Innovation in the pharmaceutical industry:a study of the effects of regulation on the U.K. pharmaceutical industry

A history of government drug regulation and the relationship between the pharmaceutical companies in the U.K. and the licensing authority is outlined. Phases of regulatory stringency are identified with the formation of the Committees on Safety of Drugs and Medicines viewed as watersheds. A study of the impact of government regulation on industrial R&D activities focuses on the effects on the rate and direction of new product innovation. A literature review examines the decline in new chemical entity innovation. Regulations are cited as a major but not singular cause of the decline. Previous research attempting to determine the causes of such a decline on an empirical basis is given and the methodological problems associated with such research are identified. The U.K. owned sector of the British pharmaceutical industry is selected for a study employing a bottom-up approach allowing disaggregation of data. A historical background to the industry is provided, with each company analysed or a case study basis. Variations between companies regarding the policies adopted for R&D are emphasised. The process of drug innovation is described in order to determine possible indicators of the rate and direction of inventive and innovative activity. All possible indicators are considered and their suitability assessed. R&D expenditure data for the period 1960-1983 is subsequently presented as an input indicator. Intermediate output indicators are treated in a similar way and patent data are identified as a readily-available and useful source. The advantages and disadvantages of using such data are considered. Using interview material, patenting policies for most of the U.K. companies are described providing a background for a patent-based study. Sources of patent data are examined with an emphasis on computerised systems. A number of searches using a variety of sources are presented. Patent family size is examined as a possible indicator of an invention's relative importance. The patenting activity of the companies over the period 1960-1983 is given and the variation between companies is noted. The relationship between patent data and other indicators used is analysed using statistical methods resulting in an apparent lack of correlation. An alternative approach taking into account variations in company policy and phases in research activity indicates a stronger relationship between patenting activity, R&D Expenditure and NCE output over the period. The relationship is not apparent at an aggregated company level. Some evidence is presented for a relationship between phases of regulatory stringency, inventive and innovative activity but the importance of other factors is emphasised.

Industrial new product planning:a study of the sources of new product ideas and the development of new product proposals in the UK computer industry

This thesis presents a study of the sources of new product ideas and the development of new product proposals in an organisation in the UK Computer Industry. The thesis extends the work of von Hippel by showing how the phenomenon which he describes as "the Customer Active Paradigm for new product idea generation" can be observed to operate in this Industry. Furthermore, this thesis contrasts his Customer Active Paradigm with the more usually encountered Manufacturer Active Paradigm. In a second area, the thesis draws a number of conclusions relating to methods of market research, confirming existing observations and demonstrating the suitability of flexible interview strategies in certain circumstances. The thesis goes on to demonstrate the importance of free information flow within the organisation, making it more likely that sought and unsought opportunities can be exploited. It is shown that formal information flows and documents are a necessary but not sufficient means of influencing the formation of the organisation's dominant ideas on new product areas. The findings also link the work of Tushman and Katz on the role of "Gatekeepers" with the work of von Hippel by showing that the role of gatekeeper is particularly appropriate and useful to an organisation changing from Customer Active to Manufacturer Active methods of idea generation. Finally, the thesis provides conclusions relating to the exploitation of specific new product opportunities facing the sponsoring organisation.

Toward an atomistic understanding of the immune synapse:large-scale molecular dynamics simulation of a membrane-embedded TCR-pMHC-CD4 complex

T-cell activation requires interaction of T-cell receptors (TCR) with peptide epitopes bound by major histocompatibility complex (MHC) proteins. This interaction occurs at a special cell-cell junction known as the immune or immunological synapse. Fluorescence microscopy has shown that the interplay among one agonist peptide-MHC (pMHC), one TCR and one CD4 provides the minimum complexity needed to trigger transient calcium signalling. We describe a computational approach to the study of the immune synapse. Using molecular dynamics simulation, we report here on a study of the smallest viable model, a TCR-pMHC-CD4 complex in a membrane environment. The computed structural and thermodynamic properties are in fair agreement with experiment. A number of biomolecules participate in the formation of the immunological synapse. Multi-scale molecular dynamics simulations may be the best opportunity we have to reach a full understanding of this remarkable supra-macromolecular event at a cell-cell junction.

Modification of the B2-type matrix of aluminide diffusion coatings on nickel-base superalloys-bulk aluminide analogues

Pack aluminide coating is a useful method for conferring oxidation resistance on nickel-base superalloys. Nominally, these coatings have a matrix composed of a Ni-Al based B2-type phase (commonly denoted as Β). However, following high-temperature exposure in oxidative envi-ronments, aluminum is depleted from the coating. Aluminum depletion in turn, leads to de-stabilization of the Β phase, resulting in the formation of a characteristic lathlike Β-derivative microstructure. This article presents a transmission electron microscopy study of the formation of the lathlike Β-derivative microstructure using bulk nickel aluminides as model alloys. In the bulk nickel aluminides, the lathlike microstructure has been found to correspond to two distinct components: L10-type martensite and a new Β derivative. The new Β derivative is characterized and the conditions associated with the presence of this feature are identified and compared with those leading to the formation of the L10 martensitic phase. © 1995 The Minerals, Metals & Material Society.

The braids of the virgin: Taino roots of the early cult of La Virgen de la Caridad del Cobre in Cuba

Since her discovery in Cuban waters in 1611, La Virgen de la Caridad del Cobre (The Virgin of Charity) has become the leading transnational religious symbol for Cubans. The oldest stories about the appearance of La Virgen in Cuba suggest the presence of Cuban Taino in the early years of her cult. Yet historians have minimized Taino influence when examining Cuba's sixteenth and seventeenth centuries, pivotal years in the cult's development. This thesis demonstrated the significant role of the Taino in the formation of the early cult of La Virgen de la Caridad del Cobre, by employing revisionist historiography to the years between 1492 and 1687, to better understand the demographics and religious culture of Eastern Cuba, where the cult originated. It also found specific contributions from Taino religious culture in the myths, beliefs, and material culture associated with the early cult of La Virgen. ^

Biomimetic modeling of the nitrogen-centered radical postulated to occur during the inhibition of ribonucleotide reductases by 2'-azido-2'-deoxynucleotides

Ribonucleotide reductases (RNR) are essential enzymes that catalyze the reduction of ribonucleotides to 2'-deoxyribonucleotides, which is a critical step that produces precursors for DNA replication and repair. The inactivation of RNR, logically, would discontinue producing the precursors of the DNA of viral or cancer cells, which then would consequently end the cycle of DNA replication. Among different compounds that were found to be inhibitors of RNR, 2'-azido-2'-deoxynucleotide diphosphates (N3NDPs) have been investigated in depth as potent inhibitors of RNR. Decades of investigation has suggested that the inactivation of RNR by N3NDPs is a result of the formation of a nitrogen-centered radical (N·) that is covalently attached to the nucleotide at C3' and cysteine molecule C225 [3'-C(R-S-N·-C-OH)]. Biomimetic simulation reactions for the generation of the nitrogen-centered radicals similar to the one observed during the inactivation of the RNR by azionuclotides was investigated. The study included several modes: (i) theoretical calculation that showed the feasibility of the ring closure reaction between thiyl radicals and azido group; (ii) synthesis of the model azido nucleosides with a linker attached to C3' or C5' having a thiol or vicinal dithiol functionality; (iii) generation of the thiyl radical under both physiological and radiolysis conditions whose role is important in the initiation on RNR cascades; and (iv) analysis of the nitrogen-centered radical species formed during interaction between the thiyl radical and azido group by electron paramagnetic resonance spectroscopy (EPR). Characterization of the aminyl radical species formed during one electron attachment to the azido group of 2'-azido-2'-deoxyuridine and its stereospecifically labelled 1'-, 2'-, 3'-, 4'- or 5,6-[2H 2]-analogues was also examined. This dissertation gave insight toward understanding the mechanism of the formation of the nitrogen-centered radical during the inactivation of RNRs by azidonucleotides as well as the mechanism of action of RNRs that might provide key information necessary for the development of the next generation of antiviral and anticancer drugs.

Content -area teachers' perceptions of the effect of the meta -training on their instructional practices

The purpose of this study was to explore the content-area teachers' perceptions of the effect that compliance with the teacher training for content-area teachers of Limited English Proficient (LEP) students mandated by the Consent Decree had on their instructional practices within the content-area classroom. In order to provide an overview of the factors which had an effect on the content-area teachers' mandated training, various areas were reviewed: history of legislative actions that led up to the Consent Decree; stipulations set forth in the Consent Decree; Miami-Dade County Public Schools District LEP Plan including stipulations for teacher training; research on teacher training for teachers of language minority students; and the process of change. This descriptive study specifically addresses teachers' perceptions of the effectiveness of the mandated teacher training as it relates to language minority students. ^ Content-area teachers who had completed the mandated teacher training were surveyed using self-administered anonymous questionnaires mailed to their school sites. Questions focused on the teachers' perceptions of: students' need of second language instructional strategies within the content-area classroom; teacher training requirements mandated by the Consent Decree; and changes in their instructional practices as a result of the training. ^ Based on the responses of the subjects, the results of this study indicate the overall success of the training implemented to comply with the stipulations set forth in the Consent Decree. In general, the results indicate that the teachers perceive that they are ultimately in agreement with the mandated training. The results also indicate that the teachers perceive a need for second language strategies when working with language minority students. These results can serve as starting point for further research not only into teachers' perceptions of the effectiveness of training for teachers of language minority students but also into the outcomes of this teacher training as it is reflected within the classroom. ^

Cardiac Neural Crest Cells Affect the Development of the Myocardium and Conduction System in the Mouse

Neural crest cells originate from the dorsal most region of the embryonic neural tube. These cells migrate into several embryonic locations and differentiate into a variety of cell types. Cardiac neural crest (CNC) cells are a set of neural crest progenitors that aid in the proper formation of the cardiac septum, which separates the pulmonary from the systemic circulation. We have used Splotch mice to investigate whether the murine CNC cells play a role during the development oft he myocardium and the conduction system. Splotch mice carry a mutation in the P AX3 transcription factor, and display a problem in CNC cell migration. A scanning-electron-microscopy analysis of Splotch mutant-embryonic-hearts reveals abnormalities in the interventricular septum. In addition, the right and left ventricular cavities appear dilated relative to a wild type heart. Hoechst nuclei staining of Splotch heart cryosections demonstrates a decreased number of cardiomyocytes and a corresponding thinner ventricular wall. The absence of Connexin 40 in the ventricles of Splotch mutants, suggests conduction system defects. These results support the evidence that CNC cell signaling plays a role in modulating the growth and development of murine cardiomyocytes and their differentiation into conductile cells.

An alkalic volcanic suite of the Labrador Trough, Labrador

The Nimish Subgroup igneous suite is a linear belt of volcanic and plutonic rocks in the Dyke Lake area of the southern Labrador Trough. The volcanics are interbedded with the sediments of the Wishart and Sokoman Formations of the Aphebian aged, Knob Lake Group. The sokoman Formation forms a time stratigraphic horizon that separates the lower Petitsikapau Lake Formation from the upper Astray Lake formation of the Nimish Subgroup. The occurrence of these volcanics within the Knob Lake Group is unique relative to Labrador Trough stratigraphy, as elsewhere the Knob Lake Group is a dominantly sedimentary succession and volcanics are restricted to the younger Doublet Group. Stratigraphic relationships between the Nimish Subgroup and the Sokoman formation indicate contemporaneous volcanic, clastic and chemical sedimentary activity. The internal stratigraphy of the Sokoman Formation exhibits a three-fold subdivision that is broadly correlatable with similar subdivisions in the Schefferville "main ore zone", 30 miles to the northwest. A detailed facies and paleogeographic model relating the volcanic activity to iron formation deposition in the Dyke Lake is presented. The rocks of the Dyke Lake area have been affected by lower greenschist facies metamorphism during the Hudsonian orogenic event, circa 1735 my. Geochemical evidence indicates that the igneous rocks of the Nimish Subgroup have been metasomatized with large degrees of mobility in Na₂O, K₂O, CaO, MgO, SiO₂, FeO and Fe₂O₃ suspected. The "immobile trace elements", Ti, Zr, Nb, Y and Ga imply that the Nimish lavas are a mildly alkaline suite that has an alkali basalt-trachyandesite-comendite differentiation scheme. The rare earth element, REE, geochemistry of the Nimish Subgroup is supportive of the alkaline nature of the volcanics and has been used to model the fractional crystallization petrogenesis involved in the two volcanic cycles. The geological, geochemical and geophysical evidence indicates that the Nimish Subgroup lavas are possibly a rift facies, alkaline suite related to the tensional tectonic regime that preceeded the extrusion of voluminous tholeiitic lavas of the Doublet Group.

Structural and Biochemical Dissection of the Trehalose Biosynthetic Complex in Pathogenic Fungi

Trehalose is a non-reducing disaccharide essential for pathogenic fungal survival and virulence. The biosynthesis of trehalose requires the trehalose-6-phosphate synthase, Tps1, and trehalose-6-phosphate phosphatase, Tps2. More importantly, the trehalose biosynthetic pathway is absent in mammals, conferring this pathway as an ideal target for antifungal drug design. However, lack of germane biochemical and structural information hinders antifungal drug design against these targets.

In this dissertation, macromolecular X-ray crystallography and biochemical assays were employed to understand the structures and functions of proteins involved in the trehalose biosynthetic pathway. I report here the first eukaryotic Tps1 structures from Candida albicans (C. albicans) and Aspergillus fumigatus (A. fumigatus) with substrates or substrate analogs. These structures reveal the key residues involved in substrate binding and catalysis. Subsequent enzymatic assays and cellular assays highlight the significance of these key Tps1 residues in enzyme function and fungal stress response. The Tps1 structure captured in its transition-state with a non-hydrolysable inhibitor demonstrates that Tps1 adopts an “internal return like” mechanism for catalysis. Furthermore, disruption of the trehalose biosynthetic complex formation through abolishing Tps1 dimerization reveals that complex formation has regulatory function in addition to trehalose production, providing additional targets for antifungal drug intervention.

I also present here the structure of the Tps2 N-terminal domain (Tps2NTD) from C. albicans, which may be involved in the proper formation of the trehalose biosynthetic complex. Deletion of the Tps2NTD results in a temperature sensitive phenotype. Further, I describe in this dissertation the structures of the Tps2 phosphatase domain (Tps2PD) from C. albicans, A. fumigatus and Cryptococcus neoformans (C. neoformans) in multiple conformational states. The structures of the C. albicans Tps2PD -BeF3-trehalose complex and C. neoformans Tps2PD(D24N)-T6P complex reveal extensive interactions between both glucose moieties of the trehalose involving all eight hydroxyl groups and multiple residues of both the cap and core domains of Tps2PD. These structures also reveal that steric hindrance is a key underlying factor for the exquisite substrate specificity of Tps2PD. In addition, the structures of Tps2PD in the open conformation provide direct visualization of the conformational changes of this domain that are effected by substrate binding and product release.

Last, I present the structure of the C. albicans trehalose synthase regulatory protein (Tps3) pseudo-phosphatase domain (Tps3PPD) structure. Tps3PPD adopts a haloacid dehydrogenase superfamily (HADSF) phosphatase fold with a core Rossmann-fold domain and a α/β fold cap domain. Despite lack of phosphatase activity, the cleft between the Tps3PPD core domain and cap domain presents a binding pocket for a yet uncharacterized ligand. Identification of this ligand could reveal the cellular function of Tps3 and any interconnection of the trehalose biosynthetic pathway with other cellular metabolic pathways.

Combined, these structures together with significant biochemical analyses advance our understanding of the proteins responsible for trehalose biosynthesis. These structures are ready to be exploited to rationally design or optimize inhibitors of the trehalose biosynthetic pathway enzymes. Hence, the work described in this thesis has laid the groundwork for the design of Tps1 and Tps2 specific inhibitors, which ultimately could lead to novel therapeutics to treat fungal infections.

Physical and Genetic Analysis of the CUP1 Tandem Array in the Yeast Saccharomyces cerevisiae

The genomes of many strains of baker’s yeast, Saccharomyces cerevisiae, contain multiple repeats of the copper-binding protein Cup1. Cup1 is a member of the metallothionein family, and is found in a tandem array on chromosome VIII. In this thesis, I describe studies that characterized these tandem arrays and their mechanism of formation across diverse strains of yeast. I show that CUP1 arrays are an illuminating model system for observing recombination in eukaryotes, and describe insights derived from these observations.

In our first study, we analyzed 101 natural isolates of S. cerevisiae in order to examine the diversity of CUP1-containing repeats across different strains. We identified five distinct classes of repeats that contain CUP1. We also showed that some strains have only a single copy of CUP1. By comparing the sequences of all the strains, we were able to elucidate the mechanism of formation of the CUP1 tandem arrays, which involved unequal non-homologous recombination events starting from a strain that had only a single CUP1 gene. Our observation of CUP1 repeat formation allows more general insights about the formation of tandem repeats from single-copy genes in eukaryotes, which is one of the most important mechanisms by which organisms evolve.

In our second study, we delved deeper into our mechanistic investigations by measuring the relative rates of inter-homolog and intra-/inter-sister chromatid recombination in CUP1 tandem arrays. We used a diploid strain that is heterozygous both for insertion of a selectable marker (URA3) inside the tandem array, and also for markers at either end of the array. The intra-/inter-sister chromatid recombination rate turned out to be more than ten-fold greater than the inter-homolog rate. Moreover, we found that loss of the proteins Rad51 and Rad52, which are required for most inter-homolog recombination, did not greatly reduce recombination in the CUP1 tandem repeats. Additionally, we investigated the effects of elevated copper levels on the rate of each type of recombination at the CUP1 locus. Both types of recombination are increased at high concentrations of copper (as is known to be the case for CUP1 transcription). Furthermore, the inter-homolog recombination rate at the CUP1 locus is higher than the average over the genome during mitosis, but is lower than the average during meiosis.

The research described in Chapter 2 is published in 2014.

Strength of the Iberian intraplate lithosphere: Cenozoic deformations and seismicity

Previous studies about the strength of the lithosphere in the Iberia centre fail to resolve the depth of earthquakes because of the rheological uncertainties. Therefore, new contributions are considered (the crustal structure from a density model) and several parameters (tectonic regime, mantle rheology, strain rate) are checked in this paper to properly examine the role of lithospheric strength in the intraplate seismicity and the Cenozoic evolution. The strength distribution with depth, the integrated strength, the effective elastic thickness and the seismogenic thickness have been calculated by a finite element modelling of the lithosphere across the Central System mountain range and the bordering Duero and Madrid sedimentary basins. Only a dry mantle under strike-slip/extension and a strain rate of 10-15 s-1, or under extension and 10-16 s-1, causes a strong lithosphere. The integrated strength and the elastic thickness are lower in the mountain chain than in the basins. These anisotropies have been maintained since the Cenozoic and determine the mountain uplift and the biharmonic folding of the Iberian lithosphere during the Alpine deformations. The seismogenic thickness bounds the seismic activity in the upper–middle crust, and the decreasing crustal strength from the Duero Basin towards the Madrid Basin is related to a parallel increase in Plio–Quaternary deformations and seismicity. However, elasto–plastic modelling shows that current African–Eurasian convergence is resolved elastically or ductilely, which accounts for the low seismicity recorded in this region.

Strontium isotopes and sedimentology of a marine Triassic succession (upper Ladinian) of the westernmost Tethys, Spain

Chemical Stratigraphy, or the study of the variation of chemical elements within sedimentary sequences, has gradually become an experienced tool in the research and correlation of global geologic events. In this paper 87Sr/ 86Sr ratios of the Triassic marine carbonates (Muschelkalk facies) of southeast Iberian Ranges, Iberian Peninsula, are presented and the representative Sr-isotopic curve constructed for the upper Ladinian interval. The studied stratigraphic succession is 102 meters thick, continuous, and well preserved. Previous paleontological data from macro and micro, ammonites, bivalves, foraminifera, conodonts and palynological assemblages, suggest a Fassanian-Longobardian age (Late Ladinian). Although diagenetic minerals are present in small amounts, the elemental data content of bulk carbonate samples, especially Sr contents, show a major variation that probably reflects palaeoenvironmental changes. The 87Sr/86Sr ratios curve shows a rise from 0.707649 near the base of the section to 0.707741 and then declines rapidly to 0.707624, with a final values rise up to 0.70787 in the upper part. The data up to meter 80 in the studied succession is broadly concurrent with 87Sr/86Sr ratios of sequences of similar age and complements these data. Moreover, the sequence stratigraphic framework and its key surfaces, which are difficult to be recognised just based in the facies analysis, are characterised by combining variations of the Ca, Mg, Mn, Sr and CaCO3 contents

Structure, transports and transformations of the water masses in the Atlantic Subpolar Gyre

We discuss the distributions and transports of the main water masses in the North Atlantic Subpolar Gyre (NASPG) for the mean of the period 2002–2010 (OVIDE sections 2002–2010 every other year), as well as the inter-annual variability of the water mass structure from 1997 (4x and METEOR sections) to 2010. The water mass structure of the NASPG, quantitatively assessed by means of an Optimum MultiParameter analysis (with 14 water masses), was combined with the velocity fields resulting from previous studies using inverse models to obtain the water mass volume transports. We also evaluate the relative contribution to the Atlantic Meridional Overturning Circulation (AMOC) of the main water masses characterizing the NASPG, identifying the water masses that contribute to the AMOC variability. The reduction of the magnitude of the upper limb of the AMOC between 1997 and the 2000s is associated with the reduction in the northward transport of the Central Waters. This reduction of the northward flow of the AMOC is partially compensated by the reduction of the southward flow of the lower limb of the AMOC, associated with the decrease in the transports of Polar Intermediate Water and Subpolar Mode Water (SPMW) in the Irminger Basin. We also decompose the flow over the Reykjanes Ridge from the East North Atlantic Basin to the Irminger Basin (9.4 ± 4.7 Sv) into the contributions of the Central Waters (2.1 ± 1.8 Sv), Labrador Sea Water (LSW, 2.4 ± 2.0 Sv), Subarctic Intermediate Water (SAIW, 4.0 ± 0.5 Sv) and Iceland–Scotland Overflow Water (ISOW, 0.9 ± 0.9 Sv). Once LSW and ISOW cross over the Reykjanes Ridge, favoured by the strong mixing around it, they leave the Irminger Basin through the deep-to-bottom levels. The results also give insights into the water mass transformations within the NASPG, such as the contribution of the Central Waters and SAIW to the formation of the different varieties of SPMW due to air–sea interaction.

Tectono-metamorphic evolution of a nappe stack: A case study of the Swiss Alps

Fold-and-thrust belts are prominent structures that occur at the front of compressional orogens. To unravel the tectonic and metamorphic evolution of such complexes, kinematic investigations, quantitative microstructural analysis and geothermometry (calcite–graphite, calcite–dolomite) were performed on carbonate mylonites from thrust faults of the Helvetic nappe stack in Central Switzerland. Paleo-isotherms of peak temperature conditions and cooling stages (fission track) of the nappe pile were reconstructed in a vertical section and linked with the microstructural and kinematic evolution. Mylonitic microstructures suggest that under metamorphic conditions close to peak temperature, strain was highly localized within thrust faults where deformation temperatures spatially continuously increased in both directions, from N to S within each nappe and from top–down in the nappe stack, covering a temperature range of 180–380 °C. Due to the higher metamorphic conditions, thrusting of the lowermost nappe, the Doldenhorn nappe, was accompanied by a much more pronounced nappe internal ductile deformation of carbonaceous rock types than was the case for the overlying Wildhorn- and Gellihorn nappes. Ongoing thrusting brought the Doldenhorn nappe closer to the surface. The associated cooling resulted in a freezing in of the paleo-isotherms of peak metamorphic conditions. Contemporaneous shearing localized in the basal thrust, initially still in the ductile deformation regime and finally as brittle faulting and cataclasis inducing ultimately an inverse metamorphic zonation. With ongoing exhumation and the formation of the Helvetic antiformal nappe stack, a bending of large-scale tectonic structures (thrusts, folds), peak temperature isotherms and cooling isotherms occurred. While this local bending can directly be attributed to active deformation underneath the section investigated up to times of 2–3 ma, a more homogeneous uplift of the entire region is suggested for the very late and still active exhumation stage.