Готові списки джерел за темами / Optimisation de hit en lead

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Добірка наукової літератури з теми "Optimisation de hit en lead"

Автор: Grafiati
Опубліковано: 29 березня 2025

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Статті в журналах з теми "Optimisation de hit en lead"

1

Padalino, Gilda, Nelly El-Sakkary, Lawrence J. Liu, Chenxi Liu, Danielle S. G. Harte, Rachel E. Barnes, Edward Sayers, et al. "Anti-schistosomal activities of quinoxaline-containing compounds: From hit identification to lead optimisation." European Journal of Medicinal Chemistry 226 (December 2021): 113823. http://dx.doi.org/10.1016/j.ejmech.2021.113823.

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2

El Bakali, Jamal, Lucie Maingot, Julie Dumont, Helene Host, Akila Hocine, Nicolas Cousaert, Sandrine Dassonneville, Florence Leroux, Benoit Deprez, and Rebecca Deprez-Poulain. "Novel selective inhibitors of neutral endopeptidase: discovery by screening and hit-to-lead optimisation." MedChemComm 3, no. 4 (2012): 469. http://dx.doi.org/10.1039/c2md00287f.

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3

Bobrovs, Raitis, Jekaterina Bolsakova, Jhon Alexander Rodriguez Buitrago, Larisa Varaceva, Marija Skvorcova, Iveta Kanepe, Anastasija Rudnickiha, Emilio Parisini, and Aigars Jirgensons. "Structure-based identification of salicylic acid derivatives as malarial threonyl tRNA-synthetase inhibitors." PLOS ONE 19, no. 4 (April 1, 2024): e0296995. http://dx.doi.org/10.1371/journal.pone.0296995.

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Анотація:
Emerging resistance to existing antimalarial drugs drives the search for new antimalarials, and protein translation is a promising pathway to target. Threonyl t-RNA synthetase (ThrRS) is one of the enzymes involved in this pathway, and it has been validated as an anti-malarial drug target. Here, we present 9 structurally diverse low micromolar Plasmodium falciparum ThrRS inhibitors that were identified using high-throughput virtual screening (HTVS) and were verified in a FRET enzymatic assay. Salicylic acid-based compound (LE = 0.34) was selected as a most perspective hit and was subjected to hit-to-lead optimisation. A total of 146 hit analogues were synthesised or obtained from commercial vendors and were tested. Structure-activity relationship study was supported by the crystal structure of the complex of a salicylic acid analogue with a close homologue of the plasmodium target, E. coli ThrRS (EcThrRS). Despite the availability of structural information, the hit identified via virtual screening remained one of the most potent PfThrRS inhibitors within this series. However, the compounds presented herein provide novel scaffolds for ThrRS inhibitors, which could serve as starting points for further medicinal chemistry projects targeting ThrRSs or structurally similar enzymes.
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4

Mostarda, Serena, Tugçe Gür Maz, Alessandro Piccinno, Bruno Cerra, and Erden Banoglu. "Optimisation by Design of Experiment of Benzimidazol-2-One Synthesis under Flow Conditions." Molecules 24, no. 13 (July 3, 2019): 2447. http://dx.doi.org/10.3390/molecules24132447.

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Анотація:
A novel flow-based approach for the preparation of benzimidazol-2-one (1) scaffold by the 1,1′-carbonyldiimidazole (CDI)-promoted cyclocarbonylation of o-phenylenediamine (2) is reported. Starting from a preliminary batch screening, the model reaction was successfully translated under flow conditions and optimised by means of design of experiment (DoE). The method allowed the efficient preparation of this privileged scaffold and to set up a general protocol for the multigram-scale preparation in high yield, purity, and productivity, and was successfully applied for the multigram flow synthesis of N-(2-chlorobenzyl)-5-cyano-benzimidazol-2-one, which is a key synthon for hit-to-lead explorations in our anti-inflammatory drug discovery program.
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5

Osborne, James, Stanislava Panova, Magdalini Rapti, Tatsuya Urushima, and Harren Jhoti. "Fragments: where are we now?" Biochemical Society Transactions 48, no. 1 (January 27, 2020): 271–80. http://dx.doi.org/10.1042/bst20190694.

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Анотація:
Fragment-based drug discovery (FBDD) has become a mainstream technology for the identification of chemical hit matter in drug discovery programs. To date, the food and drug administration has approved four drugs, and over forty compounds are in clinical studies that can trace their origins to a fragment-based screen. The challenges associated with implementing an FBDD approach are many and diverse, ranging from the library design to developing methods for identifying weak affinity compounds. In this article, we give an overview of current progress in fragment library design, fragment to lead optimisation and on the advancement in techniques used for screening. Finally, we will comment on the future opportunities and challenges in this field.
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6

Taki, Aya C., Joseph J. Byrne, Tao Wang, Brad E. Sleebs, Nghi Nguyen, Ross S. Hall, Pasi K. Korhonen, et al. "High-Throughput Phenotypic Assay to Screen for Anthelmintic Activity on Haemonchus contortus." Pharmaceuticals 14, no. 7 (June 26, 2021): 616. http://dx.doi.org/10.3390/ph14070616.

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Анотація:
Parasitic worms cause very significant diseases in animals and humans worldwide, and their control is critical to enhance health, well-being and productivity. Due to widespread drug resistance in many parasitic worms of animals globally, there is a major, continuing demand for the discovery and development of anthelmintic drugs for use to control these worms. Here, we established a practical, cost-effective and semi-automated high throughput screening (HTS) assay, which relies on the measurement of motility of larvae of the barber’s pole worm (Haemonchus contortus) using infrared light-interference. Using this assay, we screened 80,500 small molecules and achieved a hit rate of 0.05%. We identified three small molecules that reproducibly inhibited larval motility and/or development (IC50 values of ~4 to 41 µM). Future work will critically assess the potential of selected hits as candidates for subsequent optimisation or repurposing against parasitic nematodes. This HTS assay has a major advantage over most previous assays in that it achieves a ≥ 10-times higher throughput (i.e., 10,000 compounds per week), and is thus suited to the screening of libraries of tens of thousands to hundreds of thousands of compounds for subsequent hit-to-lead optimisation or effective repurposing and development. The current assay should be adaptable to many socioeconomically important parasitic nematodes, including those that cause neglected tropical diseases (NTDs). This aspect is of relevance, given the goals of the World Health Organization (WHO) Roadmap for NTDs 2021–2030, to develop more effective drugs and drug combinations to improve patient outcomes and circumvent the ineffectiveness of some current anthelmintic drugs and possible drug resistance.
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7

Zaidi, Norburhanuddin Johari, Adib Afandi Abdullah, Choon Han Heh, Chun-Hung Lin, Rozana Othman, and Abdullah Al Hadi Ahmad Fuaad. "Hit-to-Lead Short Peptides against Dengue Type 2 Envelope Protein: Computational and Experimental Investigations." Molecules 27, no. 10 (May 18, 2022): 3233. http://dx.doi.org/10.3390/molecules27103233.

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Анотація:
Data from the World Health Organisation show that the global incidence of dengue infection has risen drastically, with an estimated 400 million cases of dengue infection occurring annually. Despite this worrying trend, there is still no therapeutic treatment available. Herein, we investigated short peptide fragments with a varying total number of amino acid residues (peptide fragments) from previously reported dengue virus type 2 (DENV2) peptide-based inhibitors, DN58wt (GDSYIIIGVEPGQLKENWFKKGSSIGQMF), DN58opt (TWWCFYFCRRHHPFWFFYRHN), DS36wt (LITVNPIVTEKDSPVNIEAE), and DS36opt (RHWEQFYFRRRERKFWLFFW), aided by in silico approaches: peptide–protein molecular docking and 100 ns of molecular dynamics (MD) simulation via molecular mechanics using Poisson–Boltzmann surface area (MMPBSA) and molecular mechanics generalised Born surface area (MMGBSA) methods. A library of 11,699 peptide fragments was generated, subjected to in silico calculation, and the candidates with the excellent binding affinity and shown to be stable in the DI-DIII binding pocket of DENV2 envelope (E) protein were determined. Selected peptides were synthesised using conventional Fmoc solid-phase peptide chemistry, purified by RP-HPLC, and characterised using LCMS. In vitro studies followed, to test for the peptides’ toxicity and efficacy in inhibiting the DENV2 growth cycle. Our studies identified the electrostatic interaction (from free energy calculation) to be the driving stabilising force for the E protein–peptide interactions. Five key E protein residues were also identified that had the most interactions with the peptides: (polar) LYS36, ASN37, and ARG350, and (nonpolar) LEU351 and VAL354; these residues might play crucial roles in the effective binding interactions. One of the peptide fragments, DN58opt_8-13 (PFWFFYRH), showed the best inhibitory activity, at about 63% DENV2 plague reduction, compared with no treatment. This correlates well with the in silico studies in which the peptide possessed the lowest binding energy (−9.0 kcal/mol) and was maintained steadily within the binding pocket of DENV2 E protein during the MD simulations. This study demonstrates the use of computational studies to expand research on lead optimisation of antiviral peptides, thus explaining the inhibitory potential of the designed peptides.
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8

Kandil, Sahar B., Samuel R. Jones, Sonia Smith, Stephen E. Hiscox, and Andrew D. Westwell. "Structure-Based Virtual Screening, Synthesis and Biological Evaluation of Potential FAK-FAT Domain Inhibitors for Treatment of Metastatic Cancer." Molecules 25, no. 15 (July 31, 2020): 3488. http://dx.doi.org/10.3390/molecules25153488.

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Анотація:
Focal adhesion kinase (FAK) is a tyrosine kinase that is overexpressed and activated in several advanced-stage solid cancers. In cancer cells, FAK promotes the progression and metastasis of tumours. In this study, we used structure-based virtual screening to filter a library of more than 210K compounds against the focal adhesion targeting FAK-focal adhesion targeting (FAT) domain to identify 25 virtual hit compounds which were screened in the invasive breast cancer line (MDA-MB-231). Most notably, compound I showed low micromolar antiproliferative activity, as well as antimigratory activity. Moreover, examination in a model of triple negative breast cancer (TNBC), revealed that, despite not effecting FAK phosphorylation, compound I significantly impairs proliferation whilst impairing focal adhesion growth and turnover leading to reduced migration. Further optimisation and synthesis of analogues of the lead compound I using a four-step synthetic procedure was performed, and analogues were assessed for their antiproliferative activity against three breast cancer (MDA-MB-231, T47D, BT474) cell lines and one pancreatic cancer (MIAPaCa2) cell line. Compound 5f was identified as a promising lead compound with IC50 values in the range of 4.59–5.28 μM in MDA-MB-231, T47D, BT474, and MIAPaCa2. Molecular modelling and pharmacokinetic studies provided more insight into the therapeutic features of this new series.
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9

Scanlon, Martin. "Monash Fragment Platform." Impact 2018, no. 3 (June 15, 2018): 32–34. http://dx.doi.org/10.21820/23987073.2018.3.32.

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Анотація:
The Monash Fragment Platform (MFP) provides academic and commercial researchers with access to fragment-based drug discovery (FBDD) technologies for their therapeutic targets. The facilities are located at the Monash Institute for Pharmaceutical Sciences (MIPS) in Parkville, Australia's largest and most successful pharmaceutical institute. FBDD screening uses our high-quality, in-house fragment library designed to maximise chemical space coverage and enable rapid hit optimisation. Screening cascades are tailored for each target and can be performed using a variety of techniques including NMR spectroscopy and our state-of-the-art surface plasmon resonance (SPR) facility. Structural biology and medicinal chemistry expertise utilising our standardized REFIL strategy enables the rapid elaboration of fragments into leads without the requirement for a large chemistry program.
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10

Mager, Magnus. "Noise performance of the ALPIDE-based ALICE Inner Tracking System." Journal of Physics: Conference Series 2374, no. 1 (November 1, 2022): 012062. http://dx.doi.org/10.1088/1742-6596/2374/1/012062.

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Анотація:
The new ALICE Inner Tracking System is the first large-scale MAPS-based tracking system. It is covering an active surface of 10 m2, with a total of 12.5 billion pixels. Several optimisations of the pixel chip lead to a quasi-noise-free operation, with measured fake-hit rates of below 1 hit per pixel and billion events (system level numbers), numbers compatible with the order of magnitude expected from cosmic and natural background radiation. This contribution covers a detailed study of fake hits as recorded in the inner-most detector barrels, made of the highest quality chips. It reveals a localised noise pattern that could be traced down to originate from decoupling capacitors present on the detector module. It can most likely be explained by the radioactive decay of 210Pb, which is present in trace amounts in the solder that was used to mount decoupling capacitors on the detector modules. This hypothesis is substantiated with a dedicated simulation study and laboratory measurements.
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Більше джерел

Дисертації з теми "Optimisation de hit en lead"

1

Quevedo, Camilo E. "Design and synthesis of Quinazolinone-based libraries for inhibitation of Kinase activity and hit-to-lead optimisation of Wnt pathway inhibitors." Thesis, Institute of Cancer Research (University Of London), 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.510367.

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2

Borzakian, Sibyline. "Identification et développement chimique d'inducteurs de protéines immunorégulatrices (SLPI et IL-10) à visée thérapeutique." Electronic Thesis or Diss., université Paris-Saclay, 2024. http://www.theses.fr/2024UPASF012.

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Анотація:
Les travaux réalisés durant cette thèse ont été scindés en trois parties distinctes, s'inscrivant toutes dans le cadre du développement de composés à visée thérapeutique. La première partie de ce manuscrit est consacrée à l'identification d'inducteurs de SLPI (Secretory Leucocyte Protease Inhibitor), une protéine principalement produite par les cellules immunitaires et épithéliales, et possédant de nombreuses propriétés biologiques prometteuses. Un criblage phénotypique a donc été mis en œuvre à partir de deux chimiothèques académiques et a mené à l'identification de 5 hits, capables d'induire la production de SLPI par les lymphocytes B. Ces travaux ont donc permis d'ouvrir la voie aux applications thérapeutiques potentielles de ces composés, notamment dans le cadre du développement de traitements antibiotiques ou anti-inflammatoires. La deuxième partie de ce manuscrit est consacrée à l'identification d'un pharmacophore d'inducteurs d'Interleukine-10 (IL-10), une cytokine produite par les cellules immunitaires, et ayant pour rôle principal la régulation de l'immunité. Préliminairement à ces travaux de thèse, un criblage phénotypique a permis d'identifier des inducteurs de la production d'IL-10 par les lymphocytes B à des fins de thérapie cellulaire visant à réduire les symptômes de la sclérose en plaque. Les travaux réalisés dans le cadre de cette thèse ont donc consisté en l'optimisation hit-to-lead du composé le plus prometteur, afin d'identifier un pharmacophore d'inducteurs d'IL-10, via l'établissement de relations structure-activité robustes. Durant ces travaux, 240 analogues ont été synthétisés. Enfin, la troisième partie de ce manuscrit est consacrée au développement d'une méthodologie de synthèse par voie photochimique, pour l'accès à des composés hétérocycliques originaux. Cette méthodologie ne nécessitant ni chauffage, ni catalyseur métallique, s'inscrit donc pleinement dans le développement d'une chimie plus verte
The work carried out during this thesis has been divided into three distincts parts, all relating to the developpement of compounds for therapeutic use. The first part of this manuscript is devoted to the identification of inducers of Secretory Leucocyte Protease Inhibitor (SLPI), a protein mainly produced by immune and epithelial cells, with numerous promising biological properties. A phenotypic screening was conducted using two academic chemical libraries, leading to the identification of 5 hits able of inducing SLPI production by B cells. This work paved the way for potential therapeutic applications of these compounds, particularly in the development of antibiotic or anti-inflammatory treatments. The second part of this manuscript is dedicated to identifying a pharmacophore for inducers of Interleukin-10 (IL-10), a cytokine produced by immune cells, whose main role is the regulation of immunity. Prior to this thesis, a phenotypic screening was conducted to identifiy inducers of IL-10 production by B cells for use in cell therapy to reduce symptoms of multiple sclerosis. This thesis work involved hit-to-lead optimization of the most promising compound to identify a pharmacophore for IL-10 inducers through the establishment of robust structure-activity relationships. During this phase, 240 analogs were synthetized. Finally, the third part of this manuscript focuses on the development of a photochemical synthesis methology for accessing original heterocylic compounds. This methology, requiring neither heating nor metallic catalysts, aligns with the development of greener chemistry
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3

Samson, Samantha. "Profilage in silico de la protéine multifonctionnelle Mfd, une cible thérapeutique innovante dans la lutte contre l'antibiorésistance bactérienne." Electronic Thesis or Diss., université Paris-Saclay, 2024. http://www.theses.fr/2024UPASL125.

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Анотація:
Face à la montée préoccupante de la résistance aux antibiotiques, la recherche de nouveaux antimicrobiens constitue un enjeu de santé urgent. Nous avons identifié la protéine bactérienne Mutation Frequency Decline (Mfd) comme étant une cible thérapeutique innovante pour le développement de nouveaux médicaments. Un criblage in silico à haut débit a été réalisé, dans un premier temps, afin de sélectionner des molécules pouvant se lier spécifiquement au site actif de la protéine et inhiber son activité, empêchant donc la résistance bactérienne face au stress immunitaire de l'hôte. Les molécules identifiées se sont révélées efficaces in vitro et efficaces et non toxiques in vivo, dans un modèle d'infection chez l'insecte, sur au moins deux pathogènes bactériens. Ces données préliminaires ont constitué une preuve de concept du potentiel innovant de ces molécules et ont également servi de base aux travaux présentés dans cette thèse.L'objectif principal de ce travail était l'identification des interactions moléculaires critiques entre ces molécules inhibitrices et le site actif de Mfd chez E. coli, ainsi que l'élargissement de cette analyse aux pathogènes prioritaires du groupe ESKAPE. En conséquence de quoi, un modèle optimal d'inhibiteur a été déterminé et ses dérivés sont actuellement testés in vitro et in vivo en tant que potentiels agents antimicrobiens. En parallèle, l'analyse de la séquence et de la structure de Mfd, provenant de souches environnementales et cliniques, met en évidence les caractéristiques d'une corrélation moléculaire entre la séquence de Mfd et le phénotype de virulence du pathogène. La confirmation in vitro est actuellement en cours d'évaluation. Enfin, mon objectif final est de repositionner la fonction motrice de Mfd dans un cadre plus large de remodelage conformationnel et fonctionnel afin de mieux comprendre cette cible et son rôle dans la réparation par excision de nucléotides. En produisant plusieurs simulations de dynamique moléculaire sur des « linkers » clés qui relient les principaux modules fonctionnels de Mfd, l'investigation de leur flexibilité intrinsèque et de leur conservation vise à récapituler le remodelage extensif des conformations de Mfd au cours de son cycle fonctionnel, tel qu'il a été décrit précédemment par cryo-EM. Cela a pour but de documenter dans quelle mesure ces linkers, qui relient cette protéine multi-modulaire, sont plus que de simples "connecteurs" et portent, dans leur séquence et leur longueur, des propriétés internes leur permettant d'adopter des états discrets garantissant la transition boucle-hélice nécessaire au bon fonctionnement de Mfd
In an alarming context of antibiotic resistance, the search for new antimicrobials is an urgent health issue. We had identified the bacterial protein Mutation Frequency Decline (Mfd) as an innovative target for the development of new drugs. A high throughput in silico screening was initially performed in order to select molecules specifically binding to the active site of the target and inhibiting its activity, thereby preventing bacterial resistance to host immune stress. The identified hits were shown to be efficient in vitro and efficient and non-toxic in vivo, in an insect model of infection on at least two bacterial pathogens. These preliminary data have constituted a proof of concept of the innovative potential of these hits and were also the basis of this thesis.The main objective of this work was the identification of the critical molecular interaction found between those hits and the active site of Mfd in E. coli and also enlarged to the priority pathogens of the ESKAPE group. As a result, an optimal inhibitor scaffold was determined and its derivatives are currently tested in vitro and in vivo as potential antimicrobial agents. In parallel, the sequence and structure analysis of Mfd, from environmental and clinical strains, showcase the basic features of a molecular correlation between Mfd sequence and virulence phenotype. The in vitro confirmation is currently being evaluated. Finally, my goal reposition this motor function of Mfd into a larger conformational and functional remodeling of Mfd in order to get a better understanding of this target and its role in the Nucleotide Excision Repair. Using molecular dynamics simulation on distinguished linkers that connect the main functional modules of Mfd, the investigation of their intrinsic flexibility and resilience to recapitulate the extensive remodeling of Mfd conformations within its functional cycle that has been previously described by cryo-EM. This aims to document to which extent the linkers that connect this multi-module protein are more than "linkers" and harbor, in their sequence and length, internal properties to adopt discrete states that guarantee disorder-to-coil transition to assure the functional machinery of Mfd
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4

Papadatos, George. "Data mining for lead optimisation." Thesis, University of Sheffield, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.556989.

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Анотація:
The recurring theme of this thesis is the application of diverse data mining and chemoinformatics techniques to structural and experimental property data, and particularly to data produced during the stage of drug discovery called lead optimisation. The work reported here seeks to provide more than one rational answer to the real-life issues routinely facing medicinal chemists. The thesis is divided into three parts: In the first part, several methodologies are described which facilitate the automatic mining of temporal, hierarchical lead optimisation data from the archives. Then, these data are appropriately used to provide informative visualisations, with regard to the exploration of chemical space, both locally (i.e. on a chemical array level) and globally (i.e. in the whole project). Finally, several ways of assessing the progress of a particular lead optimisation project are investigated. The second part of the thesis compares and assesses the relative merits of two computational methods that quantify the neighbourhood behaviour of a descriptor. The main conclusions of this part are two-fold: firstly, the optimality criterion method is demonstrated to be a suitable way to select descriptors for the systematic exploration of chemical space during array-based lead optimisation; secondly, regarding the actual neighbourhood behaviour performance exhibited by twelve types of fingerprints, it is shown that circular-based ones perform consistently better than the others and, notably, at a much lower similarity threshold. The third part focuses on explicit structural transformations between molecular pairs and their impact on properties such as hERG channel blocking, solubility and lipophilicity. More importantly, the study investigates the context of a transformation and its role on the impact of a particular modification. Using substructural descriptors to represent the context of a transformation, and considering both the local and the global environment, several contextsensitive cases are identified and rationalised. Overall, it is demonstrated that the inclusion of contextual information can enhance the predictive power of matched molecular pair analysis. Several context-sensitive examples are also identified in publicly available data.
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5

Neiström, Linda. "Characterisation of Used Lead-Acid Batteries for Feed Optimisation in Secondary Lead Production." Thesis, Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:ltu:diva-70740.

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Анотація:
Used lead-acid batteries are the main raw material in secondary lead production. Technologicalprogress in the car industry, have raised concerns regarding the lead-acid battery’s leading positionas electrochemical storage system in the future. However, the lead-acid battery industry isadvancing with innovations, such as hybrid and micro-hybrid vehicles, and is believed to have acontinued important role in the rechargeable battery market. Lead is one of the most investigatedmetal due to concerns from society regarding the negative effect on human health and theenvironment. Consequently, that has led to tighter controls and regulations of the lead processingindustry which, in turn, has led to technological improvement concerning design and operation ofthe lead processing plants. Used lead-acid batteries have a complex composition with a variety ofcomponents made of lead (i.e., metallic, oxide or sulphate) and non-lead materials (plastics andelectrolyte). Traditionally, battery recycling is done without separating those components.However, to optimise the use of resources and decrease the environmental impact of secondarylead production, a pre-treatment step to the batteries recycling is desired. The pre-treatment iscomprised of breaking the batteries and separation of the battery components. The aim of the present thesis, was to study the components of a used lead-acid battery, which willbe the outgoing material of a future plastic separation plant at Boliden Bergsöe, and their effect onthe process. Furthermore, the study aimed at investigating the possibilities to adjust the feedcomposition for further process optimisation and improvement of the process quality in terms ofenergy usages and environmental impact. This was done by characterise, through qualitative andquantitative composition and mass distribution, the fractions in a used lead-acid car battery. Four lead-acid car batteries were provided for dismantling to study mass distribution, and toliberate the components for further analysis. The analytical techniques used in this study werequalitative and quantitative (Rietveld) XRD analysis, SEM-EDS and TGA. The result showed thatoverestimated amount of battery separators (PE) has been used at production planning in energyandemission calculations, which can cause financial losses due to overpaid emission tax. The pastefraction showed a large variation in mass between the studied batteries and consists mostly ofPbSO4. Large variation in the paste mass may cause uneven sulphur emission from smelter.Consequently, this reinforces the need for implementation of the separation of the battery feed fora better control of the paste addition to the smelter. When PE decomposes in the shaft furnace the remaining ash will mainly consist of silica, whichwill affect the sulphur uptake in the shaft furnace. The large content of silica leads to a lower energycontribution to the process; however, a lower content of hydrocarbons leads to lower CO2emissions. If a desulphurisation of the pastes would be implemented, it is believed to affect theprocess through a decreased need of coke and iron. A reduced usage of coke and iron would lowerthe production costs and lead to decreased CO2 emissions. A desulphurisation will also decreasethe lead sulphate content in the feed, thus lead to better control of the sulphur emissions. This study provides additional support and further insight into composition and mass distributionof the components in a lead-acid battery. Furthermore, the study indicates possible impact of thefuture separation on the new feed properties and on the subsequent processing.
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6

Yglesias, Tatiana. "Application of process analysis and optimization tools in hit-to-lead and lead optimization phases of drug discovery at EPP, NIBR." Thesis, Massachusetts Institute of Technology, 2010. http://hdl.handle.net/1721.1/59882.

Повний текст джерела
Анотація:
Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Chemical Engineering; and, (M.B.A.)--Massachusetts Institute of Technology, Sloan School of Management; in conjunction with the Leaders for Global Operations Program at MIT, 2010.
Cataloged from PDF version of thesis.
Includes bibliographical references (p. 82-84).
Given that research is based on innovation, it has been believed that its activities can only be optimized with equipment upgrade, increment in personnel scientific knowledge, development of new analytical software and/or changing the areas of study. After realizing the limited results achieved with these approaches, lab representatives started to notice the opportunity of introducing process optimization tools, such as Lean and Six Sigma, which showed success in manufacturing environments,. This project analyzes the interrelation between process and results, providing a clear explanation of cause and effect conditions, and a concise list of areas for improvement. Specifically, the document defines a measurement system using process maps and key performance indicators (KPIs). With this, the document describes the current state through historic trends, provides a complete data and root cause analysis for current state description, and provides a process capability study for the available indicators. Implementation of the steps mentioned above show how focus in lab turnaround times have been deviating attention from more impactful improvements, which can greatly affect overall drug discovery duration. Also, the analysis identifies that constant technology changes caused constant adaptation of process procedures, which generated non-value added activities. These non-value added activities today occupy about 50% of a lab associate's time. Lastly, historic data evaluation shows that root cause statistical analysis is limited by the presence of a combination of special and common cause variations. Some of the project recommendations include: incorporation of chemist's knowledge about compound potency, integration of equipment and software information, change in booking system, incorporation of assay and plate criteria, definition of standard procedures for specific activities, and integration of assay development and data submission tools. Overall, these changes can lead to a 50% reduction in the profiling times greater than 60 days, decrease of 62% and 60% in Compound Manager (CM) and Compound Profiler (CP) non-value added times respectively, 30% decrease in CM and CP total duration per assay plate, and increase in profiling time stability and predictability. Despite the fact that timing and scale of available resources will impact the realized benefits, the proposed framework gives EPP the opportunity to assess the improvements by their effect and alignment with goals.
by Tatiana Yglesias.
M.B.A.
S.M.
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Voitovich, Iuliia. "Les inhibiteurs d'interaction protéine-protéine, une stratégie innovante en cancérologie." Thesis, Aix-Marseille, 2018. http://www.theses.fr/2018AIXM0701.

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Les protéines BET, modules impliqués dans la régulation épigénétique, jouent un rôle essentiel dans le développement du cancer. Actuellement plusieurs inhibiteurs de protéines BET font l'objet d'essais cliniques pour le traitement de différents types de cancer. Une des limitations au développement clinique est l'impact général de la modulation génique induite par des inhibiteurs de type ‘pan-BET’ non sélectifs. Un ciblage individuel des protéines BET et la discrimination des domaines BD1 et BD2 conduiraient à un effet transcriptionnel plus spécifique limitant les effets secondaires et l’apparition de résistance. Le criblage d’une chimiothèque focalisée sur les interactions protéine-protéine a permis d’identifier deux molécules avec des profils de sélectivité uniques. Une étude de SAR a révélé le fragment minimal nécessaire à l’interaction ligand-protéine. La résolution de la structure cristallographique en complexe avec Brd4(BD1) a permis de valider nos interprétations et de développer des inhibiteurs BET plus puissants et sélectifs. Une stratégie de synthèse originale orientée vers une diversité structurale (DOTS) combinant le criblage virtuel et l’élaboration automatisée de bibliothèques focalisées a été utilisée. Ce travail a découvert un inhibiteur optimisé de BD1 avec une affinité de 100x supérieure à la molécule initiale et un ratio de sélectivité BD1 vs BD2 égale à 300. L’activité cellulaire d’inhibition du pro-oncogène c-Myc, au µM, a permis de valider le composé en tant que sonde moléculaire. Des études in vitro et in vivo permettront d'élucider le rôle biologique individuel de chaque BD et de valider l'intérêt de leurs développement en clinique
BET-proteins, acting as epigenetic readers, play an essential role in cancer development. To date, numerous potent inhibitors disrupting BET functions have been discovered, including several of them that are undergoing clinical trials for the treatment of different types of cancer. The common drawback limiting their use in clinical practice is an inability to distinguish between BET-members that may cause side effects and resistances. The selective targeting of individual BET and the discrimination between BD1 and BD2 present an opportunity to achieve more selective transcriptional effect. A midthroughput screening of previously designed chemical library allowed identification of two molecules with unique profiles of selectivity that have never been observed. An undertaken structure-based program revealed a minimum scaffolds necessary for binding. Taking together with resolved X-Ray structures it allowed the development of more potent and selective BET inhibitors by DOTS (diversity oriented target focused synthesis) strategy, combining virtual screening and diversity oriented library design. This optimization led to a potent inhibitor with up to 100-fold improvement of affinity to the target and up to 300-fold selectivity toward BD1. Dose-response downregulation of c-Myc levels in low micromolar range in cell assays allowed the validation of the identified molecule as a chemical probe. Further comprehensive in vitro and in vivo evaluations of this compound will enable elucidating the biological role of each bromodomain and a validation of the interest toward the development of selective inhibitors in clinic
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Mariani, Alessandro. "Optimisation of valve regulated lead acid battery design for high power applications." Thesis, University of South Wales, 2017. https://pure.southwales.ac.uk/en/studentthesis/optimisation-of-valve-regulated-lead-acid-battery-design-for-high-power-applications(f8316110-ebc1-4f0b-9e1b-4a66d7e50564).html.

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Filippi, Andrea <1990&gt. "An application of Genetic Algorithms for the Lead Molecule Optimisation. (Titolo provvisorio)." Master's Degree Thesis, Università Ca' Foscari Venezia, 2016. http://hdl.handle.net/10579/9366.

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Lead Molecule Optimisation and the process of Drug Design are very difficult problems, demanding an huge investment in terms of resources and experimentation. The standard approach requires iterations of long synthesis and testing cycles as well as formulations, which are hard and complex tasks. In this thesis are presented, in the first chapters, an analysis of Nature inspired computation and the problem of Lead Molecule Optimisation; lately it is proposed a method to address such problem. It is the Genetic Algorithm Optimisation (GAO) method, which will be applied to a set of experimental data of fitness values and molecule fragments. The dataset is provided by the European Center for Living Technology (ECLT). The GAO is used to find an optimal solution for the development of a particular drug (MMP-12 inhibitors). A total of 120 molecules are tested over 6 generations; the results show that the algorithm works as intended by reaching the optimum in an acceptable amount of time, required for the computation. The good performance of the GAO method is analysed and shown in the thesis with its capacity to reach the desired optimum value using the data set provided.
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Moffat, Kirstin. "Development of computational methods for 3D similarity and structure-based design techniques in lead optimisation." Thesis, University of Sheffield, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.434521.

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Книги з теми "Optimisation de hit en lead"

1

Faller, Bernard, and Laszlo Urban, eds. Hit and Lead Profiling. Wiley, 2009. http://dx.doi.org/10.1002/9783527627448.

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Ramaswamy, Vijayan, and Vasanthanathan Poongavanam. Computational Drug Discovery: From Hit to Lead Optimization. Wiley & Sons, Limited, John, 2023.

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Optimisation of Corrosion Control for Lead in Drinking Water Using Computational Modelling Techniques. IWA Publishing, 2013.

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Hayes, Colin, Croft, T N. Optimisation of Corrosion Control for Lead in Drinking Water Using Computational Modelling Techniques. IWA Publishing, 2012.

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Urban, Laszlo, Gerd Folkers, Raimund Mannhold, Hugo Kubinyi, and Bernard Faller. Hit and Lead Profiling: Identification and Optimization of Drug-Like Molecules. Wiley & Sons, Limited, John, 2010.

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Urban, Laszlo, Gerd Folkers, Raimund Mannhold, Hugo Kubinyi, and Bernard Faller. Hit and Lead Profiling: Identification and Optimization of Drug-Like Molecules. Wiley & Sons, Limited, John, 2010.

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Urban, Laszlo, Gerd Folkers, Raimund Mannhold, Hugo Kubinyi, and Bernard Faller. Hit and Lead Profiling: Identification and Optimization of Drug-Like Molecules. Wiley & Sons, Limited, John, 2010.

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8

Hit and lead profiling: Identification and optimization of drug-like molecules. Weinheim: Wiley-VCH, 2009.

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9

Urban, Laszlo, Gerd Folkers, Raimund Mannhold, Hugo Kubinyi, and Bernard Faller. Hit and Lead Profiling: Identification and Optimization of Drug-Like Molecules. Wiley & Sons, Incorporated, John, 2009.

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10

Now Lead the Change: Repurpose Your Career, Future-Proof Your Organization, and Regenerate Our Crisis-Hit World by Mastering Transformational Leadership. Switch On Books, 2020.

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Частини книг з теми "Optimisation de hit en lead"

1

Mazanetz, Michael, Richard Law, and Mark Whittaker. "Hit and Lead Identification from Fragments." In De novo Molecular Design, 143–200. Weinheim, Germany: Wiley-VCH Verlag GmbH & Co. KGaA, 2013. http://dx.doi.org/10.1002/9783527677016.ch6.

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2

Dunlap, Norma, and Donna M. Huryn. "Drug discovery: hit and lead identification." In Medicinal Chemistry, 19–50. New York, NY : Garland Science, Taylor & Francis Group, LLC, [2018]: Garland Science, 2018. http://dx.doi.org/10.1201/9781315100470-2.

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3

Braggio, Simone, Mauro Corsi, Aldo Feriani, Stefano Fontana, Luciana Marocchio, and Caterina Virginio. "CHAPTER 15. Discovery Toxicology In Lead Optimisation." In The Handbook of Medicinal Chemistry, 364–412. Cambridge: Royal Society of Chemistry, 2015. http://dx.doi.org/10.1039/9781782621836-00364.

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4

Cregan, Vincent, and William T. Lee. "Optimisation of a Lead Sulphate Settling Process." In Trends in Mathematics, 1–6. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-030-01153-6_1.

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Connolly, Stephen, and Simon E. Ward. "CHAPTER 21. Lead Optimisation: What You Should Know!" In The Handbook of Medicinal Chemistry, 529–65. Cambridge: Royal Society of Chemistry, 2015. http://dx.doi.org/10.1039/9781782621836-00529.

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6

Kapoulitsas, P., M. Giunti, R. Hampson, A. Cranley, S. Gray, B. Kretschmer, R. Knight, and J. Clark. "Commissioning and Optimisation of the New Lead and Silver Refinery at the Pasminco Port Pirie Smelter." In Lead-Zinc 2000, 187–202. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2013. http://dx.doi.org/10.1002/9781118805558.ch10.

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Sewing, A., and D. Cawkill. "High-Throughput Lead Finding and Optimisation for GPCR Targets." In Ernst Schering Foundation Symposium Proceedings, 249–66. Berlin, Heidelberg: Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/2789_2006_012.

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8

Pignol, Jean-Philippe, Philippe Paquis, Roland Sabattier, Arlene Lennox, Thomas Kroc, Dan Jones, Jakobus Slabbert, Cheikh M. B. Diop, Gilbert Blondiau, and Noël Breteau. "BNCEFN Optimisation with Lead Blocks Collimation and Graphite Embedding." In Frontiers in Neutron Capture Therapy, 741–46. Boston, MA: Springer US, 2001. http://dx.doi.org/10.1007/978-1-4615-1285-1_107.

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Heifetz, Alexander, Michelle Southey, Inaki Morao, Andrea Townsend-Nicholson, and Mike J. Bodkin. "Computational Methods Used in Hit-to-Lead and Lead Optimization Stages of Structure-Based Drug Discovery." In Methods in Molecular Biology, 375–94. New York, NY: Springer New York, 2017. http://dx.doi.org/10.1007/978-1-4939-7465-8_19.

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Lipinski, Christopher A. "Overview of Hit to Lead: The Medicinal Chemist's Role from HTS Retest to Lead Optimization Hand Off." In Topics in Medicinal Chemistry, 1–24. Berlin, Heidelberg: Springer Berlin Heidelberg, 2009. http://dx.doi.org/10.1007/7355_2009_4.

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Тези доповідей конференцій з теми "Optimisation de hit en lead"

1

Aimetta, A., N. Abrate, M. Massone, and S. Dulla. "Advancements in the Genetic-Driven Optimisation of the Few-Group Structures for Lead Fast Reactors Analysis." In International Conference on Physics of Reactors (PHYSOR 2024), 1703–12. Illinois: American Nuclear Society, 2024. http://dx.doi.org/10.13182/physor24-43878.

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Mo¨ller, Bjo¨rn Fredriksson, Mitsuru Obana, Mohsen Assadi, and Athanasios Mitakakis. "Optimisation of HAT-Cycles – With and Without CO2 Capture." In ASME Turbo Expo 2004: Power for Land, Sea, and Air. ASMEDC, 2004. http://dx.doi.org/10.1115/gt2004-53734.

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In a world where distributed power generation and deregulation of energy markets are on everyone’s agenda, the need for highly efficient power plants with short lead times is greater than ever. Although at present combined cycles provide a solution, development of ever more advanced machines to increase efficiency and lower the environmental impact has led to high maintenance costs and a decrease in availability. The Humid Air Turbine (HAT) represents a different approach, suitable for distributed power generation in the medium power range. The HAT cycle, and other wet gas turbine cycles, which have been extensively studied during the last ten years, show as high an efficiency as that of combined cycles, but at a lower specific cost and, with inherently low emissions of NOx. Despite all research done no full-scale plant has been built as yet. CO2 capture is another concept widely studied in recent years. In the present study three HAT cycle configurations are investigated, two of them connected to a post-combustion CO2-capture plant. Thermodynamic and thermoeconomic optimisation of the plants was performed using genetic algorithms (GA), a robust optimisation technique based on Darwinian evolution theories. The three configurations studied were 1) a standard inter-cooled HAT cycle, referred to as the reference cycle, 2) the reference cycle together with an integrated CO2-capture plant taking the energy needed for the CO2 separation from the exhaust heat of the turbine, and 3) the reference cycle together with a CO2 capture plant, in which the energy is supplied by a separate bio-fuelled boiler. This third configuration enables all fossil-based CO2 to be separated. All power cycles were modelled using IPSEpro, a heat- and mass-balance software, employing advanced component models developed by the authors. It has an interface for optimisation and the possibility of employing user-defined objective functions. The impact of CO2 taxation was studied to determine showing which configuration is the most economical at the current fuel-price and tax-level.
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Pietrikova, Alena, L'ubomir Livovsky, Jan Urbancik, and Radoslav Bucko. "Optimisation of Lead Free Solders Reflow Profile." In 2006 29th International Spring Seminar on Electronics Technology. IEEE, 2006. http://dx.doi.org/10.1109/isse.2006.365149.

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4

"Searching for bioactive drug lead compounds from natural humic substances." In Seventh International Conference on Humic Innovative Technologies "Humic substances and technologies for resilience" (HIT – 2022). NP CBR "Humus Sapiens", 2022. http://dx.doi.org/10.36291/hit.2022.071.

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5

Alexander, Brian G., Roberta Pireddu, Nan N. Sun, Huidong Yu, Kara D. Forinash, Matthew P. Martin, Ernst Schonbrunn, Wayne C. Guida, Said M. Sebti, and Nicholas J. Lawrence. "Abstract 3243: Discovery of novel Rho kinase inhibitors: Hit generation and lead optimization." In Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL. American Association for Cancer Research, 2011. http://dx.doi.org/10.1158/1538-7445.am2011-3243.

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6

Seufitelli, Danilo Boechat, and Mirella M. Moro. "Understanding Musical Success Beyond Hit Songs: Characterization and Analyses of Musical Careers." In Anais Estendidos do Simpósio Brasileiro de Sistemas Multimídia e Web, 23–24. Sociedade Brasileira de Computação - SBC, 2024. http://dx.doi.org/10.5753/webmedia_estendido.2024.244326.

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Streaming brought the high data availability over the web associated with music consumption and listener preference. With such data, we can extract relevant knowledge, such as what can lead some songs to success and others not. In this scenario, Hit Song Science emerged, an area of study focused on revealing the dynamics of success in the music industry. Collecting hits can lead artists to experience periods of success far beyond the "ordinary" periods known as Hot Streaks. In this sense, understanding the factors of how the different profiles of artists stand out and reach their most successful periods can be crucial for the music industry, which deals with the constant natural evolution of the market and needs to reinvent itself to satisfy the desires of its consumers: connect successful music and artists. Thus, this thesis aims to identify the characteristics that lead artists to reach their most successful periods.
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7

Pireddu, Roberta, Harshani R. Lawrence, Mathew P. Martin, Stephane Betzi, Richard Yip, Hua Yang, Nan N. Sun, et al. "Abstract 3253: Novel oxindole inhibitors of Aurora A kinase: Structure based hit-to-lead approach." In Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL. American Association for Cancer Research, 2011. http://dx.doi.org/10.1158/1538-7445.am2011-3253.

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8

Kavicka, Frantisek, Josef Stetina, Karel Stransky, Jana Dobrovska, Bohumil Sekanina, and Jaromir Heger. "A Numerical Model of Solidification of a Massive Casting From Malleable Cast-Iron." In ASME 2004 Heat Transfer/Fluids Engineering Summer Conference. ASMEDC, 2004. http://dx.doi.org/10.1115/ht-fed2004-56043.

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A numerical model of the temperature field associated with solidifying castings aims to achieve two general goals: directed solidification (as the primary condition for a healthy casting), and optimisation of the technology of casting together with the preservation of optimum utility properties of the product. A specific goal of this model is the selection and optimisation of the method of cooling to shorten the solidification time to obtain a spherical graphite structure with good nodular properties, and with a sufficient density of graphite spheres (cells). The speed of cooling during solidification and cooling in the mould is therefore a significant quantity influencing the formation of the structure. The achievement of these goals depends on the ability to analyse and, successively, to control the effect of the main factors which characterise the solidification process or accompany it. The analysis of the quantities is focused on determining the causes of the formation of the heterogenic temperature field during casting, considering the phase and structural changes. It is also focused on the thermokinetics of the formation of shrinkage porosities and cavities and on the prediction of their formation. This leads to the optimisation of the shape and sizes of the risers, the method of insulation, the treatment of the level. The model is applicable to various shapes of castings. The software is capable of analysing the temperature field of the actual casting, as well as the temperature field of the mould and cores, including the dependence of their material. It is also capable of considering non-linearity, i.e. the dependence of the thermophysical properties—namely the material of the casting and mould on the temperature, as well as the dependence of the heat transfer coefficients on the surface and interface temperature. The model is also equipped with an original network generator (pre-processing) as well as graphical output (post-processing).
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Scherff, Maximilian L. D., Stefan Schwertheim, Yue Ma, Thomas Mueller, and Wolfgang R. Fahrner. "10 × 10 cm2 Hit Solar Cells Contacted with Lead-Free Electrical Conductive Adhesives to Solar Cell Interconnectors." In Conference Record of the 2006 IEEE 4th World Conference on Photovoltaic Energy Conversion. IEEE, 2006. http://dx.doi.org/10.1109/wcpec.2006.279709.

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Baines, Olivia, Rina Sha, Christopher O’Shea, and Davor Pavlovic. "BS10 Cardiac pacing location- optimisation of optical mapping protocols and pacemaker lead position." In British Cardiovascular Society Annual Conference, ‘Back to the patient’, 3–5 June 2024. BMJ Publishing Group Ltd and British Cardiovascular Society, 2024. http://dx.doi.org/10.1136/heartjnl-2024-bcs.236.

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Звіти організацій з теми "Optimisation de hit en lead"

1

Chhoeung, Norinmony, Sesokunthideth Chrea, and Nghia Nguyen. Cambodia’s Cash Transfer Program during COVID-19. Asian Development Bank Institute, December 2022. http://dx.doi.org/10.56506/rrmz8095.

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In 2019, Cambodia had been enjoying its steady economic growth until the coronavirus disease (COVID-19) pandemic hit the country from February 2020 in the form of severe infectious diseases, causing both economic and social problems for people from all walks of life, especially poor and vulnerable families. The IDPoor Equity Card, a poverty identification and registration system, was introduced in Cambodia to provide cash to poor pregnant women and children since 2016. Given its continued success, Cambodian Prime Minister Samdech Hun Sen announced the implementation of the Cash Transfer Program (CTP) using the IDPoor Card system. The CTP provided cash to poor and vulnerable households across the country affected by the pandemic. Executing the first large-scale cash transfer program in history presented significant challenges for the Government of Cambodia. In addition to implementing the program, which had to adhere to the three principles of equity–equality–efficiency, the government had other challenges to overcome, such as the limited number of tablets and facilities to accommodate the many people waiting in line to withdraw cash. Under the guidance of the central government, particularly the Economic and Finance Policy Committee, a technical working group was established to lead the implementation process; coordination among local governments, local councils, agencies, and the poor and needy; review of the IDPoor database; establishment of the digital payment system; and training of local government staff. This enabled the CTP to effectively distribute cash to poor and vulnerable households during the pandemic.
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Amzallag, Robert. The return of inflation: a banker’s perspective. CIRANO, May 2022. http://dx.doi.org/10.54932/egsn1582.

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The COVID 19 pandemic followed by the invasion of Ukraine is a two-punch economic strike never seen in recent history. The pandemic not only disrupted many aspects of a tightly knit integrated world but also exposed its fragility. The devastation of Ukraine and the vast program of sanctions quickly imposed by most major developed countries have accelerated the retreat of globalization. For many decades, Central Bankers and economists considered stable prices as an almost permanent feature. Today, the consumer price index in the US hit a 40-year high at more than 8 per cent and experts were unable to predict such course of inflation. In this paper, we offer a unique perspective on these events. First, we identify a few major influential factors that have altered significantly and reliably inflation since World War II. We then turn to looking at recent events in the light of these factors to try and extrapolate a likely trend for inflation in the coming years. Despite the dire economic challenges of World War II, the economy recovered quickly, the financial imbalances rectified in only a few years and inflation was tamed. Can the same thing be achieved after the pandemic? Our analysis suggests that this is highly improbable. Deep-rooted inflationary forces are at work because of the distortions that the economic order of the last 40 years has created. These distortions, exacerbated by the dual crisis, will take long to repair. We are then looking at an unsettled economic and inflationary future. The wise course of action to avoid a chaotic future requires that the US authorities withdraw from hands-on policies and instead, pave the way for the agile private sector to take the lead and adapt the economy to the changing conditions.
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Or, Etti, David Galbraith, and Anne Fennell. Exploring mechanisms involved in grape bud dormancy: Large-scale analysis of expression reprogramming following controlled dormancy induction and dormancy release. United States Department of Agriculture, December 2002. http://dx.doi.org/10.32747/2002.7587232.bard.

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The timing of dormancy induction and release is very important to the economic production of table grape. Advances in manipulation of dormancy induction and dormancy release are dependent on the establishment of a comprehensive understanding of biological mechanisms involved in bud dormancy. To gain insight into these mechanisms we initiated the research that had two main objectives: A. Analyzing the expression profiles of large subsets of genes, following controlled dormancy induction and dormancy release, and assessing the role of known metabolic pathways, known regulatory genes and novel sequences involved in these processes B. Comparing expression profiles following the perception of various artificial as well as natural signals known to induce dormancy release, and searching for gene showing similar expression patterns, as candidates for further study of pathways having potential to play a central role in dormancy release. We first created targeted EST collections from V. vinifera and V. riparia mature buds. Clones were randomly selected from cDNA libraries prepared following controlled dormancy release and controlled dormancy induction and from respective controls. The entire collection (7920 vinifera and 1194 riparia clones) was sequenced and subjected to bioinformatics analysis, including clustering, annotations and GO classifications. PCR products from the entire collection were used for printing of cDNA microarrays. Bud tissue in general, and the dormant bud in particular, are under-represented within the grape EST database. Accordingly, 59% of the our vinifera EST collection, composed of 5516 unigenes, are not included within the current Vitis TIGR collection and about 22% of these transcripts bear no resemblance to any known plant transcript, corroborating the current need for our targeted EST collection and the bud specific cDNA array. Analysis of the V. riparia sequences yielded 814 unigenes, of which 140 are unique (keilin et al., manuscript, Appendix B). Results from computational expression profiling of the vinifera collection suggest that oxidative stress, calcium signaling, intracellular vesicle trafficking and anaerobic mode of carbohydrate metabolism play a role in the regulation and execution of grape-bud dormancy release. A comprehensive analysis confirmed the induction of transcription from several calcium–signaling related genes following HC treatment, and detected an inhibiting effect of calcium channel blocker and calcium chelator on HC-induced and chilling-induced bud break. It also detected the existence of HC-induced and calcium dependent protein phosphorylation activity. These data suggest, for the first time, that calcium signaling is involved in the mechanism of dormancy release (Pang et al., in preparation). We compared the effects of heat shock (HS) to those detected in buds following HC application and found that HS lead to earlier and higher bud break. We also demonstrated similar temporary reduction in catalase expression and temporary induction of ascorbate peroxidase, glutathione reductase, thioredoxin and glutathione S transferase expression following both treatments. These findings further support the assumption that temporary oxidative stress is part of the mechanism leading to bud break. The temporary induction of sucrose syntase, pyruvate decarboxylase and alcohol dehydrogenase indicate that temporary respiratory stress is developed and suggest that mitochondrial function may be of central importance for that mechanism. These finding, suggesting triggering of identical mechanisms by HS and HC, justified the comparison of expression profiles of HC and HS treated buds, as a tool for the identification of pathways with a central role in dormancy release (Halaly et al., in preparation). RNA samples from buds treated with HS, HC and water were hybridized with the cDNA arrays in an interconnected loop design. Differentially expressed genes from the were selected using R-language package from Bioconductor project called LIMMA and clones showing a significant change following both HS and HC treatments, compared to control, were selected for further analysis. A total of 1541 clones show significant induction, of which 37% have no hit or unknown function and the rest represent 661 genes with identified function. Similarly, out of 1452 clones showing significant reduction, only 53% of the clones have identified function and they represent 573 genes. The 661 induced genes are involved in 445 different molecular functions. About 90% of those functions were classified to 20 categories based on careful survey of the literature. Among other things, it appears that carbohydrate metabolism and mitochondrial function may be of central importance in the mechanism of dormancy release and studies in this direction are ongoing. Analysis of the reduced function is ongoing (Appendix A). A second set of hybridizations was carried out with RNA samples from buds exposed to short photoperiod, leading to induction of bud dormancy, and long photoperiod treatment, as control. Analysis indicated that 42 genes were significant difference between LD and SD and 11 of these were unique.
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