Sodium/proton exchangers are ubiquitous secondary active transporters that can be found in all kingdoms of life. These proteins facilitate the transport of protons in exchange for sodium ions to help regulate internal pH, sodium levels, and cell volume. Na+/H+ exchangers belong to the SLC9 family and are involved in many physiological processes including cell proliferation, cell migration and vesicle trafficking. Dysfunction of these proteins has been linked to physiological disorders, such as hypertension, heart failure, epilepsy and diabetes. The goal of my thesis is to establish the molecular basis of ion exchange in Na+/H+ exchangers. By establishing how they bind and catalyse the movement of ions across the membrane, we hope we can better understand their role in human physiology. In my thesis, I will first present an overview of Na+/H+ exchangers and their molecular mechanism of ion translocation as was currently understood by structural and functional studies when I started my PhD studies. I will outline our important contributions to this field, which were to (i) obtain the first atomic structures of the same Na+/H+ exchanger (NapA) in two major alternating conformations, (ii) show how a transmembrane embedded lysine residue is essential for carrying out electrogenic transport, and (iii) isolate and recorde the first kinetic data of a mammalian Na+/H+ exchanger (NHA2) in an isolated liposome reconstitution system.

C4 plants have a biochemical carbon concentrating mechanism (CCM) that increases CO2 concentration around Rubisco in the bundle sheath (BS). Maize CCM has two CO2 delivery pathways to the Bundle Sheath (BS) (respectively via malate, MAL or aspartate, ASP); rates of PGA reduction, carbohydrate synthesis and PEP regeneration vary between BS and Mesophyll (M) cells. For these anatomical and biochemical complexities, C4 plants are highly sensitive to light conditions. Under limiting light, the activity of the CCM generally decreases, causing an increase in leakiness, (Φ), the ratio of CO2 retrodiffusing from the BS relative to C4 carboxylation processes. This increase in Φ had been theoretically associated with a decrease in biochemical operating efficiency (expressed as ATP cost of gross assimilation, ATP / GA) under low light and, because a proportion of canopy photosynthesis is carried out by shaded leaves, to potential productivity losses at field scale. In C4 leaves, because of the concentric anatomy, light reaches M cells before the deeper BS (Evans et al., 2007), and could alter the energetic partitioning balance between BS and M and potentially cause efficiency losses. In this experimental programme I investigated strategies deployed by C4 plants to adjust operating efficiency under different illumination conditions. Firstly, maize plants were grown under high and low light regimes (respectively HL, 600 vs LL, 100 μE m-2 s-1). Short term acclimation of Φ was compared from isotopic discrimination (Δ), gas exchange and photochemistry using an improved modelling approach which does not suffer from elements of circularity. Long term acclimation to low light intensities brought about physiological changes which could potentially increase the operating efficiency under limiting ATP supplies. Secondly, profiles of light penetration across a leaf were used to derive the potential ATP supply for M and BS cells induced by changing light quality. Empirical measurements of net CO2 uptake, ATP production rate and carbon isotope discrimination were made on plants under a low light intensity. The overall conversion efficiency was not affected by light quality. A comprehensive metabolic model highlighted the importance of both CO2 delivery pathways in maize. Further, metabolic plasticity allowed the balancing of ATP and NADPH requirements between BS and M. Finally, I tested the hypothesis that plants can modify their physiology so as to reach a status of higher operating efficiency when exposed to high light and then to low light, so as to mimic the transition which leaves undergo when shaded by newly emerging leaves in a crop canopy. Plants were grown under high light and low light for three weeks, then, HL plants were transferred to low light for a further three weeks. Re-acclimation was very effective in reducing ATP cost of net assimilation under low light intensities. In addition, the hyperbolic leakiness increase observed under low light intensities was not associated to operating efficiency loss. Overall, in the three experimental Chapters I showed compelling theoretical and empirical evidence proving the hypothesis that C4 plants deal with low light conditions and with different light qualities without losing operating efficiency.

This research explores the links between the structure and dissociation energetics of ionized non-covalent complexes. In chapter 3, a large series of similar non-covalent complexes were probed using electrospray tandem mass spectrometry (ESI-MS/MS) and RRKM modelling in order to identify any trends in the dissociation energetics based on charge state, overall size of the complex, or size of the substrate. Ion mobility spectrometry (IMS) in conjunction with molecular mechanics/molecular dynamics (MM/MD) was used to study the conformations of these non-covalent complexes in order to determine if the same trends identified in the energetics could be corroborated independently based on structure. The system of study consisted of varying lengths of the synthetic polymer, polymethylmethacrylate (PMMA) complexed with singly or doubly protonated diaminoalkanes (DAA) of varying length. The critical energies of dissociation (E0) increased as the length of the polymer increased and was not significantly affected by the length of the singly protonated DAA substrates. The E0 of dissociation of doubly protonated complexes was strongly influenced by the length of the DAA; longer DAA substrates had greater separation of charge which decreased coulombic repulsion within the complex resulting in higher E0 values. MM/MD low energy structures of all complexes were validated with experimental IMS measurements and showed that the arrangement between the polymer and DAA were similar for different singly protonated DAAs. When doubly protonated, the length of DAA was the most important factor in determining the overall structure of the complex. In chapter 4, a direct link is shown between the observed E0 dissociation energies and the molecular conformations for eight different peptide–saccharide complexes containing either a tri-saccharide (d-(+)-raffinose and d-panose) or tetra-saccharide (stachyose and maltotetraose) with a small peptide (FLEEL and FLEEV). The E0 values were highly related to the overall conformation adopted by the non-covalent complex in the gas phase. Complexes containing peptide FLEE(L/V) with the tri-saccharide raffinose or panose had similar E0 of dissociation (∼0.64 eV) and similar conformations based on MM/MD simulations and IMS drift times. Conversely, for complexes containing a FLEE(L/V) peptide with one of the isomeric tetra-saccharides; stachyose had a E0 ∼0.08 eV greater than maltotetraose. This difference of intermolecular interaction was also reflected by the IMS drift times; maltotetraose in complex with FLEEV or FLEEL had a 5.9% and 2.3% faster IMS drift time than stachyose respectively. This indicated that the molecular arrangement between maltotetraose and the peptides was more compact than the stachyose-peptide complexes. In chapter 5, RRKM modelling of breakdown diagrams is not possible when the reactant ion signal is overlapped by other isobaric species. Trimeric, non-covalent complexes that contained two PMMA molecules and a doubly protonated DAA, [(PMMAa)(DAA+2H)(PMMAb)]+2, have m/z signals that contain multiple different complexes having the same total number of polymer repeat units but differ in the length of the each polymer. In this situation, the applicability of using the simple kinetic method to gain insight into relative binding energies was explored. The major factors which determined the suitability of the kinetic method for this system were identified as the structural arrangement of the reactant ion complex, possible reverse activation barriers, and the evaluations of Δ(ΔS‡). MM/MD simulations coupled with IMS suggests that within the reactant ion, the DAA is almost equally shared between two PMMA oligomers and that the two PMMA oligomers interact predominately with the DAA, and not with each other. MS/MS of the trimeric reactant complexes proceeds by neutral loss of one polymer and is suggested to proceed with little or no reverse activation barrier based on the low coulombic repulsion factors. The IMS drift times of [(PMMAa)(DAA+2H)]+2 complexes that were generated directly by ESI-MS or by dissociation of a trimeric, [(PMMAa)(DAA+2H)(PMMAb)]+2 complex were found to be identical. This provides some evidence that Δ(ΔS‡) ≈ Δ(ΔS) and using a statistical mechanics approach, Δ(ΔS) ≈ 0. The effective temperature (Teff) variable in the kinetic method expression was found to decrease as a function of the size of the trimeric complex, suggesting that the population distribution of the dissociating ensemble of complexes narrows as size increases. Overall, when RRKM fitting is not possible, the simple kinetic method could provide relative energetic ranking of competing dissociations reactions however the Teff term contributed to the greatest uncertainty in obtaining absolute quantities. Fitting MS/MS breakdown diagrams of non-covalent complexes with multiple dissociation channels is difficult due to the number of total fitting variables. Building from the simple kinetic method, chapter 6 shows that the relationship between the natural logarithm of competing fragment ions and reciprocal collision energy yields a branching relationship that allows for the sign of Δ(ΔS‡) and Δ(E0) between the channels to be obtained. Furthermore, the relationships between the fitting variables of RRKM modelling are empirically related to the theoretical branching relationship characteristics. This allowed for the fitting variables of all dissociation channels to be expressed as a function of a single channel so that the theoretical branching relationship matches the experimental branching relationship. Using this method, RRKM fitting of a MS/MS breakdown diagram for APCI ionized anthracene determined the E0 and ∆S‡ was 4.69 ± 0.29 eV and -3 ± 17 J K-1; 4.21 ±0.29 eV and -19 ±15 J K-1; and 4.81 ± 0.29 eV and 36 ±22 J K-1 for hydrogen loss, acetylene loss and diacetylene loss respectively. With one exception, these values are within experimental error of the iPEPICO derived energetic values. In chapter 7, MS/MS of ammoniated triacylglycerides at multiple collision energies and computational analysis are used to explain the cause of uneven dissociation rates of the FAs from different positions on the glycerol backbone. The loss of sn-1 and sn-3 FAs are found to have lower activation energies than the loss of the sn-2 position FA, however the loss of the FA from the sn-2 position is more entropically favourable. Theoretical MS/MS breakdown curves were fit to experimental values using RRKM theory to estimate the E0 of dissociation of FAs from the three glycerol positions. The E0 values for cleavage from the sn-1 and sn-3 positions were found to be approximately 1.52 eV, while that for the sn-2 position was highly dependent on the identity of the FA at that position. Computational structures and energy analysis suggest that an important step in the dissociation of [TAG+NH4]+ is the loss of ammonia. In a model system, glyceryl tributyrate, the loss of NH3 produced two distinct [TAG+H]+ product structures sitting 148 kJ and 160 kJ in energy above the ammoniated structure. The [TAG+H]+ structure that leads to the loss of the sn-1(3) is 12 kJ lower than the [TAG+H]+ structure that leads to the loss of the sn-2 FA. From this, the loss of a neutral FA that follows sits only an additional 35–48 kJ above the [TAG+H]+ structures. In Chapter 8, singly deprotonated β-cyclodextrin monomers, [(β-CD-H+]-1, and doubly deprotonated dimers, [(β-CD)2-2H+]-2, are both present following ESI-MS and have the same monoisotopic m/z. Similar to chapter 5, this makes it difficult to generate an MS/MS breakdown diagrams that can be modelled with RRKM theory. IMS was used to mobility separate [(β-CD-H+]-1 and [(β-CD)2-2H+]-2 and was followed by MS/MS of the [(β-CycD)2-2H+]-2 ion. A second problem when generating a MS/MS breakdown diagram of non-covalent complexes that contain identical components is that the fragment ions could have an identical monoisotopic m/z as the reactant ion. MS/MS of [(β-CycD)2-2H+]-2 results in two [(β-CD-H+]-1 fragments. To overcome this, breakdown diagrams were then generated by monitoring the changes in the isotopic profile. The RRKM derived E0 for dissociation of [(β-CycD)2-H+]-1 and [(β-CycD)2-2H+]-2 were 1.85 ± 0.11eV and 1.79 ± 0.09eV, respectively, corresponding to a slight decrease in complex stability due to increased charge-charge repulsion in the dianion.

The work focuses on wind-power energetics area in the Czech republic. On the basis of a set referential project, which is typical for current Czech conditions, it intends to answer two key questions. The first one deals with risk and inquires into how critical individual factors are. For this purpose financial analysis and especially sensitivity analysis are used. The other issue solved in this work is detection of economic efficiency of the referential project. The output is based on the prediction of the critical factors.

This thesis focuses on innovation of ICT in the energetics sector, particularly in the heating sector. This topic is discussed in terms of both theory and practice. The main goal of this thesis is to show by the example of specific ICT innovations how to properly prepare them. The theoretical part deals with the comparison academic approaches and the approach of the leading Czech supplier of software solutions. It compares the principle of multidimensionality of MMDIS method and methods of Unicorn ES Powered Company. It also discusses in terms of frequency of academic thesis neglected electricity sector, where it focuses on explaining the specifics of this sector using the experience of the author with projects in the Czech and European energetics. It contains a classification of innovation in the industry, market research of ICT products for energetics and SWOT analysis of heat generation sector. The practical part of the thesis focuses on ICT innovation at C -Energy Bohemia. It contains tender documents for this innovation, and based on that is elaborated a methodological approach for evaluation of the benefits of this innovation for the company. Subsequently there is elaborated a proposal for this innovation covering all dimensions of ICT by MMDIS. The main contribution of this work is thus primarily a comparison of academic approaches with the approaches used in practice , the analysis of the current situation of ICT in less well-known electricity industry and the practical application of academic gained from the study of the real demand for ICT innovation.

The thesis deals with the project of the construction of the Temelin Nuclear Power Plant, as an optimal new production source of electrical energy to ensure the majority of the coverage of the growing electric consumption in the Czech Republic and to ensure the state energy independence in the future. The main part of the thesis is dedicated to the methods of strategic investments evaluation. Emphasis is placed on the dynamic methods, which are also working with the factors of time and risk, which are relevant in this case, because the project time horizon is 70 years. The investment project is evaluated from the perspective of owners, as well as from the overall perspective of owners and creditors. The Monte-Carlo method was implemented into the model to support the decision-making process and to move closer to reality. The method assigns the relevant distribution division to the model input values. The output is the set of available values, and the probability of their occurrence. The main thesis objective lies in the decision of the decision maker with a neutral attitude to risk, whether accept the investment or not.

This thesis aims to set a complex view on the economy of the Russian Federation, show its strengths and weaknesses and evaluate its competitiveness in the context of oil and gas reserves. For this purpose the thesis is divided into three chapters. The first one analyses Russian macroeconomics and rates its efficiency, especially after 2000 and in relation to the global financial and economic crisis in 2008. Furthermore, the thesis describes country's energetic sector, its issues and perspectives, impact on economic growth and consequences of insufficient diversification and Russian dependency on export of resources. The main contribution of the thesis represents the last chapter which aims to identify Russian potential with regard to competitiveness, classify its possibilities, suggest solutions and changes needed to enhance competitiveness and ensure sustainable growth independent on resource prices. In conclusion, possible perspectives of Russian development until 2030 are brought forward. The analysis of competitiveness is based on the methodology of the World Economic Forum and Global competitiveness index.

Substantial evidence exists regarding how skeletal muscles use energy and how this affects muscular performance. What remains unclear is how characteristics of muscle energetics affect whole-body energetics during daily living, and what effects this may have on mobility. The goal of this study was to determine the associations between muscle and whole-body energetics including the relationships between: 1) muscle PCr depletion (∆PCr) in response to light intensity isotonic contractions and the oxygen deficit at the onset of a 30-min treadmill walk (30MTW), and, 2) muscle oxidative capacity and excess post-exercise oxygen consumption (EPOC; 30MTW), respiratory exchange ratio (RER; 30MTW), and peak oxygen consumption (VO2 peak) from a graded treadmill test. Eight healthy young (28.4 ± 3.5 years) male participants were studied. Muscle energetics were measured via 31-Phosphorus magnetic resonance spectroscopy (31P-MRS). Muscle ∆PCr was determined as the change in PCr during 2-min of isotonic knee extensor contractions. Muscle oxidative capacity was determined as the rate constant (kPCr) of a PCr recovery following 24-s of maximal isokinetic knee extensor contractions. Whole-body energetic responses to the 30MTW were measured via indirect calorimetry. Oxygen deficit and EPOC were determined as the time constants of the change in oxygen consumption at the onset and offset of the 30MTW, respectively. Respiratory exchange ratio was determined as the mean RER during minutes 7-30 (RER L23), 25-30 (RER L5), and 29-30 (RER L1). Peak oxygen consumption was the highest 30-s average of oxygen consumption during a graded treadmill test, normalized to total mass and lean mass measured by dual-X-ray absorptiometry. Spearman rank correlation coefficients (rs) were calculated to evaluate the associations between independent variables (muscle ∆PCr and oxidative capacity) and dependent variables (oxygen deficit, EPOC, RER, and VO2 peak). Muscle ∆PCr had a positive association (rs = 0.46, p = 0.30) with oxygen deficit. Muscle oxidative capacity had a negative association with EPOC (rs = -0.64, p = 0.14), RER L23 (rs = -0.64, p = 0.14), L5 (rs = -0.68, p = 0.11), and L1 (rs = -0.74, p = 0.07). Muscle oxidative capacity had a positive association with VO2 peak per lean mass (rs = 0.64, p = 0.10), but not VO2 peak per total mass (rs = 0.14, p = 0.75). These results provide promising preliminary evidence that muscle energetics are associated with whole-body energetic response to daily-living type exercise.

This master`s thesis is concerned with a business plan of changing the heating medium. The thesis analyses the current conditions and problems of the company, and offers a business plan, implementation of which will increase the profit of the company and ensure its future growth.

This diploma thesis deals with the history and the development of the gas industry in Czechoslovakia, namely the construction of the transit gas pipeline and development of the gas transit system. One of the part of this thesis is a history of the coal gas and the reasons for its gradual replacement by a natural gas. The metodology of this thesis is based mainly on interviews based on the method of oral history, on archival records, and contemporary literature. Primarily it is focused on oral history fieldwork, particularly on the personal experiences of narrators who were direct participants in the construction. This work is also focused on the circumstances of the negotiations of the construction of a transit gas pipeline and further expansion of the transit system from the seventies to the nineties of the 20th century. Keywords Transit pipeline, natural gas, gas industry, economy, energetics, coal gas, normalization