Attenuation of muscle atrophy in a murine model of cachexia by inhibition of the dsRNA-dependent protein kinase

Atrophy of skeletal muscle is due to a depression in protein synthesis and an increase in degradation. Studies in vitro have suggested that activation of the dsRNA-dependent protein kinase (PKR) may be responsible for these changes in protein synthesis and degradation. In order to evaluate whether this is also applicable to cancer cachexia the action of a PKR inhibitor on the development of cachexia has been studied in mice bearing the MAC16 tumour. Treatment of animals with the PKR inhibitor (5 mg kg-1) significantly reduced levels of phospho-PKR in muscle down to that found in non-tumour-bearing mice, and effectively attenuated the depression of body weight, with increased muscle mass, and also inhibited tumour growth. There was an increase in protein synthesis in skeletal muscle, which paralleled a decrease in eukaryotic initiation factor 2α phosphorylation. Protein degradation rates in skeletal muscle were also significantly decreased, as was proteasome activity levels and expression. Myosin levels were increased up to values found in non-tumour-bearing animals. Proteasome expression correlated with a decreased nuclear accumulation of nuclear factor-κB (NF-κB). The PKR inhibitor also significantly inhibited tumour growth, although this appeared to be a separate event from the effect on muscle wasting. These results suggest that inhibition of the autophosphorylation of PKR may represent an appropriate target for the attenuation of muscle atrophy in cancer cachexia. © 2007 Cancer Research UK.

Identification and relative quantification of tyrosine nitration in a model peptide using two-dimensional infrared spectroscopy

Nitration of tyrosine in proteins and peptides is a post-translational modification that occurs under conditions of oxidative stress. It is implicated in a variety of medical conditions, including neurodegenerative and cardiovascular diseases. However, monitoring tyrosine nitration and understanding its role in modifying biological function remains a major challenge. In this work, we investigate the use of electron-vibration-vibration (EVV) two-dimensional infrared (2DIR) spectroscopy for the study of tyrosine nitration in model peptides. We demonstrate the ability of EVV 2DIR spectroscopy to differentiate between the neutral and deprotonated states of 3-nitrotyrosine, and we characterize their spectral signatures using information obtained from quantum chemistry calculations and simulated EVV 2DIR spectra. To test the sensitivity of the technique, we use mixed-peptide samples containing various levels of tyrosine nitration, and we use mass spectrometry to independently verify the level of nitration. We conclude that EVV 2DIR spectroscopy is able to provide detailed spectroscopic information on peptide side-chain modifications and to detect nitration levels down to 1%. We further propose that lower nitration levels could be detected by introducing a resonant Raman probe step to increase the detection sensitivity of EVV 2DIR spectroscopy. (Graph Presented).

Non-Linear Network-Flow Model of Lukasiewicz’s Multivalue Logic

The paper presents a new network-flow interpretation of Łukasiewicz’s logic based on models with an increased effectiveness. The obtained results show that the presented network-flow models principally may work for multivalue logics with more than three states of the variables i.e. with a finite set of states in the interval from 0 to 1. The described models give the opportunity to formulate various logical functions. If the results from a given model that are contained in the obtained values of the arc flow functions are used as input data for other models then it is possible in Łukasiewicz’s logic to interpret successfully other sophisticated logical structures. The obtained models allow a research of Łukasiewicz’s logic with specific effective methods of the network-flow programming. It is possible successfully to use the specific peculiarities and the results pertaining to the function ‘traffic capacity of the network arcs’. Based on the introduced network-flow approach it is possible to interpret other multivalue logics – of E.Post, of L.Brauer, of Kolmogorov, etc.

A proteo-liposome system for the analysis of the intracellular interactome of membrane proteins using amyloid precursor protein as a model

Transmembrane proteins play crucial roles in many important physiological processes. The intracellular domain of membrane proteins is key for their function by interacting with a wide variety of cytosolic proteins. It is therefore important to examine this interaction. A recently developed method to study these interactions, based on the use of liposomes as a model membrane, involves the covalent coupling of the cytoplasmic domains of membrane proteins to the liposome membrane. This allows for the analysis of interaction partners requiring both protein and membrane lipid binding. This thesis further establishes the liposome recruitment system and utilises it to examine the intracellular interactome of the amyloid precursor protein (APP), most well-known for its proteolytic cleavage that results in the production and accumulation of amyloid beta fragments, the main constituent of amyloid plaques in Alzheimer’s disease pathology. Despite this, the physiological function of APP remains largely unclear. Through the use of the proteo-liposome recruitment system two novel interactions of APP’s intracellular domain (AICD) are examined with a view to gaining a greater insight into APP’s physiological function. One of these novel interactions is between AICD and the mTOR complex, a serine/threonine protein kinase that integrates signals from nutrients and growth factors. The kinase domain of mTOR directly binds to AICD and the N-terminal amino acids of AICD are crucial for this interaction. The second novel interaction is between AICD and the endosomal PIKfyve complex, a lipid kinase involved in the production of phosphatidylinositol-3,5-bisphosphate (PI(3,5)P2) from phosphatidylinositol-3-phosphate, which has a role in controlling ensdosome dynamics. The scaffold protein Vac14 of the PIKfyve complex binds directly to AICD and the C-terminus of AICD is important for its interaction with the PIKfyve complex. Using a recently developed intracellular PI(3,5)P2 probe it is shown that APP controls the formation of PI(3,5)P2 positive vesicular structures and that the PIKfyve complex is involved in the trafficking and degradation of APP. Both of these novel APP interactors have important implications of both APP function and Alzheimer’s disease. The proteo-liposome recruitment method is further validated through its use to examine the recruitment and assembly of the AP-2/clathrin coat from purified components to two membrane proteins containing different sorting motifs. Taken together this thesis highlights the proteo-liposome recruitment system as a valuable tool for the study of membrane proteins intracellular interactome. It allows for the mimicking of the protein in its native configuration therefore identifying weaker interactions that are not detected by more conventional methods and also detecting interactions that are mediated by membrane phospholipids.

Em Algorithm for MLE of a Probit Model for Multiple Ordinal Outcomes

The correlated probit model is frequently used for multiple ordered data since it allows to incorporate seamlessly different correlation structures. The estimation of the probit model parameters based on direct maximization of the limited information maximum likelihood is a numerically intensive procedure. We propose an extension of the EM algorithm for obtaining maximum likelihood estimates for a correlated probit model for multiple ordinal outcomes. The algorithm is implemented in the free software environment for statistical computing and graphics R. We present two simulation studies to examine the performance of the developed algorithm. We apply the model to data on 121 women with cervical or endometrial cancer. Patients developed normal tissue reactions as a result of post-operative external beam pelvic radiotherapy. In this work we focused on modeling the effects of a genetic factor on early skin and early urogenital tissue reactions and on assessing the strength of association between the two types of reactions. We established that there was an association between skin reactions and polymorphism XRCC3 codon 241 (C>T) (rs861539) and that skin and urogenital reactions were positively correlated. ACM Computing Classification System (1998): G.3.

Brain-derived neurotrophic factor as an indicator of chemical neurotoxicity:an animal-free CNS cell culture model

Recent changes to the legislation on chemicals and cosmetics testing call for a change in the paradigm regarding the current 'whole animal' approach for identifying chemical hazards, including the assessment of potential neurotoxins. Accordingly, since 2004, we have worked on the development of the integrated co-culture of post-mitotic, human-derived neurons and astrocytes (NT2.N/A), for use as an in vitro functional central nervous system (CNS) model. We have used it successfully to investigate indicators of neurotoxicity. For this purpose, we used NT2.N/A cells to examine the effects of acute exposure to a range of test chemicals on the cellular release of brain-derived neurotrophic factor (BDNF). It was demonstrated that the release of this protective neurotrophin into the culture medium (above that of control levels) occurred consistently in response to sub-cytotoxic levels of known neurotoxic, but not non-neurotoxic, chemicals. These increases in BDNF release were quantifiable, statistically significant, and occurred at concentrations below those at which cell death was measureable, which potentially indicates specific neurotoxicity, as opposed to general cytotoxicity. The fact that the BDNF immunoassay is non-invasive, and that NT2.N/A cells retain their functionality for a period of months, may make this system useful for repeated-dose toxicity testing, which is of particular relevance to cosmetics testing without the use of laboratory animals. In addition, the production of NT2.N/A cells without the use of animal products, such as fetal bovine serum, is being explored, to produce a fully-humanised cellular model.

Interpretation of the Superpave IDT strength test using a viscoelastic-damage constitutive model

This paper presents a new interpretation for the Superpave IDT strength test based on a viscoelastic-damage framework. The framework is based on continuum damage mechanics and the thermodynamics of irreversible processes with an anisotropic damage representation. The new approach introduces considerations for the viscoelastic effects and the damage accumulation that accompanies the fracture process in the interpretation of the Superpave IDT strength test for the identification of the Dissipated Creep Strain Energy (DCSE) limit from the test result. The viscoelastic model is implemented in a Finite Element Method (FEM) program for the simulation of the Superpave IDT strength test. The DCSE values obtained using the new approach is compared with the values obtained using the conventional approach to evaluate the validity of the assumptions made in the conventional interpretation of the test results. The result shows that the conventional approach over-estimates the DCSE value with increasing estimation error at higher deformation rates.

The calculation of dynamic ecological footprint on the basis of the dynamic input-output model

The Leontief input-output model is widely used to determine the ecological footprint of consumption in a region or a country. It is able to capture spillover environmental effects along the supply change, thus its popularity is increasing in ecology related economic research. These studies are static and the dynamic investigations are neglected. The dynamic Leontief model makes it possible to involve the capital and inventory investment in the footprint calculation that projects future growth of GDP and environmental impacts. We show a new calculation method to determine the effect of capital accumulation on ecological footprint. Keywords: Dynamic Leontief model, Dynamic ecological footprint, Environmental management, Allocation method

The Closed Loop Model by Regional Economics or How Big is the Ants’ Footprint?

The economy is communication between Man and Nature. It is an interaction-network between our outside and inside Nature, that is, the external Nature surrounding us and the internal nature expressing our human essence. Money is an institution of the society, an infrastructure that ensures division of labour, enables the flow of information and material between the participants. The concept of regional material and financial circular flow will be more important with the oncoming peak-oil and post-carbon era. We should describe in time the outlines of closed or semi-closed loops economy. The fundamentals of Input-Output will flourish once again; it could help us formulate the link between the efficiency and resiliency of a regional complex system.

A költségvetési korlát megkeményítése a posztszocialista országokban = Hardening of the budget constraint in the post-socialist countries

A költségvetési korlát megkeményítése nem egyforma mértékben ment végbe minden posztszocialista gazdaságban. Egyes országokban messzire jutottak ebben a tekintetben, másokban viszont alig változott az indulóállapot. A tanulmány áttekinti a költségvetési korlát puhaságának különböző megnyilvánulásait: az állami támogatásokat, a puha adózást, a nem teljesítő bankkölcsönöket, a vállalatközi tartozások felgyülemlését és a kifizetetlen béreket. A jelenséget sokféle tényező okozza, amelyek többnyire együttesen jelentkeznek. Az állami tulajdon fenntartása kedvez a puha költségvetési szindróma megrögződésének, a privatizálás elősegíti a keményítést, de nem elégséges feltétele a kemény korlát érvényesítésének. Ehhez megfelelő politikai, jogi és gazdasági környezetet kell céltudatosan kialakítani. A posztszocialista átmenet kezdetén sokan azt hitték, hogy a hatékony piacgazdaság létrehozásához elegendő lesz megvalósítani a liberalizáció, privatizáció és stabilizáció "szentháromságát". Mára már kiderült, hogy a költségvetési korlát megkeményítése az említett három feladattal egyenrangúan fontos. Ahol ez nem valósul meg (például Oroszország), ott a privatizáció nem hozza meg a várt eredményt. ___________________ The budget constraint has not hardened to equal degrees in the various post-socialist countries. In some of them, a great deal has been done in this respect, while in others there has been hardly any change from the initial state. This study surveys the typical manifestations of softness of the budget constraint, such as state subsidies, soft taxation, non-performing loans, the accumulation of trade arrears between firms, and the build-up of wage arrears. Softness of the budget constraint is caused by several factors that tend to act in combination. Thus retention of state ownership helps to preserve the soft budget-constraint syndrome, while privatization encourages the budget constraint to harden, although it does not form a sufficient condition for it to happen. Purposeful development of the requisite political, legal and economic conditions is also required. It was widely maintained at the outset of the post-socialist transition that the 'Holy Trinity' of liberalization, privatization and stabilization would suffice to produce an efficient market economy. Since then, it has become clear that hardening the budget constraint needs to be given equal priority with these. Otherwise, the effects of privatization will fall short of expectations, as they have in Russia, for example.

The effects of using an alternative method for teaching badminton to post secondary level students in Taiwan

The effect of teaching method in physical education is an important issue and has been a concern of the expert teacher. Teachers are expected to create a model of teaching in their field; therefore, it is reasonable to question what is the effect of an alternative teaching method on student performance in physical education. This study explores whether teaching methods with advanced planning, behavior and belief in high enthusiasm, use of instructional strategies and evaluation, together termed a systematic teaching approach, used in a physical education activity would provide an effective environment for learning which supports student achievement in the psychomotor, cognitive, and affective domains. This study also investigated whether there was a difference in performance between students who were taught with a systematic teaching approach and students who were taught with the traditional teaching model. Information was collected using two performance skills, a written test, and one questionnaire. The 68 participants were randomly assigned into either an experimental group or a control group. Two teachers were assigned to either the experimental group or the control group. The teaching experiment took place at Tamsui Oxford University College in Taiwan and lasted eight weeks. ^ Research questions were analyzed using the t-test. Results indicated that a significant difference in students' performance was found between the experimental group and the control group on both the skill tests and the paper test. Analysis of student attitude toward their teacher and their course on the questionnaire indicated a significant difference between the experimental group and the control group. ^ The findings of this study imply that students who were taught with a systematic teaching style were significantly superior to students who were taught with the traditional model on these measures. This finding supports the contention that effective teaching in physical education is related to advanced planning, high enthusiasm, instructional strategy and evaluation and that all physical education teachers should implement these planning elements in the development of the teaching strategies. ^

Late adolescents in transition: The influence of support on post-transition adjustment

Points of transition, when major life roles undergo change, tend to be associated with an increased need for social support. The transition from adolescence to adulthood is ideal for the examination of the effect of normative stress on the development and functioning of social networks. A questionnaire was designed based on the convoy model to assess the influence of personal and situational characteristics on the utilization of support in the prediction of post-transition adjustment. Data were initially collected for a multi-ethnic sample of 741 sophomores and seniors in high school. Surveys were mailed to participants two years later, and one again the following year. The current study is based on data for 310 participants with complete data for all three time periods. A series of hierarchical regressions were conducted to compare three explanatory models of support: main effect, mediation, and moderation. A main effect model of support on post-transition adjustment was confirmed, a mediator model was not confirmed, and a moderator model was marginally confirmed. Family and friend support was related to significantly lower levels of loneliness, particularly for those with less adaptable temperaments. ^

Eulerian-Lagrangian two phase debris flow model

The main objective of this work is to develop a quasi three-dimensional numerical model to simulate stony debris flows, considering a continuum fluid phase, composed by water and fine sediments, and a non-continuum phase including large particles, such as pebbles and boulders. Large particles are treated in a Lagrangian frame of reference using the Discrete Element Method, the fluid phase is based on the Eulerian approach, using the Finite Element Method to solve the depth-averaged Navier-Stokes equations in two horizontal dimensions. The particle’s equations of motion are in three dimensions. The model simulates particle-particle collisions and wall-particle collisions, taking into account that particles are immersed in a fluid. Bingham and Cross rheological models are used for the continuum phase. Both formulations provide very stable results, even in the range of very low shear rates. Bingham formulation is better able to simulate the stopping stage of the fluid when applied shear stresses are low. Results of numerical simulations have been compared with data from laboratory experiments on a flume-fan prototype. Results show that the model is capable of simulating the motion of big particles moving in the fluid flow, handling dense particulate flows and avoiding overlap among particles. An application to simulate debris flow events that occurred in Northern Venezuela in 1999 shows that the model could replicate the main boulder accumulation areas that were surveyed by the USGS. Uniqueness of this research is the integration of mud flow and stony debris movement in a single modeling tool that can be used for planning and management of debris flow prone areas.

An empirical study of Kirkpatrick's evaluation model in the hospitality industry

This study examined Kirkpatrick’s training evaluation model (Kirkpatrick & Kirkpatrick, 2006) by assessing a sales training program conducted at an organization in the hospitality industry. The study assessed the employees’ training outcomes of knowledge and skills, job performance, and the impact of the training upon the organization. By assessing these training outcomes and their relationships, the study demonstrated whether Kirkpatrick’s theories are supported and the lower evaluation levels can be used to predict organizational impact. The population for this study was a group of reservations sales agents from a leading luxury hotel chain’s reservations center. During the study period from January 2005 to May 2007, there were 335 reservations sales agents employed in this Global Reservations Center (GRC). The number of reservations sales agents who had completed a sales training program/intervention during this period and had data available for at least two months pre and post training composed the sample for this study. The number of agents was 69 ( N = 69). Four hypotheses were tested through paired-samples t tests, correlation, and hierarchical regression analytic procedures. Results from the analyses supported the hypotheses in this study. The significant improvement in the call score supported hypothesis one that the reservations sales agents who completed the training improved their knowledge of content and required skills in handling calls (Level 2). Hypothesis two was accepted in part as there was significant improvement in call conversion, but there was no significant improvement of time usage. The significant improvement in the sales per call supported hypothesis three that the reservations agents who completed the training contributed to increased organizational impact (Level 4), i.e., made significantly more sales. Last, findings supported hypothesis four that Level 2 and Level 3 variables can be used for predicting Level 4 organizational impact. The findings supported the theory of Kirkpatrick’s evaluation model that in order to expect organizational results, a positive change in behavior (job performance) and learning must occur. The examinations of Levels 2 and 3 helped to partially explain and predict Level 4 results.

Essays in economic growth, political economy and institutions

This dissertation analyzes the effects of political and economic institutions on economic development and growth.^ The first essay develops an overlapping-generations political economy model to analyze the incentives of various social groups to finance human capital accumulation through public education expenditures. The contribution of this study to the literature is that it helps explain the observed differences in the economic growth performance of natural resource-abundant countries. The results suggest that the preferred tax rates of the manufacturers on one hand and the political coalition of manufacturers and landowners, on the other hand, are equal to the socially optimal tax rate. However, we show that owners of natural resources prefer an excessively high tax rate, which suppresses aggregate output to a suboptimal level.^ The second essay examines the relationship between the political influence of different social classes and public education spending in panel data estimation. The novel contribution of this paper to the literature is that I proxy the political power and influence of the natural resource owners, manufacturers, and landowners with macroeconomic indicators. The motivation behind this modeling choice is to substantiate the definition of the political power of social classes with economic fundamentals. I use different governance indicators in the estimations to find out how different institutions mediate the overall impact of the political influence of various social classes on public education spending. The results suggest that political stability and absence of violence and rule of law are the important governance indicators.^ The third essay develops a counter argument to Acemoglu et al. (2010) where the thesis is that French institutions and economic reforms fostered economic progress in those German regions invaded by the Napoleonic armies. By providing historical data on urbanization rates used as proxies for economic growth, I demonstrate that similar different rates of economic growth were observed in the regions of France in the post-Napoleonic period as well. The existence of different economic growth rates makes it hard to argue that the differences in economic performance in the German regions that were invaded by the French and those that were spared a similar fate follow from regional differences in economic institutions.^