) and reduced values of volatile acids (70 to 150 mg L-1), which afforded average removal efficiencies of COD total and total suspended solids of the order of 31 and 23% in HAFBR and 79% and 63% in UASB. It can be concluded that the generation and consumption of bicarbonate alkalinity and total volatile acids, thereby maintaining the pH during the study indicated stable operation of the reactors. The COD removal in the reactors was satisfactory especially when it considers that the assessment was conducted in a period of adaptation of organisms to the effluent and also the high organic load applied during this period.

Finnish Institutional Innovation Support Policies from Russian Entrepreneurs' Perspective: Establishing Cross-Border Technology Oriented Start-Up Firm

The purpose of this thesis is to study how Russian entrepreneurs perceive Finnish institutional environment and innovation support policies provided in the country, as well as to present a practical example in form of a case study of one technology oriented start-up firm which was established in Finland by Russian entrepreneurs. The empirical research of the thesis is conducted qualitatively in two parts. First part is conducted through online questionnaire with open questions in order to review the perceptions of Russian entrepreneurs in general. Second part is based on personal interviews with case company’s founders with the focus on the process of establishing the company in Finland. In the first part of the empirical research, five Russian start-up firms were contacted, and four responses were received. All of these responses were qualified for further analysis. The findings of the first part of the research reveal that Russian entrepreneurs have rather positive attitudes towards Finnish institutional innovation support policies. However, most of the entrepreneurs stated that they are unlikely to create their presence in Finland. As an outcome of the second part of the research, the process of establishing a case company in Finland is illustrated. In order to be able to establish companies in Finland, Russian entrepreneurs who have a permanent residence outside European Economic Area (EEA) are required to apply for a permission to perform business operations in the country. In addition, the established company must engage in improving the economical stand of the country by creating new work places, raising tax revenues, develop technologies and generate innovations in the country.

Promoting peace and security in Africa : is the European Union up to the challenge?

The ninth annual Suomenlinna seminar, organised by the Department of Strategic and Defence Studies, Finnish National Defence College, was held in May 2006. It brought together a representative group of specialists in security studies for two days of presentations and discussion about the most pressing issues of today – the challenges facing European Union’s crisis management in Africa.

Candace Bushnellin Trading Up Edith Whartonin romaanin The Custom of the Country uudelleenkirjoituksena

Candace Bushnellin (1958–) vuonna 2003 julkaistu romaani Trading Up kuvaa nuoren naispäähenkilön määrätietoista pyrkimystä rikkauteen, maineeseen ja New Yorkin suihkuseurapiireihin. Se on uudelleenkirjoitus Edith Whartonin romaanista The Custom of the Country (1913). Wharton (1862–1937) käsitteli tuotannossaan New Yorkin vanhaa yläluokkaa ja kuvasi usein liikettä yhteiskuntaluokassa ylös tai alas. 1990-luvulta alkaen Bushnell on käsitellyt teoksissaan samankaltaisia aiheita, joskin oman aikansa kontekstissa. Hän on itse todennut Whartonin teosten olevan merkittäviä esikuvia omalle tuotannolleen. Pro gradu -tutkielmassani tarkastelen Bushnellin romaania Trading Up Whartonin The Custom of the Country -teoksen uudelleenkirjoituksena. Analysoimalla teoksia rinnakkain selvitän, millaisia keinoja Bushnell käyttää uudelleenkirjoituksessaan ja mikä merkitys Whartonin romaanin uudelleenkirjoituksella on. Tutkimusnäkökulmani on feministinen, ja keskityn valtasuhteisiin liittyviin kysymyksiin. Tarkastelen etenkin yhteiskuntaluokkaan, sukupuoleen ja seksuaalisuuteen liittyviä valtasuhteita ja sitä, miten valtasuhteet Bushnellin uudelleenkirjoituksessa eroavat Whartonin romaanissa esiintyvistä. Tarkastelen tutkimiani romaaneja myös genren näkökulmasta, Trading Up -romaania osana chick lit -genreä ja The Custom of the Country -teosta taparomaanina. Chick lit on 1990- luvulla syntynyt viihteellinen naisten kirjallisuuden genre, joka kuvaa nuoria naisia ja käsittelee esimerkiksi ihmissuhteita ja perheeseen, työelämään tai ulkonäköön liittyviä kysymyksiä. Kulutuskulttuuri ja uusliberalistinen yksilöllisyys kuuluvat chick lit -genreen. Se on kytketty myös postfeminismiin: chick litissä feminismistä joko vaietaan, tai se esitetään pikemminkin yksittäisten naisten elämäntapavalintoina kuin poliittisena liikkeenä. Bushnellin lisäksi myös muut chick lit -kirjailijat ovat teoksissaan viitanneet taparomaaneihin. Tutkielmassani totean Bushnellin uudelleenkirjoittavan The Custom of the Country -romaania paikoitellen hyvin suoraan. Vaikka teosten ajallisen etäisyyden ja yhteiskunnallisten muutosten takia esimerkiksi yhteiskuntaluokkaan liittyvissä valtasuhteissa on tapahtunut muutoksia, esitetään monet asiat muuttumattomina. Etenkin sukupuolen kuvaus ja sukupuoltenväliset valtasuhteet näyttäytyvät samankaltaisina kummassakin teoksessa. Postfeminismissä ja uusliberalismissa yksilön mahdollisuutta tehdä valintoja korostetaan. Bushnellin romaanin naishenkilöhahmoilla pitäisi olla enemmän valinnan mahdollisuuksia kuin Whartonin sankarittarilla, mutta näiden tekemät ratkaisut ovat lopulta hyvin samankaltaisia. Vaikka Trading Up toistaa Whartonin teoksen tarinaa melko tarkasti, ei siitä välity Whartonin romaanissa merkittävää yhteiskuntakriittisyyttä.

Video assisted resections. Increasing access to minimally invasive liver surgery?

Objective: To evaluate perioperative outcomes, safety and feasibility of video-assisted resection for primary and secondary liver lesions. Methods : From a prospective database, we analyzed the perioperative results (up to 90 days) of 25 consecutive patients undergoing video-assisted resections in the period between June 2007 and June 2013. Results : The mean age was 53.4 years (23-73) and 16 (64%) patients were female. Of the total, 84% were suffering from malignant diseases. We performed 33 resections (1 to 4 nodules per patient). The procedures performed were non-anatomical resections (n = 26), segmentectomy (n = 1), 2/3 bisegmentectomy (n = 1), 6/7 bisegmentectomy (n = 1), left hepatectomy (n = 2) and right hepatectomy (n = 2). The procedures contemplated postero-superior segments in 66.7%, requiring multiple or larger resections. The average operating time was 226 minutes (80-420), and anesthesia time, 360 minutes (200-630). The average size of resected nodes was 3.2 cm (0.8 to 10) and the surgical margins were free in all the analyzed specimens. Eight percent of patients needed blood transfusion and no case was converted to open surgery. The length of stay was 6.5 days (3-16). Postoperative complications occurred in 20% of patients, with no perioperative mortality. Conclusion : The video-assisted liver resection is feasible and safe and should be part of the liver surgeon armamentarium for resection of primary and secondary liver lesions.

Predictive Capability of HPV and Pap Tests in Screening for Cervical Cancer over a Three-Year Follow-up

Purpose To compare the predictive capability of HPV and Pap smear tests for screening pre-cancerous lesions of the cervix over a three-year follow-up, in a population of users of the Brazilian National Health System (SUS). Methods This is a retrospective cohort study of 2,032 women with satisfactory results for Pap smear and HPV tests using second-generation hybrid capture,made in a previous study. We followed them for 36 months with data obtained from medical records, the Cervix Cancer Information System (SISCOLO), and the Mortality Information System (SIM). The outcome was a histological diagnosis of cervical intraepithelial neoplasia grade 2 or more advanced lesions (CIN2ş). We constructed progression curves of the baseline test results for the period, using the Kaplan-Meier method, and estimated sensitivity, specificity, positive and negative predictive value, and positive and negative likelihood ratios for each test. Results A total of 1,440 women had at least one test during follow-up. Progression curves of the baseline test results indicated differences in capability to detect CIN2ş (p < 0.001) with significantly greater capability when both tests were abnormal, followed by only a positive HPV test. The HPV test was more sensitive than the Pap smear (88.7% and 73.6%, respectively; p < 0.05) and had a better negative likelihood ratio (0.13 and 0.30, respectively). Specificity and positive likelihood ratio of the tests were similar. Conclusions These findings corroborate the importance of HPV test as a primary cervical cancer screening.