Proteostatic modulation in brain aging without associated Alzheimer's disease-and age-related neuropathological changes

(Phospho)proteomics of old-aged subjects without cognitive or behavioral symptoms, and without AD-neuropathological changes and lacking any other neurodegenerative alteration will increase understanding about the physiological state of human brain aging without associate neurological deficits and neuropathological lesions.(Phospho)proteomics using conventional label-free- and SWATH-MS (Sequential window acquisition of all theoretical fragment ion spectra mass spectrometry) has been assessed in the frontal cortex (FC) of individuals without NFTs, senile plaques (SPs) and age-related co-morbidities classified by age (years) in four groups; group 1 (young, 30-44); group 2 (middle-aged: MA, 45-52); group 3 (early-elderly, 64-70); and group 4 (late-elderly, 75-85).Protein levels and deregulated protein phosphorylation linked to similar biological terms/functions, but involving different individual proteins, are found in FC with age. The modified expression occurs in cytoskeleton proteins, membranes, synapses, vesicles, myelin, membrane transport and ion channels, DNA and RNA metabolism, ubiquitin-proteasome-system (UPS), kinases and phosphatases, fatty acid metabolism, and mitochondria. Dysregulated phosphoproteins are associated with the cytoskeleton, including microfilaments, actin-binding proteins, intermediate filaments of neurons and glial cells, and microtubules; membrane proteins, synapses, and dense core vesicles; kinases and phosphatases; proteins linked to DNA and RNA; members of the UPS; GTPase regulation; inflammation; and lipid metabolism. Noteworthy, protein levels of large clusters of hierarchically-related protein expression levels are stable until 70. However, protein levels of components of cell membranes, vesicles and synapses, RNA modulation, and cellular structures (including tau and tubulin filaments) are markedly altered from the age of 75. Similarly, marked modifications occur in the larger phosphoprotein clusters involving cytoskeleton and neuronal structures, membrane stabilization, and kinase regulation in the late elderly.Present findings may increase understanding of human brain proteostasis modifications in the elderly in the subpopulation of individuals not having AD neuropathological change and any other neurodegenerative change in any telencephalon region

Development of a State-Space Observer for Active Noise Control Systems

Active noise control (ANC) is a method of reducing the unwanted sound. This is realized by artificially generating canceling (secondary) source(s) of sound through detecting the unwanted (primary) noise and processing it by an electronic controller, so that when the secondary wave is superimposed on the primary wave the two destructively interfere and cancellation occurs at the observation point. ANC system is an active research area for its high demand especially in the acoustic noise and vibration control systems. A lot of work in modeling an ANC system involves the transfer function approach, but unfortunately this method allows observation at a single point or mode. It is of interest to measure the level of cancellation not only at the observer but also around it. Therefore, a state space approach would allow observation at multi modes simultaneously and became the subject of this research. This thesis is concerned with the study and development of a state-space model (SSM) for ANC system in on dimensional free-field medium instead of Finite Impulse Response (FIR) Models. In this work, the derivation of the SSM of each propagation path of ANC system is presented and hence the system is termed Feedforward state space control system with feedback inclusion single input single output (SISO) architecture. The criterions of success considered the evaluation process are the length of the propagation path, level of cancellation, convergence rate, number of modes of each path, and destructive interferences occur at the cancellation path. The secondary path of the ANC system is modeled by using the LMS algorithm to complete the design of the Filtered-X Least Mean Square (FXLMS) controller. Then the adaptive FXLMS controller is presented and incorporated with the proposed model for both Feedforward with / without the acoustic Feedback cases. As a result, the comparisons between the two cases are presented by mean of level of cancellation and convergence rate. The simulation results of the proposed model show that the level of the disturbance signal at ten modes along the primary path is decreasing as much as the modes go away from the source indicating that this model is suitable to build the mechanism of the ANC system which satisfies the relation between the wave dissipation against the number of modes which are distributed along the length of path. The derivation of the SSM gives the opportunity to extend the work furthermore to involve the derivation of a state-space optimal observer which is named State Space Least Mean Square (SSLMS) observer. This observer is employed to observe and monitor the pressure modes along the propagation path through simulating it in an offline structure i.e. without controller, or to observe the modes at the cancellation path through simulating the SSLMS in an on-line structure i.e. while the controller is converging. The comparison results between the real and observed modes of the secondary propagation show an accurate observing. Finally, the comparisons of the observed pressures of three modes along the cancellation path while the controller is converging (on-line structure) are shown with the mode which is located at the observer achieving the best cancellation

Improving continuous crystallisation using process analytical technologies: design of a novel periodic flow process

In this thesis novel configurations and operating strategies in the mixed suspension mixed product removal (MSMPR) crystalliser are investigated, aided by integrated process analytical technologies (PAT) and crystallisation informatics system (CryPRINS) tools. The MSMPR is an idealised crystalliser model that assumes: steady-state operation; well mixed suspension with no product classification, such that all volume elements contain a mixture of particles (small and large) and crystal size distribution (CSD) that is independent of location in the crystalliser and is identical of the product withdrawn; and uniform supersaturation thought, leading to constant nucleation and growth rates. Single-stage MSMPR designs with continuous recycle/recirculation and modified heat exchanger were investigated and found to minimise fouling, encrustation and transfer line blockages. In particular, a modified MSMPR with baffled heat exchanger was found to significantly reduce the temperature between incoming feed hot feed solution and the cooled crystalliser, leading to a significant reduction in fouling, encrustation and blockages. In addition, the concept of the periodic mixed suspension mixed product removal (PMSMPR) crystallisation process is demonstrated for the first time viz single- and multi-stage cascaded operations. This method of operation involves the periodic transfer of slurry (addition and withdrawal) at high flow rates from either a single stirred vessel or between a number of stirred vessels arranged in series. The PMSMPR is therefore characterised by periodic withdrawals of product slurry. Similar to the MSMPR, the product withdrawn from a PMSMPR has exactly the same composition as the vessel at the time of removal. The rapid withdrawal of slurry at high flow rates in PMSMPR operation leads to the prevention of particle sedimentation and blockage of transfer lines. The transfer of slurry (to/from) the PMSMPR is followed by a holding (or pause) period when no addition or withdrawal of slurry takes place. The holding period extends the mean residence time of the PMSMPR relative to a typical MSMPR, thereby increasing the yield and productivity of crystallisation as more time is allowed for consumption of available supersaturation viz crystal growth and nucleation. A state of controlled operation (SCO) in the periodic flow process, defined as a state of the system that maintains itself despite regular, but controlled disruptions was characterised using the PAT tools and CryPRINS within an intelligent decision support (IDS) framework. The crystallisation of paracetamol (PCM) from isopropyl alcohol (IPA) using different configurations of a single-stage continuous MSMPR crystalliser that incorporated continuous recycle and recirculation loop, and a novel design with baffled heat exchanger was investigated. Crystallisations of PCM-IPA carried out in the MSMPR without heat exchanger suffered from severe fouling, encrustation and blockage problems due to the high level of supersaturation (S = 1.39) in the crystalliser, which was required for the initial burst of nucleation to generate enough particles for later growth, as well as the large temperature difference between the incoming feed (45 oC) and the crystalliser (10 oC). Using the modified MSMPR design with baffled heat exchanger, the challenges of fouling, encrustation and blockage were significantly reduced due to the rapid lowering of the feed stream temperature prior to entering the crystalliser. In addition, the closed loop system led to conservation of material, which is a great benefit since large amounts of materials would otherwise be required if the MSMPR was operated with continuous product removal. This design is great for research purposes, in particular, to investigate process design and optimisation. Continuous crystallisation of PCM in the presence of hydroxyl propyl methyl cellulose (HPMC) additive was investigated in the modified MSMPR design with heat exchanger. HPMC was found to improve the crystallisation performance, leading to complete avoidance of fouling, encrustation and blockages at a concentration of 0.05 wt%. However, the yield of crystallisation was significantly reduced (28.0 %) compared to a control experiment (98.8 %, biased due to fouling/encrustation) performed without additive addition. Regardless, the productivity of crystallisation was more than four times that achieved in batch linear cooling (LC) (0.62 0.86 g/L-min) and batch automated dynamic nucleation control (ADNC) (0.24 0.25 g/L-min) runs. Aspects of the periodic flow crystallisation of single- and multi-component (co-crystals) molecular systems have also been examined to demonstrate the concept of state of controlled operation . The single component systems studied were PCM and glycine (GLY), each representative of compounds with slow and fast growth kinetics, respectively. The co-crystal systems investigated were urea-barbituric acid (UBA) and p Toluenesulfonamide-Triphenylphosphine oxide (p-TSA-TPPO). UBA is a polymorphic co-crystal system with three known forms (I, II and III). Form I UBA was successfully isolated in a three-stage periodic flow PMSMPR crystalliser. This study demonstrates the capability of periodic flow crystallisation for isolation of a desired polymorph from a mixture. p-TSA-TPPO exists in two known stoichiometric co-crystal forms, 1:1 and 3:2 mole ratio p-TSA-TPPO, respectively. The two crystalline forms exhibit solution mediated transformation, which proves to be a difficulty for separation. For this study, the implementation of temperature cycles in batch and flow control in semi-batch and periodic PMSMPR crystallisers were investigated to isolate pure 1:1 and 3:2 p-TSA-TPPO, respectively. Different regions of the ternary diagram of p-TSA, TPPO and acetonitrile (MeCN) were investigated. The desired co-crystal form was isolated all crystallisation platforms investigated. However, greater consistency was observed in the semi-batch and PMSMPR operations respectively. Periodic flow crystallisation in PMSMPR is a promising alternative to conventional continuous MSMPR operation, affording greater degrees of freedom operation, slightly narrower RTD profiles, consistent product crystal quality (size, shape and distribution), longer mean residence times, higher yield and productivity and significant reduction in fouling, encrustation and transfer line blockages over prolonged operating periods. Furthermore, the PMSMPR is a versatile platform that can be used to investigate a range of different molecular systems. Relative to batch operation, the PMSMPR can operate close to equilibrium, however, this is dependent on the system kinetics. In addition, retrofitting of batch crystallisers to operate as PMSMPRS fairly simple and require only subtle changes to the existing design space. The integrated array of PAT sensors consisted of attenuated total reflectance ultra violet/visible spectroscopy (ATR-UV/vis), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), focused beam reflectance measurement (FBRM), particle vision microscopy (PVM) and Raman spectroscopy. The results from the studies reported here illustrate very well the use of PAT and information system tools together to determine when the continuous and periodic MSMPR operations reaches a steady-state or state of controlled operation (i.e. periodic steady-state). These tools provided a better understanding of the variables and operating procedures that influence the two types of operations

A framework for understanding and controlling batch cooling crystallization

In taking a different view of crystallization dynamics, this thesis reveals a new framework for addressing a prevalent process engineering challenge: control over the size of crystals produced by batch cooling crystallization. The thesis divides roughly into halves. In the first half, the crystal size control problem is introduced and the proposed framework for addressing this problem—termed the mass-count (MC) framework—is developed. This new framework is laid out along side the population balance (PB) framework, which is the prevailing framework for modeling crystallization dynamics and addressing the crystal size control problem. In putting the proposed and established frameworks side by side, the intent is not to say that one or the other is correct. Rather, the point is to show that they are different perspectives that facilitate different control approaches. The PB framework is built up from first principles; it is intellectually stimulating and mathematically complete, but it has a drawback for application: it does not directly enable feedback control. The MC framework, on the other hand, takes a less detailed view of crystallization dynamics and does not connect to crystallization theory as directly; it is also more conducive to application. In the second half of the thesis, the utility of the MC framework is put to the test. The framework is first applied to understand and model the crystallization dynamics for two widely different systems: darapskite salt crystallization from water and paracetamol crystallization from ethanol. Once the dynamics have been modeled, the framework is then used to develop feedback control schemes. These schemes are applied to both experimental systems and, in both cases, crystal size control is demonstrated.Ph.D

Uso del probiótico Shewanella putrefaciens Pdp11 en el cultivo de Solea senegalensis: implicaciones sobre la microbiota intestinal

Actualmente, la acuicultura es una industria en expansión. Entre las especies que se están introduciendo destaca el lenguado, Solea senegalensis, cuyo cultivo en cautividad no acaba de ser económicamente rentable debido fundamentalmente a las mortalidades producidas por infecciones bacterianas. Los antibióticos presentan una serie de limitaciones ya conocidas, siendo los probióticos una de las opciones más consideradas. Estudios previos en nuestro laboratorio han mostrado que la cepas S. putrefaciens Pdp11 (Pdp11) es capaz de inhibir el crecimiento de bacterias patógenas de peces, por lo que se planteó su uso como probiótico y el estudio de los efectos sobre la fisiología de S. senegalensis así como la potencialidad en la prevención de infecciones. La incorporación de Pdp11 a la dieta de S. senegalensis induce una modulación de la microbiota intestinal y siendo capaz de colonizar el intestino del animal y presentando mayor diversidad en la microbiota, menor susceptibilidad a infecciones y mejor composición corporal, sobre todo relativo a la cantidad de ácidos grasos beneficiosos. El estudio de la forma más adecuada de administración del probiótico mostró que las células no liofilizadas tienen un tiempo de permanencia superior en el intestino y su administración aumenta la diversidad de la microbiota intestinal al tiempo que reduce la presencia de patógenos oportunistas en la misma. Por este motivo, se consideró la forma no liofilizada como la más idónea para los estudios posteriores. La administración de Pdp11 a ejemplares de S. senegalensis sometidos a estrés por alta densidad o tratamiento con oxitetraciclina (OTC) permitió determinar la presencia de una microbiota intestinal y expresión génica diferencial en dichos ejemplares, con sobreexpresión de genes relacionados con el sistema inmune, mejoría de la histología intestinal y protección frente a infecciones por V. harveyi y V. parahaemolyticus. Por otro lado, la administración de OTC produce cambios radicales en la microbiota, pero éstos son menos acusados cuando va junto con Pdp11 apareciendo en la microbiota especies beneficiosas y desapareciendo otras patógenas. Además, el aumento de la expresión de genes relacionados con el estrés oxidativo observado tras la administración de OTC no ocurre cuando se aplica simultáneamente con Pdp11. Por último, se ha estudiado el efecto de la administración del probiótico durante la fase larvaria, fase muy susceptible a infecciones. La aplicación de forma continuada durante esta fase supone una modulación de la microbiota la cual se enriquece en especies de Vibrio. Del mismo modo, la aplicación sólo durante la fase de metamorfosis (desde el día 10 al 30 después de la eclosión) produce una modulación de la microbiota desde el inicio y a la ausencia de patógenos como P. damselae subsp piscicida. Por tanto, Pdp11 se puede considerar como un probiótico adecuado para S. senegalensis, que limita la presencia de patógenos en el intestino, atenúa los efectos del estrés y protege en estadios larvarios

Expresión de Aromatasa Cerebral y Gonadal y cambios histológicos gonadales como biomarcadores de exposición a contaminantes ambientales.

Tesis (Grado Doctor en Ciencias Biológicas)--Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Lugar de Trabajo: Cátedra de Diversidad Animal II. Facultad de Ciencias Exactas, Físicas y Naturales. Universidad Nacional de Córdoba. 2013- 151 h. con Apéndices + CD. tabls.; ils.; grafs. Contiene Referencia Bibliográfica, y Copia de Publicaciones Derivadas de la Tesis. Abstract en español e inglés.El objetivo general de la presente Tesis Doctoral fue realizar un análisis integral, que contemple el efecto de la exposición a contaminantes ambientales sobre biomarcadores de diferentes niveles de organización (moleculares, histológicos y somáticos) en peces. Para ello se evaluó la respuesta de los genes cyp19a1 (aromatasa), la histología de las gónadas y los índices somáticos en machos de la especie íctica autóctona Jenynsia multidentata tanto en condiciones de laboratorio como a campo. En bioensayos se analizaron los cambios en los biomarcadores frente a la exposición a 17β‐estradiol, 17α‐etinilestradiol, 4n‐nonilfenol y 17α‐metiltestosterona; mientras que a campo se realizó una caracterización de la calidad del agua y de los niveles de expresión de los genes cyp19a1, y se compararon las respuestas de los biomarcadores entre dos sitios con diferente calidad de agua de la cuenca del Río Suquía. Los resultados obtenidos en esta tesis demuestran que la aromatasa cerebral responde como un biomarcador sensible frente a la exposición a compuestos que presentan actividad estrogénica en condiciones de laboratorio, respondiendo incluso ante la concentración más baja de estos compuestos.El análisis histológico de los testículos reveló un gran impacto de la exposición a los diferentes químicos sobre este órgano. A diferencia de lo observado a nivel molecular y tisular, los índices somáticos (Factor de Condición, Índice Hepatosomático e Índice Gonadosomático) no mostraron marcados cambios frente a la exposición a compuestos estrogénicos. A campo se observaron fluctuaciones de la expresión de la aromatasa cerebral a lo largo del año, siendo máxima durante la estación reproductiva en ambos sitios de muestreo, sugiriendo su implicancia en el control del ciclo reproductivo. Se observó un desfasaje en el incremento de la expresión del gen entre los sitios, registrándose el aumento con un mes de retraso en el sitio contaminado respecto al de referencia,lo cual podría estar relacionado con desfasajes en el comienzo del ciclo reproductivo. Los índices somáticos revelaron efectos adversos debidos a la exposición a poluentes ambientales y la histología de los testículos se vio severamente afectada en el sitio contaminado,evidenciando lesiones que incluyeron la desorganización del tejido testicular, alteraciones severas de las células germinales, presencia de células vacuolares, desincronización del proceso de espermatogénesis y necrosis. Las concentraciones de los compuestos utilizados para los ensayos de laboratorio se encuentran dentro del rango detectado en aguas superficiales de Argentina,demostrando que los peces están expuestos a estos niveles de contaminantes en el ambiente. Los cambios deletéreos observados en J. multidentata frente a la exposición crónica a contaminantes ambientales (tanto en condiciones de laboratorio como a campo), podrían afectar su capacidad reproductiva y representar una amenaza para la especie

Fisiología de la reproducción del lenguado senegalés (Solea senegalensis): mecanismos endocrinos y aplicaciones en acuicultura.

RESUMEN En el presente trabajo investigamos algunos de los mecanismos endocrinos implicados en el crecimiento oocitario del lenguado senegalés (Solea senegalensis), así como el desarrollo de métodos para el control de su reproducción en cautividad. La tesis se dividió en tres partes diferentes: (i) Capítulo 1: Introducción General donde se resume el estado actual de conocimientos sobre los mecanismos endocrinos implicados en la reproducción de teleósteos, y se introduce al lector en los beneficios y problemas actuales de la acuicultura de peces planos, especialmente del lenguado senegalés (Solea senegalensis); (ii) Capítulos 2 y 3, donde se investiga el proceso de vitelogénesis en el lenguado senegalés. En el Capítulo 2, se caracteriza el ADN complementario de ambas isoformas del receptor de lipoproteínas de muy baja densidad/vitelogenina (Vtg), vtgr, y de una nueva proteína de unión a ácidos grasos (FABP), FABP11, encontrada exclusivamente en peces teleósteos. El análisis de expresión de estos genes en ovario de lenguado senegalés, sugiere que el nivel de los transcritos de vtgr puede ser utilizado como marcador molecular para determinar el número de oocitos reclutados en vitelogénesis, mientras que los niveles de expresión de fabp11 pueden funcionar como un marcador molecular muy útil para determinar los procesos celulares y factores ambientales que regulan el proceso de la atresia folicular. En el Capítulo 3, se valida un ELISA homologo para determinar la concentración de Vtg en plasma. Mediante este análisis se correlacionan las variaciones anuales de Vtg con los niveles de 17-estradiol, índice gonadosomático y porcentaje de oocitos en diferente estadio de desarrollo oocitario; y (iii) Capítulos 4 y 5, donde se investiga la aplicación de diferentes métodos hormonales para inducir a la ovulación y a la espermiación en lenguado senegalés en condiciones de cautividad. En el Capítulo 4, se tratan machos y hembras con un agonista de la hormona liberadora de gonadotropinas (GnRHa), tanto mediante inyecciones sucesivas como mediante implantes de liberación sostenida. El tratamiento con GnRHa resulta eficaz para inducir la ovulación y la puesta en hembras F1 incapaces de reproducirse en cautividad. En cambio, el mismo tratamiento con GnRHa en machos es aparentemente ineficaz para aumentar la producción de esperma y/o estimular la espermatogénesis. En el Capítulo 5, el tratamiento de machos adultos con GnRHa en combinación con 11-ketoandrostenediona (OA), precursor de 11-ketotestosterona (11-KT), acelera la espermatogénesis y aumenta los niveles plasmáticos de los metabolitos 5β-reducidos de 17α,20β-dihidroxi-4-pregnan-3-ona (17,20βP), esteroide implicado en la maduración del esperma en salmónidos. Además, la motilidad del esperma producido por los machos tratados con GnRHa+OA, se ve duplicada respecto al esperma producido por el grupo control o el grupo tratado sólo con GnRHa. Por tanto, el tratamiento de machos de lenguado senegalés con GnRHa+OA puede ser empleado como potencial terapia hormonal para mejorar disfunciones reproductivas de machos de lenguado senegalés en condiciones de cautividad. En resumen, la presente tesis ha contribuido a profundizar en el conocimiento de los mecanismos endocrinos implicados en la gametogénesis del lenguado senegalés, y ha establecido algunos protocolos para la inducción de la ovulación y la espermiogénesis a través de terapias hormonales en esta especie de elevado interés comercial. __________________________________________________________________________________________________The present work was aimed at the investigation of some endocrine mechanisms involved in oocyte growth in the Senegalese sole (Solea senegalensis), as well as to develop methods for the control of reproduction of this species in captivity. The thesis was divided into three separate parts: (i) Chapter 1, a General Introduction where it was reviewed the endocrine regulation of reproduction in teleosts, and it introduced the reader into the significance and current problems in the culture of flatfish, particularly the Senegalese sole; (ii) Chapters 2 and 3, in which it was investigated the vitellogenic process of Senegalese sole. In chapter 2, it is described the isolation of complementary DNAs encoding two isoforms of very low-density lipoprotein/vitellogenin receptor (VtgR) and a novel fatty acid-binding protein (FABP), FAB11. The analysis of expression of these genes suggested that the the level of vtgr transcripts in the ovary may be used as a functional marker to quantify the number of oocytes recruited for vitellogenesis, while that of fabp11 may be a very useful molecular marker for determining cellular events and environmental factors that regulate follicular atresia. In chapter 3, an homologous EIA to quantify plasma vitellogenin was developed, and annual plasma levels of vitellogenin were correlated with changes in plasma 17-estradiol, gonadosomatic index, and percentage of oocyte at different developmental stage in the ovary; and (iii) Chapters 4 and 5, in which it was investigated the induction of ovulation and spermiation in Senegalese sole. In chapter 4, males and females were treated with gonadotropin-releasing hormone agonist (GnRHa). Treatment was able to induce ovulation and spawning of F1 females that often fail to reproduce. However, this treatment was ineffective in males. Further studies were performed in chapter 5, where GnRHa treatment in combination with a biosynthetic precursor of 11-ketotestosterone, stimulated spermatogenesis, increasing the production of conjugated forms of 17,20-dihydroxy-4-pregnen-3-one, enhancing the motility of spermatozoa by 2-fold. In conclusion, the present work has contributed with novel information on the endocrine mechanisms involved in gametogenesis in Senegalese sole, and has established some protocols for the induction of ovulation and spermiation in this species through hormonal therapies

Iterative learning control of crystallisation systems

Under the increasing pressure of issues like reducing the time to market, managing lower production costs, and improving the flexibility of operation, batch process industries thrive towards the production of high value added commodity, i.e. specialty chemicals, pharmaceuticals, agricultural, and biotechnology enabled products. For better design, consistent operation and improved control of batch chemical processes one cannot ignore the sensing and computational blessings provided by modern sensors, computers, algorithms, and software. In addition, there is a growing demand for modelling and control tools based on process operating data. This study is focused on developing process operation data-based iterative learning control (ILC) strategies for batch processes, more specifically for batch crystallisation systems. In order to proceed, the research took a step backward to explore the existing control strategies, fundamentals, mechanisms, and various process analytical technology (PAT) tools used in batch crystallisation control. From the basics of the background study, an operating data-driven ILC approach was developed to improve the product quality from batch-to-batch. The concept of ILC is to exploit the repetitive nature of batch processes to automate recipe updating using process knowledge obtained from previous runs. The methodology stated here was based on the linear time varying (LTV) perturbation model in an ILC framework to provide a convergent batch-to-batch improvement of the process performance indicator. In an attempt to create uniqueness in the research, a novel hierarchical ILC (HILC) scheme was proposed for the systematic design of the supersaturation control (SSC) of a seeded batch cooling crystalliser. This model free control approach is implemented in a hierarchical structure by assigning data-driven supersaturation controller on the upper level and a simple temperature controller in the lower level. In order to familiarise with other data based control of crystallisation processes, the study rehearsed the existing direct nucleation control (DNC) approach. However, this part was more committed to perform a detailed strategic investigation of different possible structures of DNC and to compare the results with that of a first principle model based optimisation for the very first time. The DNC results in fact outperformed the model based optimisation approach and established an ultimate guideline to select the preferable DNC structure. Batch chemical processes are distributed as well as nonlinear in nature which need to be operated over a wide range of operating conditions and often near the boundary of the admissible region. As the linear lumped model predictive controllers (MPCs) often subject to severe performance limitations, there is a growing demand of simple data driven nonlinear control strategy to control batch crystallisers that will consider the spatio-temporal aspects. In this study, an operating data-driven polynomial chaos expansion (PCE) based nonlinear surrogate modelling and optimisation strategy was presented for batch crystallisation processes. Model validation and optimisation results confirmed this approach as a promise to nonlinear control. The evaluations of the proposed data based methodologies were carried out by simulation case studies, laboratory experiments and industrial pilot plant experiments. For all the simulation case studies a detailed mathematical models covering reaction kinetics and heat mass balances were developed for a batch cooling crystallisation system of Paracetamol in water. Based on these models, rigorous simulation programs were developed in MATLAB®, which was then treated as the real batch cooling crystallisation system. The laboratory experimental works were carried out using a lab scale system of Paracetamol and iso-Propyl alcohol (IPA). All the experimental works including the qualitative and quantitative monitoring of the crystallisation experiments and products demonstrated an inclusive application of various in situ process analytical technology (PAT) tools, such as focused beam reflectance measurement (FBRM), UV/Vis spectroscopy and particle vision measurement (PVM) as well. The industrial pilot scale study was carried out in GlaxoSmithKline Bangladesh Limited, Bangladesh, and the system of experiments was Paracetamol and other powdered excipients used to make paracetamol tablets. The methodologies presented in this thesis provide a comprehensive framework for data-based dynamic optimisation and control of crystallisation processes. All the simulation and experimental evaluations of the proposed approaches emphasised the potential of the data-driven techniques to provide considerable advances in the current state-of-the-art in crystallisation control

La familia génica FTF determina el patrón de colonización y la virulencia en Fusarium oxysporum

[ES] La familia génica FTF (Fusarium Transcription Factor) comprende un gen de copia única, FTF2, que está presente en todos los ascomicetos filamentosos analizados, y varias copias de FTF1, que es exclusivo de Fusarium oxysporum. Un sistema de silenciamiento de genes mediado por ARN ha sido desarrollado para atenuar específicamente los ARNm producidos por todos los genes FTF, probándose en dos formae speciales: F. oxysporum f. sp. phaseoli (cuyo hospedador es la judía) y F. oxysporum f. sp. lycopersici (cuyo hospedador es el tomate). La disminución de la expresión de FTF da lugar a una marcada reducción en la virulencia, una expresión reducida de varios genes SIX (Secreted In Xylem), la familia de efectores mejor estudiada en F. oxysporum, y menores niveles de ARNm de SGE1 (SIX Gene Expression 1), el presunto regulador de la expresión de algunos genes SIX. Por otra parte, los mutantes silenciados muestran un patrón de colonización de la planta huésped similar al mostrado por las estirpes carentes de copias de FTF1 (estirpes poco virulentas), caracterizado por una colonización de los haces xilemáticos lenta y de baja eficiencia, y una fuerte y temprana inducción de la resistencia sistémica adquirida (SAR) en el huésped