Conditioned media prepared from umbilical cord (UC) segments or endothelial cells (EC) contain colony stimulating activity, Both UCCM and ECCM were partially purified by DEAE-Sepharose and ACA44 gel filtration chromatography. The molecular weights were estimated as 25,000 and 31,000 for UC-CSF and EC-CSF, respectively. UC-CSF was further fractionated by Con A Sepharose, IEF and HPLC on a hydrophobic phenyl column. The highly purified CSF stimulates human macrophage and granulocyte colony formation, indicating it is GM-CSF in nature. Characterization studies have revealed that both CSFs are heat stable at 60°C for 30 min. They are sensitive to digestion by protease and to periodate oxidation but are stable to treatment with sulfhydryl reagents. The synthesis of CSF in endothelial cells is inhibited by actinomycin D, cycloheximide and puromycin, indicating that protein and RNA synthesis are required for CSF production. Among the mitogens tested, only LPS exhibited stimulatory activity on the production of CSF. Metabolic modulators such as dibutyryl cAMP, isobutylmethylxanthine, PGE2 and lactoferrin inhibit CSF production, while PGF2 enhances CSF production.

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Combinatorial optimization problems are of high academical as well as practical importance. Many instances of relevant combinatorial optimization problems are, due to their dimensions, intractable for complete methods such as branch and bound. Therefore, approximate algorithms such as metaheuristics received much attention in the past 20 years. Examples of metaheuristics are simulated annealing, tabu search, and evolutionary computation. One of the most recent metaheuristics is ant colony optimization (ACO), which was developed by Prof. M. Dorigo (who is the supervisor of this thesis) and colleagues. This thesis deals with theoretical as well as practical aspects of ant colony optimization. * A survey of metaheuristics. Chapter 1 gives an extensive overview on the nowadays most important metaheuristics. This overview points out the importance of two important concepts in metaheuristics: intensification and diversification. * The hyper-cube framework. Chapter 2 introduces a new framework for implementing ACO algorithms. This framework brings two main benefits to ACO researchers. First, from the point of view of the theoretician: we prove that Ant System (the first ACO algorithm to be proposed in the literature) in the hyper-cube framework generates solutions whose expected quality monotonically increases with the number of algorithm iterations when applied to unconstrained problems. Second, from the point of view of the experimental researcher, we show through examples that the implementation of ACO algorithms in the hyper-cube framework increases their robustness and makes the handling of the pheromone values easier. * Deception. In the first part of Chapter 3 we formally define the notions of first and second order deception in ant colony optimization. Hereby, first order deception corresponds to deception as defined in the field of evolutionary computation and is therefore a bias introduced by the problem (instance) to be solved. Second order deception is an ACO-specific phenomenon. It describes the observation that the quality of the solutions generated by ACO algorithms may decrease over time in certain settings. In the second part of Chapter 3 we propose different ways of avoiding second order deception. * ACO for the KCT problem. In Chapter 4 we outline an ACO algorithm for the edge-weighted k-cardinality tree (KCT) problem. This algorithm is implemented in the hyper-cube framework and uses a pheromone model that was determined to be well-working in Chapter 3. Together with the evolutionary computation and the tabu search approaches that we develop in Chapter 4, this ACO algorithm belongs to the current state-of-the-art algorithms for the KCT problem. * ACO for the GSS problem. Chapter 5 describes a new ACO algorithm for the group shop scheduling (GSS) problem, which is a general shop scheduling problem that includes among others the well-known job shop scheduling (JSS) and the open shop scheduling (OSS) problems. This ACO algorithm, which is implemented in the hyper-cube framework and which uses a new pheromone model that was experimentally tested in Chapter 3, is currently the best ACO algorithm for the JSS as well as the OSS problem. In particular when applied to OSS problem instances, this algorithm obtains excellent results, improving the best known solution for several OSS benchmark instances. A final contribution of this thesis is the development of a general method for the solution of combinatorial optimization problems which we refer to as Beam-ACO. This method is a hybrid between ACO and a tree search technique known as beam search. We show that Beam-ACO is currently a state-of-the-art method for the application to the existing open shop scheduling (OSS) problem instances.

In a multiuser system, several signals are transmitted simultaneously within the same frequency band. This can result in significant improvements both in spectral efficiency and system capacity. However, a detrimental effect of the shared transmissions (both in time and bandwidth), is that the signal received at the base station (BS) or access point (AP) suffers from cochannel interference (CCI) and inter-symbol interference (ISI). This situation presents challenges to receiver design. To combat the destructive nature of multipath fading, a receiver often employs multiple antennas to collect the faded superimposed versions of the transmitted signals. The multiple signals are combined and processed in such a way that the effects of CCI and ISI are minimized and the desired information is reliably recovered. The situation is even more challenging when the system is operating under overload, i.e. when there are fewer receive antennas than there are transmitted signals. Multiuser detection (MUD) is used to simultaneously estimate the information sent by the transmitters. To do this, the receiver exploits differences among the cochannel signals (through unique spatial signatures in this case). We consider a cochannel communication system where multiple transmitted signals arrive at a receiver (equipped with multiple receive antennas) after propagating through a Rayleigh fading channel. It is assumed that the receiver is operating in an overloaded scenario. For such systems, an optimum maximum a posterior probability (MAP) detector estimates the transmitted signal by maximizing the probability of correct decision. The MAP detector reduces to the maximum likelihood (ML) detector when all the transmitted signals are equiprobable. The computational complexity of both MAP and ML detectors increases exponentially with the number of transmitted signals and the channel memory. For large systems suffering severe CCI and ISI, this is clearly not a good choice for real-time implementation due to the associated computational expenses. The main factors that influence the complexity of MAP / ML detection are: (i) the number of transmitted signals (or equivalently the number of users sharing the system resources), (ii) modulation alphabet size, and (iii) length of the channel memory. On the other hand, linear detection approaches fail to offer acceptable performance while other nonlinear sub-optimum approaches incur high computational costs for reasonably improved system performance and exhibit an irreducible error-floor at medium to high signal to noise ratio (SNR) values. We develop receiver signal processing techniques for the frequency-flat fading channel (where all the multipaths of the transmitted signal arrive at the receiver within a symbol period). We develop an ant colony optimization (ACO) assisted soft iterative detection approach for binary phase-shift keying (BPSK) modulated signals which employs a simplified MAP criteria to extract the most probable signals from the search space. The structure of the receiver is such that it can continue operating under overloaded conditions. The technique achieves near maximum likelihood (ML) performance in critically loaded cases using much lower complexity. For the challenging case of overload it still offers performance close to ML at low to moderate SNR values. Second, an integrated framework comprising of ACO metaheuristic and a recursively defined ML search criteria is developed to handle multilevel modulations. The proposed receiver is capable of achieving near-ML performance for the considered system with significant savings in computational complexity. The receiver framework is independent of the system loading condition, and therefore it remains suitable for overloaded scenarios. Due to the branch and bound nature of the algorithm, an exact expression for the complexity cannot be determined. Instead, an upper bound on computational complexity is developed.

The colony formation assay (CFA) is the gold standard for measuring cytotoxic effects on cells. To increase efficiency, the CFA was converted to a 96-well format using an automated colony counting algorithm. The 96-well CFA was validated using ionizing radiation (IR) on the FaDu and A549 cancer cell lines. Its ability to evaluate combination therapies was investigated using cisplatin and IR. The 96-well CFA was transferred to a robotic platform for evaluation as a high-throughput screen (HTS) readout for the discovery of novel anti-cancer compounds, and radiosensitizers. Screening yielded eight putative anti-cancer hits, and five putative radiosensitizing hits. Secondary screening confirmed 6/8 anti-cancer compounds, and 0/5 radiosensitizing compounds. Thus, the 96-well CFA can be adopted as an alternative assay to the 6-well CFA in the evaluation of cytotoxicity in vitro, providing a possible readout to be utilized in HTS for discovering anti-cancer compounds, but with limited applicability in discovering radiosensitizers.

Calcium is a highly versatile intracellular signal molecule that can regulate numerous different cellular functions. In filamentous fungi there is evidence for it being involved in regulating various processes, including spore germination, hyphal tip growth, hyphal branching and conidiation. During colony initiation in the filamentous fungus Neurospora crassa, conidia form germ tubes which are involved in colony establishment, and conidial anastomosis tubes (CATs) which are involved in generating fused networks of conidial germlings. The primary research aim of this thesis was to analyze the role of Ca2+-signalling and homeostasis during colony initiation in N. crassa. Removal of Ca2+ from the growth medium showed that external Ca2+ was necessary for CAT fusion and, more specifically, was required for CAT chemoattraction. Two L-type Ca2+ channel blockers (verapamil and diltiazem) with different modes of action were found to inhibit both conidial germination and CAT fusion in wild type strains and CAT fusion was shown to be more sensitive to these two drugs. These channel blockers were additionally found to inhibit Ca2+ uptake by conidial germlings of the wild type expressing the aequorin Ca2+ reporter. However, the channel blockers also, unexpectedly, raised the cytosolic free Ca2+ ([Ca2+]c) resting level in these germlings suggesting that they did not just inhibit L-type Ca2+ activity. The morphological phenotypes (conidial germination, hyphal extension rate, conidiation and hyphal branching) of 22 mutants defective in different components of their Ca2+-signalling and homeostasis machinery were characterized in order to identify their possible roles of Ca2+ during colony initiation and development. The ∆cch-1 mutant lacking the CCH-1 L-type Ca2+ channel gene exhibited a reduction in CAT fusion. CAT fusion was decreased even further in a double mutant (∆cch-1∆mid-1) suggesting that that the CCH-1 and MID-1 proteins operate in combination during this process. Increased extracellular Ca2+ partially restored the phenotypes of the ∆cch-1, ∆mid-1 smco-1 and ∆cch-1∆mid-1 mutants which is consistent with CCH-1 and MID-1 being involved in Ca2+ uptake from the external medium. Calcium signatures following mechanical perturbation were successfully measured in populations of conidial germlings using aequorin expressed in the wild type and in deletion mutants (∆cch-1, ∆yvc-1, ∆fig-1) lacking different Ca2+ channels. The removal of external Ca2+ completely abolished the [Ca2+]c increase in response to mechanical perturbation and CCH-1 was found to partly contribute to this increase in [Ca2+]c. Various Ca2+-sensitive dyes (Oregon green 488, Fluo-4 and Calcium Green-1) were also tested to determine if they can be used to image [Ca2+]c at the single cell and subcellular levels. Only Fluo-4 allowed the measurement of [Ca2+]c in individual cells but the changes in dye fluorescence in response to changes in [Ca2+]c were too small to be useful for imaging [Ca2+]c dynamics at the subcellular level. The other two dyes underwent rapid compartmentalization in organelles when loaded into germlings. The plant antifungal proteins (defensins), MsDef1, MtDef4 and PAF were all found to disrupt Ca2+ signaling/homeostasis in conidial germlings of N. crassa. They all inhibited the [Ca2+]c increase and raised the resting level of [Ca2+]c in response to mechanical perturbation. Analysis of an aequorin expressing mutant that was defective in glucosylceramide synthase (∆gcs) showed that the effects of MsDef1 (but not MtDef4) on [Ca2+]c were mediated by the sphingolipid glucosylceramide. All of the defensins tested were found to exhibit different potencies with regard to their inhibitory effects on conidial germination and CAT fusion.

De 1896 à 1960, quoique placée sous différents statuts (colonie, territoire d'outre-mer, puis État membre de la Communauté), Madagascar est restée sous la domination française. Durant cette période, la France administre le pays, notamment, elle doit faire régner la justice, un des premiers éléments de la mission du colonisateur. A cet égard, la politique consiste à introduire dans l'organisation judiciaire des magistrats français qui, en charge de véhiculer les valeurs occidentales, vont assurément participer de la domination. Toutefois, en même temps, en exerçant sur le territoire malgache, ces juges se heurtent à l'existence d'institutions indigènes déjà bien établies qu'il convient de respecter. Par ailleurs, ils se trouvent face à une population européenne nouvellement installée à qui il faut aussi « dire » le droit, un droit somme toute particulier. Ainsi, au sein de la Cour d'appel de Madagascar, les magistrats jouent un rôle majeur en ce qu'ils ont à remplir une triple mission à la fois : appliquer un droit spécial aux Français, appliquer leurs lois coutumières aux indigènes et appliquer le droit métropolitain, « symbole éminent de la culture française », à la population locale. Dans la pratique, malgré les difficultés qu'ils ont rencontrées outre-mer, et les contraintes qui se sont imposées à eux, les magistrats s'en sont plutôt bien sortis dans la délicate tâche qui leur a été confiée. En se dotant, sans complexe, de toute la liberté nécessaire à leur fonction, ils ont su, à leur façon, satisfaire « leurs » justiciables tout en restant fidèle à la politique coloniale. / From 1896 to 1960, Madagascar remained under French domination, first as a colony, then as a territoire d'outre-mer, finally as a member of the Communauté française. One of the most important elements of the « mission » of the colonizer was to assure the administration of justice. French policy consisted in introducing into the judiciary organization French judges whose task it was to bring to the country Western values, thus playing an important role in colonial domination. They were obliged, however, at the same time, to respect certain traditional native institutions. Their decisions also concerned the recently settled European population of Madagascar, for whom French metropolitan law had to be specially adapted.There were therefore three tasks to be accomplished by the judges of the Court of Appeals of Madagascar : to implement a special law to the French population, to implement their customary laws to the indigenous population and implement the law of metropolitan France, « eminent symbol of French culture », to that indigenous population. In practice, despite the difficulties encountered and the limits imposed on them, the judges managed these delicate tasks rather well. In confidently granting themselves all the liberty necessary to their function, the judges managed to satisfy the various communities in Madagascar, while remaining faithful to colonial policy.

In this thesis we consider solution approaches for the index tacking problem, in which we aim to reproduces the performance of a market index without purchasing all of the stocks that constitute the index. We solve the problem using three different solution approaches: Mixed Integer Programming (MIP), Genetic Algorithms (GAs), and Ant-colony Optimization (ACO) Algorithm by limiting the number of stocks that can be held. Each index is also assigned with different cardinalities to examine the change to the solution values. All of the solution approaches are tested by considering eight market indices. The smallest data set only consists of 31 stocks whereas the largest data set includes over 2000 stocks. The computational results from the MIP are used as the benchmark to measure the performance of the other solution approaches. The Computational results are presented for different solution approaches and conclusions are given. Finally, we implement post analysis and investigate the best tracking portfolios achieved from the three solution approaches. We summarise the findings of the investigation, and in turn, we further improve some of the algorithms. As the formulations of these problems are mixed-integer linear programs, we use the solver ‘Cplex’ to solve the problems. All of the programming is coded in AMPL.

Abstract The yeast Saccharomyces cerevisiae on solid media forms multicellular colonies. Cells within colonies undergo differentiation and metabolic diversification, including formation of two layers of cells called Upper and Lower cells. The metabolic activity of U and L cells is different. For instance a higher level of autophagy was observed in U cells. This thesis includes a literature review of molecular mechanisms of autophagy in yeasts. Yeast colonies, under starvation conditions, produce volatile ammonia signal. This signal allows them communicate over a distance. Studies, revealing cell differentiation within colonies and ammonia signalling among colonies, were performed in colonies of laboratory strains. Strains isolated from nature, so-called wild strains, form distinct structured colonies, termed fluffy. Yeast within fluffy colonies also form different cell types. However the situation seems to be more complex that within smooth colonies of laboratory strains. Strains were constructed during this study, which express marker proteins Icl2p, Pox1p, Mae1p, Pma1p, Pma2p, Ino1p, Met17p and Atg8p fused with fluorescent labels in order to study cell differentiation and other processes within fluffy colonies. Furthermore, a new system for ...