Reversal of a full-length mutant huntingtin neuronal cell phenotypeby chemical inhibitors of polyglutamine-mediated aggregation

Background: Huntington's disease (HD) is an inherited neurodegenerative disorder triggered by an expanded polyglutamine tract in huntingtin that is thought to confer a new conformational property on this large protein. The propensity of small amino-terminal fragments with mutant, but not wild-type, glutamine tracts to self-aggregate is consistent with an altered conformation but such fragments occur relatively late in the disease process in human patients and mouse models expressing full-length mutant protein. This suggests that the altered conformational property may act within the full-length mutant huntingtin to initially trigger pathogenesis. Indeed, genotypephenotype studies in HD have defined genetic criteria for the disease initiating mechanism, and these are all fulfilled by phenotypes associated with expression of full-length mutant huntingtin, but not amino-terminal fragment, in mouse models. As the in vitro aggregation of amino-terminal mutant huntingtin fragment offers a ready assay to identify small compounds that interfere with the conformation of the polyglutamine tract, we have identified a number of aggregation inhibitors, and tested whether these are also capable of reversing a phenotype caused by endogenous expressionof mutant huntingtin in a striatal cell line from the HdhQ111/Q111 knock-in mouse. Results: We screened the NINDS Custom Collection of 1,040 FDA approved drugs and bioactive compounds for their ability to prevent in vitro aggregation of Q58-htn 1¿171 amino terminal fragment. Ten compounds were identified that inhibited aggregation with IC50 < 15 ¿M, including gossypol, gambogic acid, juglone, celastrol, sanguinarine and anthralin. Of these, both juglone and celastrol were effective in reversing the abnormal cellular localization of full-length mutant huntingtin observed in mutant HdhQ111/Q111 striatal cells. Conclusions: At least some compounds identified as aggregation inhibitors also prevent a neuronal cellular phenotype caused by full-length mutant huntingtin, suggesting that in vitro fragment aggregation can act as a proxy for monitoring the disease-producing conformational property in HD. Thus, identification and testing of compounds that alter in vitro aggregation is a viable approach for defining potential therapeutic compounds that may act on the deleterious conformational property of full-length mutant huntingtin.

Discrepancies over the onset of surfactant monomer aggregation interpreted by fluorescence, conductivity and surface tension methods

Molecular probe techniques have made important contributions to the determination of microstructure of surfactant assemblies such as size, stability, micropolarity and conformation. Conductivity and surface tension were used to determine the critical aggregation concentration (cac) of polymer-surfactant complexes and the critical micellar concentration (cmc) of aqueous micellar aggregates. The results are compared with those of fluorescent techniques. Several surfactant systems were examined, 1-butanol-sodium dodecylsulfate (SDS) mixtures, solutions containing poly(ethylene oxide)-SDS, poly(vinylpyrrolidone)-SDS and poly(acrylic acid)-alkyltrimethylammonium bromide complexes. We found differences between the cac and cmc values obtained by conductivity or surface tension and those obtained by techniques which use hydrophobic probe.

Many valued algebraic structures as measures of comparison

This thesis studies the properties and usability of operators called t-norms, t-conorms, uninorms, as well as many valued implications and equivalences. Into these operators, weights and a generalized mean are embedded for aggregation, and they are used for comparison tasks and for this reason they are referred to as comparison measures. The thesis illustrates how these operators can be weighted with a differential evolution and aggregated with a generalized mean, and the kinds of measures of comparison that can be achieved from this procedure. New operators suitable for comparison measures are suggested. These operators are combination measures based on the use of t-norms and t-conorms, the generalized 3_-uninorm and pseudo equivalence measures based on S-type implications. The empirical part of this thesis demonstrates how these new comparison measures work in the field of classification, for example, in the classification of medical data. The second application area is from the field of sports medicine and it represents an expert system for defining an athlete's aerobic and anaerobic thresholds. The core of this thesis offers definitions for comparison measures and illustrates that there is no actual difference in the results achieved in comparison tasks, by the use of comparison measures based on distance, versus comparison measures based on many valued logical structures. The approach has been highly practical in this thesis and all usage of the measures has been validated mainly by practical testing. In general, many different types of operators suitable for comparison tasks have been presented in fuzzy logic literature and there has been little or no experimental work with these operators.

Heterogeneous products and tests for an appropriate aggregation level considering different qualities: evidence from fresh hake at Barcelona’s wholesale market

[cat] Aquest estudi destaca la importància de considerar un nivell d’agregació adequat en els anàlisis de demanda, ja que treballar utilitzant un nivell d’agregació inadequat pot donar lloc a estimacions esbiaixades. Aquest fet es mostra a través de l’anàlisi de diferents productes de lluç fresc comercialitzats a Mercabarna, el mercat majorista de Barcelona. La literatura sobre la demanda de peix tracta al lluç com un únic producte i espècie. No obstant això, en el mercat espanyol, es comercialitzen molts peixos com a lluç, els quals mostren comportaments molt diferents (des de béns inferiors fins a béns de luxe). Els resultats obtinguts, en concordança amb les observacions empíriques, demostren que l’anàlisi s’ha de realitzar amb un major grau de detall que a nivell d’espècie. Això qüestiona els resultats d’anteriors estudis de demanda i la majoria de les bases de dades, on l’observació del nivell d’agregació adequat dels productes no es té en compte.

Re-visiting the health care luxury good hypothesis: Aggregation, precision, and publication biases?

[cat] Mentre que una creixent literatura que ha examinat la relació entre la renda i la despesa sanitària suggereix que els serveis sanitaris són un be de luxe (elasticitat renda superior a la unitat), aquesta conclusió es contínuament debatuda atesa l'heterogeneïtat dels resultats. Aquest article testa la hipòtesis dels serveis sanitaris com bens de luxe fent server anàlisi de meta- regressió, particularment analitzant l'existència de biaixos de selecció de publicació, precisió així com biaixos d'agregació. Els resultats apunten l'existència d'un biaix de publicació, robust independentment dels controls analitzats. Els biaixos de precisió i agregació semblen tenir un paper en la generació de les estimacions de l'elasticitat renda. Els nostres resultat suggereixen que l'elasticitat renda dels serveis sanitaris un cop corregir pels biaixos esmentat varien entre 0.26 i 0.84, però no podem rebutjar que la elasticitat renda es igual a la unitat en algunes estimacions de l'elasticitat corregides.

Heterogeneous products and tests for an appropriate aggregation level considering different qualities: evidence from fresh hake at Barcelona’s wholesale market

[cat] Aquest estudi destaca la importància de considerar un nivell d’agregació adequat en els anàlisis de demanda, ja que treballar utilitzant un nivell d’agregació inadequat pot donar lloc a estimacions esbiaixades. Aquest fet es mostra a través de l’anàlisi de diferents productes de lluç fresc comercialitzats a Mercabarna, el mercat majorista de Barcelona. La literatura sobre la demanda de peix tracta al lluç com un únic producte i espècie. No obstant això, en el mercat espanyol, es comercialitzen molts peixos com a lluç, els quals mostren comportaments molt diferents (des de béns inferiors fins a béns de luxe). Els resultats obtinguts, en concordança amb les observacions empíriques, demostren que l’anàlisi s’ha de realitzar amb un major grau de detall que a nivell d’espècie. Això qüestiona els resultats d’anteriors estudis de demanda i la majoria de les bases de dades, on l’observació del nivell d’agregació adequat dels productes no es té en compte.

Re-visiting the health care luxury good hypothesis: Aggregation, precision, and publication biases?

[cat] Mentre que una creixent literatura que ha examinat la relació entre la renda i la despesa sanitària suggereix que els serveis sanitaris són un be de luxe (elasticitat renda superior a la unitat), aquesta conclusió es contínuament debatuda atesa l'heterogeneïtat dels resultats. Aquest article testa la hipòtesis dels serveis sanitaris com bens de luxe fent server anàlisi de meta- regressió, particularment analitzant l'existència de biaixos de selecció de publicació, precisió així com biaixos d'agregació. Els resultats apunten l'existència d'un biaix de publicació, robust independentment dels controls analitzats. Els biaixos de precisió i agregació semblen tenir un paper en la generació de les estimacions de l'elasticitat renda. Els nostres resultat suggereixen que l'elasticitat renda dels serveis sanitaris un cop corregir pels biaixos esmentat varien entre 0.26 i 0.84, però no podem rebutjar que la elasticitat renda es igual a la unitat en algunes estimacions de l'elasticitat corregides.

The governmental organization's role in railway freight market's liberalization -building knowledge through Swedish and Polish operators' experiences

Globalization has increased transport aggregates’ demand. Whilst transport volumes increase, ecological values’im portance has sharpened: carbon footprint has become a measure known world widely. European Union together with other communities emphasizes friendliness to the environment: same trend has extended to transports. As a potential substitute for road transport is noted railway transport, which decreases the congestions and lowers the emission levels. Railway freight market was liberalized in the European Union 2007, which enabled new operators to enter the markets. This research had two main objectives. Firstly, it examined the main market entry strategies utilized and the barriers to entry confronted by the operators who entered the markets after the liberalization. Secondly, the aim was to find ways the governmental organization could enhance its service towards potential railway freight operators. Research is a qualitative case study, utilizing descriptive analytical research method with a normative shade. Empirical data was gathered by interviewing Swedish and Polish railway freight operators by using a semi-structured theme-interview. This research provided novel information by using first-hand data; topic has been researched previously by utilizing second-hand data and literature analyses. Based on this research, rolling stock acquisition, needed investments and bureaucracy generate the main barriers to entry. The research results show that the mostly utilized market entry strategies are start-up and vertical integration. The governmental organization could enhance the market entry process by organizing courses, paying extra attention on flexibility, internal know-how and educating the staff.

Thioflavin-S staining of bacterial inclusion bodies for the fast, simple, and inexpensive screening of amyloid aggregation inhibitors

Amyloid aggregation is linked to a large number of human disorders, from neurodegenerative diseases as Alzheimer"s disease (AD) or spongiform encephalopathies to non-neuropathic localized diseases as type II diabetes and cataracts. Because the formation of insoluble inclusion bodies (IBs) during recombinant protein production in bacteria has been recently shown to share mechanistic features with amyloid self-assembly, bacteria have emerged as a tool to study amyloid aggregation. Herein we present a fast, simple, inexpensive and quantitative method for the screening of potential anti-aggregating drugs. This method is based on monitoring the changes in the binding of thioflavin-S to intracellular IBs in intact Eschericchia coli cells in the presence of small chemical compounds. This in vivo technique fairly recapitulates previous in vitro data. Here we mainly use the Alzheimer"s related beta-amyloid peptide as a model system, but the technique can be easily implemented for screening inhibitors relevant for other conformational diseases simply by changing the recombinant amyloid protein target. Indeed, we show that this methodology can be also applied to the evaluation of inhibitors of the aggregation of tau protein, another amyloidogenic protein with a key role in AD.

Dual inhibitors of beta-amyloid aggregation and acetylcholinesterase as multi-target anti-Alzheimer drug candidates

Notwithstanding the functional role that the aggregates of some amyloidogenic proteins can play in different organisms, protein aggregation plays a pivotal role in the pathogenesis of a large number of human diseases. One of such diseases is Alzheimer"s disease (AD), where the overproduction and aggregation of the β-amyloid peptide (Aβ) are regarded as early critical factors. Another protein that seems to occupy a prominent position within the complex pathological network of AD is the enzyme acetylcholinesterase (AChE), with classical and non-classical activities involved at the late (cholinergic deficit) and early (Aβ aggregation) phases of the disease. Dual inhibitors of Aβ aggregation and AChE are thus emerging as promising multi-target agents with potential to efficiently modify the natural course of AD. In the initial phases of the drug discovery process of such compounds, in vitro evaluation of the inhibition of Aβ aggregation is rather troublesome, as it is very sensitive to experimental assay conditions, and requires expensive synthetic Aβ peptides, which makes cost-prohibitive the screening of large compound libraries. Herein, we review recently developed multi-target anti-Alzheimer compounds that exhibit both Aβ aggregation and AChE inhibitory activities, and, in some cases also additional valuable activities such as BACE-1 inhibition or antioxidant properties. We also discuss the development of simplified in vivo methods for the rapid, simple, reliable, unexpensive, and high-throughput amenable screening of Aβ aggregation inhibitors that rely on the overexpression of Aβ42 alone or fused with reporter proteins in Escherichia coli.

Thioflavin-S staining of bacterial inclusion bodies for the fast, simple, and inexpensive screening of amyloid aggregation inhibitors

Amyloid aggregation is linked to a large number of human disorders, from neurodegenerative diseases as Alzheimer"s disease (AD) or spongiform encephalopathies to non-neuropathic localized diseases as type II diabetes and cataracts. Because the formation of insoluble inclusion bodies (IBs) during recombinant protein production in bacteria has been recently shown to share mechanistic features with amyloid self-assembly, bacteria have emerged as a tool to study amyloid aggregation. Herein we present a fast, simple, inexpensive and quantitative method for the screening of potential anti-aggregating drugs. This method is based on monitoring the changes in the binding of thioflavin-S to intracellular IBs in intact Eschericchia coli cells in the presence of small chemical compounds. This in vivo technique fairly recapitulates previous in vitro data. Here we mainly use the Alzheimer"s related beta-amyloid peptide as a model system, but the technique can be easily implemented for screening inhibitors relevant for other conformational diseases simply by changing the recombinant amyloid protein target. Indeed, we show that this methodology can be also applied to the evaluation of inhibitors of the aggregation of tau protein, another amyloidogenic protein with a key role in AD.

Conservation laws for non-potential operators

The aim of this work is to study the conservation laws of continuous means mechanics and also to extend the Hamiltonian method for these kind of systems in order to valid for non-potential operators through variational approach. Besides illustrating with various examples of mechanical applications we also introduce in this work the new technique in order to treat such problems as the non-potential problem.

Railways, Airports and Sea Container Operators as Publicly Listed Companies – Financial Performance and Shareholder Value Creation Perspective

Logistics infrastructure and transportation services have been the liability of countries and governments for decades, or these have been under strict regulation policies. One of the first branches opened for competition in EU as well as in other continents, has been air transports (operators, like passenger and freight) and road transports. These have resulted on lower costs, better connectivity and in most of the cases higher service quality. However, quite large amount of other logistics related activities are still directly (or indirectly) under governmental influence, e.g. railway infrastructure, road infrastructure, railway operations, airports, and sea ports. Due to the globalization, governmental influence is not that necessary in this sector, since transportation needs have increased with much more significant phase as compared to economic growth. Also freight transportation needs do not correlate with passenger side, due to the reason that only small number of areas in the world have specialized in the production of particular goods. Therefore, in number of cases public-private partnership, or even privately owned companies operating in these sub-branches have been identified as beneficial for countries, customers and further economic growth. The objective of this research work is to shed more light on these kinds of experiments, especially in the relatively unknown sub-branches of logistics like railways, airports and sea container transports. In this research work we have selected companies having public listed status in some stock exchange, and have needed amount of financial scale to be considered as serious company rather than start-up phase venture. Our research results show that railways and airports usually need high fixed investments, but have showed in the last five years generally good financial performance, both in terms of profitability and cash flow. In contrary to common belief of prosperity in globally growing container transports, sea vessel operators of containers have not shown that impressive financial performance. Generally margins in this business are thin, and profitability has been sacrificed in front of high growth – this also concerns cash flow performance, which has been lower too. However, as we examine these three logistics sub-branches through shareholder value development angle during time period of 2002-2007, we were surprised to find out that all of these three have outperformed general stock market indexes in this period. More surprising is the result that financially a bit less performing sea container transportation sector shows highest shareholder value gain in the examination period. Thus, it should be remembered that provided analysis shows only limited picture, since e.g. dividends were not taken into consideration in this research work. Therefore, e.g. US railway operators have disadvantage to other in the analysis, since they have been able to provide dividends for shareholders in long period of time. Based on this research work we argue that investment on transportation/logistics sector seems to be safe alternative, which yields with relatively low risk high gain. Although global economy would face smaller growth period, this sector seems to provide opportunities in more demanding situation as well.