Induction of through-thickness compressive residual stress fields in thin Al2024-T351 plates by laser shock processing

Laser Shock Processing (LSP) has been demonstrated as an emerging technique for the induction of RS’s fields in subsurface layers of relatively thick specimens. However, the LSP treatment of relatively thin specimens brings, as an additional consequence, the possible bending in a process of laser shock forming. This effect poses a new class of problems regarding the attainment of specified RS’s depth profiles in the mentioned type of sheets, and, what can be more critical, an overall deformation of the treated component. The analysis of the problem of LSP treatment for induction of tentatively through-thickness RS’s fields for fatigue life enhancement in relatively thin sheets in a way compatible with reduced overall workpiece deformation due to spring-back self-equilibration is envisaged in this paper. The coupled theoretical-experimental predictive approach developed by the authors has been applied to the specification of LSP treatments for achievement of RS's fields tentatively able to retard crack propagation on normalized specimens. A convergence between numerical code results and experimental results coming from direct RS's measurement is presented as a first step for the treatment of the normalized specimens under optimized conditions and verification of the crack retardation properties virtually induced.

A proposal for pellet production from residual woody biomass in the island of Majorca (Spain)

The use of residual biomass for energy purposes is of great interest in isolated areas like Majorca for waste reduction, energy sufficiency and renewable energies development. In addition, densification processes lead to easy-to-automate solid biofuels which additionally have higher energy density. The present study aims at (i) the estimation of the potential of residual biomass from woody crops as well as from agri-food and wood industries in Majorca, and (ii) the analysis of the optimal location of potential pellet plants by means of a GIS approach (location-allocation analysis) and a cost evaluation of the pellets production chain. The residual biomass potential from woody crops in Majorca Island was estimated at 35,874 metric tons dry matter (t DM) per year, while the wood and agri-food industries produced annually 21,494 t DM and 2717 t DM, respectively. Thus, there would be enough resource available for the installation of 10 pellet plants of 6400 t·year−1 capacity. These plants were optimally located throughout the island of Mallorca with a maximum threshold distance of 28 km for biomass transport from the production points. Values found for the biomass cost at the pellet plant ranged between 57.1 €·t−1 and 63.4 €·t−1 for biomass transport distance of 10 and 28 km. The cost of pelleting amounted to 56.7 €·t−1; adding the concepts of business fee, pellet transport and profit margin (15%), the total cost of pelleting was estimated at 116.6 €·t−1. The present study provides a proposal for pellet production from residual woody biomass that would supply up to 2.8% of the primary energy consumed by the domestic and services sector in the Balearic Islands.

Diseño de un módulo de IFMIF para irradiación de envolturas regeneradoras basadas en metales líquidos para reactores de fusión

La fusión nuclear es, hoy en día, una alternativa energética a la que la comunidad internacional dedica mucho esfuerzo. El objetivo es el de generar entre diez y cincuenta veces más energía que la que consume mediante reacciones de fusión que se producirán en una mezcla de deuterio (D) y tritio (T) en forma de plasma a doscientos millones de grados centígrados. En los futuros reactores nucleares de fusión será necesario producir el tritio utilizado como combustible en el propio reactor termonuclear. Este hecho supone dar un paso más que las actuales máquinas experimentales dedicadas fundamentalmente al estudio de la física del plasma. Así pues, el tritio, en un reactor de fusión, se produce en sus envolturas regeneradoras cuya misión fundamental es la de blindaje neutrónico, producir y recuperar tritio (fuel para la reacción DT del plasma) y por último convertir la energía de los neutrones en calor. Existen diferentes conceptos de envolturas que pueden ser sólidas o líquidas. Las primeras se basan en cerámicas de litio (Li2O, Li4SiO4, Li2TiO3, Li2ZrO3) y multiplicadores neutrónicos de Be, necesarios para conseguir la cantidad adecuada de tritio. Los segundos se basan en el uso de metales líquidos o sales fundidas (Li, LiPb, FLIBE, FLINABE) con multiplicadores neutrónicos de Be o el propio Pb en el caso de LiPb. Los materiales estructurales pasan por aceros ferrítico-martensíticos de baja activación, aleaciones de vanadio o incluso SiCf/SiC. Cada uno de los diferentes conceptos de envoltura tendrá una problemática asociada que se estudiará en el reactor experimental ITER (del inglés, “International Thermonuclear Experimental Reactor”). Sin embargo, ITER no puede responder las cuestiones asociadas al daño de materiales y el efecto de la radiación neutrónica en las diferentes funciones de las envolturas regeneradoras. Como referencia, la primera pared de un reactor de fusión de 4000MW recibiría 30 dpa/año (valores para Fe-56) mientras que en ITER se conseguirían <10 dpa en toda su vida útil. Esta tesis se encuadra en el acuerdo bilateral entre Europa y Japón denominado “Broader Approach Agreement “(BA) (2007-2017) en el cual España juega un papel destacable. Estos proyectos, complementarios con ITER, son el acelerador para pruebas de materiales IFMIF (del inglés, “International Fusion Materials Irradiation Facility”) y el dispositivo de fusión JT-60SA. Así, los efectos de la irradiación de materiales en materiales candidatos para reactores de fusión se estudiarán en IFMIF. El objetivo de esta tesis es el diseño de un módulo de IFMIF para irradiación de envolturas regeneradoras basadas en metales líquidos para reactores de fusión. El módulo se llamará LBVM (del inglés, “Liquid Breeder Validation Module”). La propuesta surge de la necesidad de irradiar materiales funcionales para envolturas regeneradoras líquidas para reactores de fusión debido a que el diseño conceptual de IFMIF no contaba con esta utilidad. Con objeto de analizar la viabilidad de la presente propuesta, se han realizado cálculos neutrónicos para evaluar la idoneidad de llevar a cabo experimentos relacionados con envolturas líquidas en IFMIF. Así, se han considerado diferentes candidatos a materiales funcionales de envolturas regeneradoras: Fe (base de los materiales estructurales), SiC (material candidato para los FCI´s (del inglés, “Flow Channel Inserts”) en una envoltura regeneradora líquida, SiO2 (candidato para recubrimientos antipermeación), CaO (candidato para recubrimientos aislantes), Al2O3 (candidato para recubrimientos antipermeación y aislantes) y AlN (material candidato para recubrimientos aislantes). En cada uno de estos materiales se han calculado los parámetros de irradiación más significativos (dpa, H/dpa y He/dpa) en diferentes posiciones de IFMIF. Estos valores se han comparado con los esperados en la primera pared y en la zona regeneradora de tritio de un reactor de fusión. Para ello se ha elegido un reactor tipo HCLL (del inglés, “Helium Cooled Lithium Lead”) por tratarse de uno de los más prometedores. Además, los valores también se han comparado con los que se obtendrían en un reactor rápido de fisión puesto que la mayoría de las irradiaciones actuales se hacen en reactores de este tipo. Como conclusión al análisis de viabilidad, se puede decir que los materiales funcionales para mantos regeneradores líquidos podrían probarse en la zona de medio flujo de IFMIF donde se obtendrían ratios de H/dpa y He/dpa muy parecidos a los esperados en las zonas más irradiadas de un reactor de fusión. Además, con el objetivo de ajustar todavía más los valores, se propone el uso de un moderador de W (a considerar en algunas campañas de irradiación solamente debido a que su uso hace que los valores de dpa totales disminuyan). Los valores obtenidos para un reactor de fisión refuerzan la idea de la necesidad del LBVM, ya que los valores obtenidos de H/dpa y He/dpa son muy inferiores a los esperados en fusión y, por lo tanto, no representativos. Una vez demostrada la idoneidad de IFMIF para irradiar envolturas regeneradoras líquidas, y del estudio de la problemática asociada a las envolturas líquidas, también incluida en esta tesis, se proponen tres tipos de experimentos diferentes como base de diseño del LBVM. Éstos se orientan en las necesidades de un reactor tipo HCLL aunque a lo largo de la tesis se discute la aplicabilidad para otros reactores e incluso se proponen experimentos adicionales. Así, la capacidad experimental del módulo estaría centrada en el estudio del comportamiento de litio plomo, permeación de tritio, corrosión y compatibilidad de materiales. Para cada uno de los experimentos se propone un esquema experimental, se definen las condiciones necesarias en el módulo y la instrumentación requerida para controlar y diagnosticar las cápsulas experimentales. Para llevar a cabo los experimentos propuestos se propone el LBVM, ubicado en la zona de medio flujo de IFMIF, en su celda caliente, y con capacidad para 16 cápsulas experimentales. Cada cápsula (24-22 mm de diámetro y 80 mm de altura) contendrá la aleación eutéctica LiPb (hasta 50 mm de la altura de la cápsula) en contacto con diferentes muestras de materiales. Ésta irá soportada en el interior de tubos de acero por los que circulará un gas de purga (He), necesario para arrastrar el tritio generado en el eutéctico y permeado a través de las paredes de las cápsulas (continuamente, durante irradiación). Estos tubos, a su vez, se instalarán en una carcasa también de acero que proporcionará soporte y refrigeración tanto a los tubos como a sus cápsulas experimentales interiores. El módulo, en su conjunto, permitirá la extracción de las señales experimentales y el gas de purga. Así, a través de la estación de medida de tritio y el sistema de control, se obtendrán los datos experimentales para su análisis y extracción de conclusiones experimentales. Además del análisis de datos experimentales, algunas de estas señales tendrán una función de seguridad y por tanto jugarán un papel primordial en la operación del módulo. Para el correcto funcionamiento de las cápsulas y poder controlar su temperatura, cada cápsula se equipará con un calentador eléctrico y por tanto el módulo requerirá también ser conectado a la alimentación eléctrica. El diseño del módulo y su lógica de operación se describe en detalle en esta tesis. La justificación técnica de cada una de las partes que componen el módulo se ha realizado con soporte de cálculos de transporte de tritio, termohidráulicos y mecánicos. Una de las principales conclusiones de los cálculos de transporte de tritio es que es perfectamente viable medir el tritio permeado en las cápsulas mediante cámaras de ionización y contadores proporcionales comerciales, con sensibilidades en el orden de 10-9 Bq/m3. Los resultados son aplicables a todos los experimentos, incluso si son cápsulas a bajas temperaturas o si llevan recubrimientos antipermeación. Desde un punto de vista de seguridad, el conocimiento de la cantidad de tritio que está siendo transportada con el gas de purga puede ser usado para detectar de ciertos problemas que puedan estar sucediendo en el módulo como por ejemplo, la rotura de una cápsula. Además, es necesario conocer el balance de tritio de la instalación. Las pérdidas esperadas el refrigerante y la celda caliente de IFMIF se pueden considerar despreciables para condiciones normales de funcionamiento. Los cálculos termohidráulicos se han realizado con el objetivo de optimizar el diseño de las cápsulas experimentales y el LBVM de manera que se pueda cumplir el principal requisito del módulo que es llevar a cabo los experimentos a temperaturas comprendidas entre 300-550ºC. Para ello, se ha dimensionado la refrigeración necesaria del módulo y evaluado la geometría de las cápsulas, tubos experimentales y la zona experimental del contenedor. Como consecuencia de los análisis realizados, se han elegido cápsulas y tubos cilíndricos instalados en compartimentos cilíndricos debido a su buen comportamiento mecánico (las tensiones debidas a la presión de los fluidos se ven reducidas significativamente con una geometría cilíndrica en lugar de prismática) y térmico (uniformidad de temperatura en las paredes de los tubos y cápsulas). Se han obtenido campos de presión, temperatura y velocidad en diferentes zonas críticas del módulo concluyendo que la presente propuesta es factible. Cabe destacar que el uso de códigos fluidodinámicos (e.g. ANSYS-CFX, utilizado en esta tesis) para el diseño de cápsulas experimentales de IFMIF no es directo. La razón de ello es que los modelos de turbulencia tienden a subestimar la temperatura de pared en mini canales de helio sometidos a altos flujos de calor debido al cambio de las propiedades del fluido cerca de la pared. Los diferentes modelos de turbulencia presentes en dicho código han tenido que ser estudiados con detalle y validados con resultados experimentales. El modelo SST (del inglés, “Shear Stress Transport Model”) para turbulencia en transición ha sido identificado como adecuado para simular el comportamiento del helio de refrigeración y la temperatura en las paredes de las cápsulas experimentales. Con la geometría propuesta y los valores principales de refrigeración y purga definidos, se ha analizado el comportamiento mecánico de cada uno de los tubos experimentales que contendrá el módulo. Los resultados de tensiones obtenidos, han sido comparados con los valores máximos recomendados en códigos de diseño estructural como el SDC-IC (del inglés, “Structural Design Criteria for ITER Components”) para así evaluar el grado de protección contra el colapso plástico. La conclusión del estudio muestra que la propuesta es mecánicamente robusta. El LBVM implica el uso de metales líquidos y la generación de tritio además del riesgo asociado a la activación neutrónica. Por ello, se han estudiado los riesgos asociados al uso de metales líquidos y el tritio. Además, se ha incluido una evaluación preliminar de los riesgos radiológicos asociados a la activación de materiales y el calor residual en el módulo después de la irradiación así como un escenario de pérdida de refrigerante. Los riesgos asociados al módulo de naturaleza convencional están asociados al manejo de metales líquidos cuyas reacciones con aire o agua se asocian con emisión de aerosoles y probabilidad de fuego. De entre los riesgos nucleares destacan la generación de gases radiactivos como el tritio u otros radioisótopos volátiles como el Po-210. No se espera que el módulo suponga un impacto medioambiental asociado a posibles escapes. Sin embargo, es necesario un manejo adecuado tanto de las cápsulas experimentales como del módulo contenedor así como de las líneas de purga durante operación. Después de un día de después de la parada, tras un año de irradiación, tendremos una dosis de contacto de 7000 Sv/h en la zona experimental del contenedor, 2300 Sv/h en la cápsula y 25 Sv/h en el LiPb. El uso por lo tanto de manipulación remota está previsto para el manejo del módulo irradiado. Por último, en esta tesis se ha estudiado también las posibilidades existentes para la fabricación del módulo. De entre las técnicas propuestas, destacan la electroerosión, soldaduras por haz de electrones o por soldadura láser. Las bases para el diseño final del LBVM han sido pues establecidas en el marco de este trabajo y han sido incluidas en el diseño intermedio de IFMIF, que será desarrollado en el futuro, como parte del diseño final de la instalación IFMIF. ABSTRACT Nuclear fusion is, today, an alternative energy source to which the international community devotes a great effort. The goal is to generate 10 to 50 times more energy than the input power by means of fusion reactions that occur in deuterium (D) and tritium (T) plasma at two hundred million degrees Celsius. In the future commercial reactors it will be necessary to breed the tritium used as fuel in situ, by the reactor itself. This constitutes a step further from current experimental machines dedicated mainly to the study of the plasma physics. Therefore, tritium, in fusion reactors, will be produced in the so-called breeder blankets whose primary mission is to provide neutron shielding, produce and recover tritium and convert the neutron energy into heat. There are different concepts of breeding blankets that can be separated into two main categories: solids or liquids. The former are based on ceramics containing lithium as Li2O , Li4SiO4 , Li2TiO3 , Li2ZrO3 and Be, used as a neutron multiplier, required to achieve the required amount of tritium. The liquid concepts are based on molten salts or liquid metals as pure Li, LiPb, FLIBE or FLINABE. These blankets use, as neutron multipliers, Be or Pb (in the case of the concepts based on LiPb). Proposed structural materials comprise various options, always with low activation characteristics, as low activation ferritic-martensitic steels, vanadium alloys or even SiCf/SiC. Each concept of breeding blanket has specific challenges that will be studied in the experimental reactor ITER (International Thermonuclear Experimental Reactor). However, ITER cannot answer questions associated to material damage and the effect of neutron radiation in the different breeding blankets functions and performance. As a reference, the first wall of a fusion reactor of 4000 MW will receive about 30 dpa / year (values for Fe-56) , while values expected in ITER would be <10 dpa in its entire lifetime. Consequently, the irradiation effects on candidate materials for fusion reactors will be studied in IFMIF (International Fusion Material Irradiation Facility). This thesis fits in the framework of the bilateral agreement among Europe and Japan which is called “Broader Approach Agreement “(BA) (2007-2017) where Spain plays a key role. These projects, complementary to ITER, are mainly IFMIF and the fusion facility JT-60SA. The purpose of this thesis is the design of an irradiation module to test candidate materials for breeding blankets in IFMIF, the so-called Liquid Breeder Validation Module (LBVM). This proposal is born from the fact that this option was not considered in the conceptual design of the facility. As a first step, in order to study the feasibility of this proposal, neutronic calculations have been performed to estimate irradiation parameters in different materials foreseen for liquid breeding blankets. Various functional materials were considered: Fe (base of structural materials), SiC (candidate material for flow channel inserts, SiO2 (candidate for antipermeation coatings), CaO (candidate for insulating coatings), Al2O3 (candidate for antipermeation and insulating coatings) and AlN (candidate for insulation coating material). For each material, the most significant irradiation parameters have been calculated (dpa, H/dpa and He/dpa) in different positions of IFMIF. These values were compared to those expected in the first wall and breeding zone of a fusion reactor. For this exercise, a HCLL (Helium Cooled Lithium Lead) type was selected as it is one of the most promising options. In addition, estimated values were also compared with those obtained in a fast fission reactor since most of existing irradiations have been made in these installations. The main conclusion of this study is that the medium flux area of IFMIF offers a good irradiation environment to irradiate functional materials for liquid breeding blankets. The obtained ratios of H/dpa and He/dpa are very similar to those expected in the most irradiated areas of a fusion reactor. Moreover, with the aim of bringing the values further close, the use of a W moderator is proposed to be used only in some experimental campaigns (as obviously, the total amount of dpa decreases). The values of ratios obtained for a fission reactor, much lower than in a fusion reactor, reinforce the need of LBVM for IFMIF. Having demonstrated the suitability of IFMIF to irradiate functional materials for liquid breeding blankets, and an analysis of the main problems associated to each type of liquid breeding blanket, also presented in this thesis, three different experiments are proposed as basis for the design of the LBVM. These experiments are dedicated to the needs of a blanket HCLL type although the applicability of the module for other blankets is also discussed. Therefore, the experimental capability of the module is focused on the study of the behavior of the eutectic alloy LiPb, tritium permeation, corrosion and material compatibility. For each of the experiments proposed an experimental scheme is given explaining the different module conditions and defining the required instrumentation to control and monitor the experimental capsules. In order to carry out the proposed experiments, the LBVM is proposed, located in the medium flux area of the IFMIF hot cell, with capability of up to 16 experimental capsules. Each capsule (24-22 mm of diameter, 80 mm high) will contain the eutectic allow LiPb (up to 50 mm of capsule high) in contact with different material specimens. They will be supported inside rigs or steel pipes. Helium will be used as purge gas, to sweep the tritium generated in the eutectic and permeated through the capsule walls (continuously, during irradiation). These tubes, will be installed in a steel container providing support and cooling for the tubes and hence the inner experimental capsules. The experimental data will consist of on line monitoring signals and the analysis of purge gas by the tritium measurement station. In addition to the experimental signals, the module will produce signals having a safety function and therefore playing a major role in the operation of the module. For an adequate operation of the capsules and to control its temperature, each capsule will be equipped with an electrical heater so the module will to be connected to an electrical power supply. The technical justification behind the dimensioning of each of these parts forming the module is presented supported by tritium transport calculations, thermalhydraulic and structural analysis. One of the main conclusions of the tritium transport calculations is that the measure of the permeated tritium is perfectly achievable by commercial ionization chambers and proportional counters with sensitivity of 10-9 Bq/m3. The results are applicable to all experiments, even to low temperature capsules or to the ones using antipermeation coatings. From a safety point of view, the knowledge of the amount of tritium being swept by the purge gas is a clear indicator of certain problems that may be occurring in the module such a capsule rupture. In addition, the tritium balance in the installation should be known. Losses of purge gas permeated into the refrigerant and the hot cell itself through the container have been assessed concluding that they are negligible for normal operation. Thermal hydraulic calculations were performed in order to optimize the design of experimental capsules and LBVM to fulfill one of the main requirements of the module: to perform experiments at uniform temperatures between 300-550ºC. The necessary cooling of the module and the geometry of the capsules, rigs and testing area of the container were dimensioned. As a result of the analyses, cylindrical capsules and rigs in cylindrical compartments were selected because of their good mechanical behavior (stresses due to fluid pressure are reduced significantly with a cylindrical shape rather than prismatic) and thermal (temperature uniformity in the walls of the tubes and capsules). Fields of pressure, temperature and velocity in different critical areas of the module were obtained concluding that the proposal is feasible. It is important to mention that the use of fluid dynamic codes as ANSYS-CFX (used in this thesis) for designing experimental capsules for IFMIF is not direct. The reason for this is that, under strongly heated helium mini channels, turbulence models tend to underestimate the wall temperature because of the change of helium properties near the wall. Therefore, the different code turbulence models had to be studied in detail and validated against experimental results. ANSYS-CFX SST (Shear Stress Transport Model) for transitional turbulence model has been identified among many others as the suitable one for modeling the cooling helium and the temperature on the walls of experimental capsules. Once the geometry and the main purge and cooling parameters have been defined, the mechanical behavior of each experimental tube or rig including capsules is analyzed. Resulting stresses are compared with the maximum values recommended by applicable structural design codes such as the SDC- IC (Structural Design Criteria for ITER Components) in order to assess the degree of protection against plastic collapse. The conclusion shows that the proposal is mechanically robust. The LBVM involves the use of liquid metals, tritium and the risk associated with neutron activation. The risks related with the handling of liquid metals and tritium are studied in this thesis. In addition, the radiological risks associated with the activation of materials in the module and the residual heat after irradiation are evaluated, including a scenario of loss of coolant. Among the identified conventional risks associated with the module highlights the handling of liquid metals which reactions with water or air are accompanied by the emission of aerosols and fire probability. Regarding the nuclear risks, the generation of radioactive gases such as tritium or volatile radioisotopes such as Po-210 is the main hazard to be considered. An environmental impact associated to possible releases is not expected. Nevertheless, an appropriate handling of capsules, experimental tubes, and container including purge lines is required. After one day after shutdown and one year of irradiation, the experimental area of the module will present a contact dose rate of about 7000 Sv/h, 2300 Sv/h in the experimental capsules and 25 Sv/h in the LiPb. Therefore, the use of remote handling is envisaged for the irradiated module. Finally, the different possibilities for the module manufacturing have been studied. Among the proposed techniques highlights the electro discharge machining, brazing, electron beam welding or laser welding. The bases for the final design of the LBVM have been included in the framework of the this work and included in the intermediate design report of IFMIF which will be developed in future, as part of the IFMIF facility final design.

Residual delay maps unveil global patterns of atmospheric nonlinearity and produce improved local forecasts

We use residual-delay maps of observational field data for barometric pressure to demonstrate the structure of latitudinal gradients in nonlinearity in the atmosphere. Nonlinearity is weak and largely lacking in tropical and subtropical sites and increases rapidly into the temperate regions where the time series also appear to be much noisier. The degree of nonlinearity closely follows the meridional variation of midlatitude storm track frequency. We extract the specific functional form of this nonlinearity, a V shape in the lagged residuals that appears to be a basic feature of midlatitude synoptic weather systems associated with frontal passages. We present evidence that this form arises from the relative time scales of high-pressure versus low-pressure events. Finally, we show that this nonlinear feature is weaker in a well regarded numerical forecast model (European Centre for Medium-Range Forecasts) because small-scale temporal and spatial variation is smoothed out in the grided inputs. This is significant, in that it allows us to demonstrate how application of statistical corrections based on the residual-delay map may provide marked increases in local forecast accuracy, especially for severe weather systems.

Elimination of residual metastatic prostate cancer after surgery and adjunctive cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) blockade immunotherapy

Cancer relapse after surgery is a common occurrence, most frequently resulting from the outgrowth of minimal residual disease in the form of metastases. We examined the effectiveness of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) blockade as an adjunctive immunotherapy to reduce metastatic relapse after primary prostate tumor resection. For these studies, we developed a murine model in which overt metastatic outgrowth of TRAMP-C2 (C2) prostate cancer ensues after complete primary tumor resection. Metastatic relapse in this model occurs reliably and principally within the draining lymph nodes in close proximity to the primary tumor, arising from established metastases present at the time of surgery. Using this model, we demonstrate that adjunctive CTLA-4 blockade administered immediately after primary tumor resection reduces metastatic relapse from 97.4 to 44%. Consistent with this, lymph nodes obtained 2 weeks after treatment reveal marked destruction or complete elimination of C2 metastases in 60% of mice receiving adjunctive anti-CTLA-4 whereas 100% of control antibody-treated mice demonstrate progressive C2 lymph node replacement. Our study demonstrates the potential of adjunctive CTLA-4 blockade immunotherapy to reduce cancer relapse emanating from minimal residual metastatic disease and may have broader implications for improving the capability of immunotherapy by combining such forms of therapy with other cytoreductive measures including surgery.

Antigen-driven CD4+ T cell and HIV-1 dynamics: Residual viral replication under highly active antiretroviral therapy

Antigen-induced stimulation of the immune system can generate heterogeneity in CD4+ T cell division rates capable of explaining the temporal patterns seen in the decay of HIV-1 plasma RNA levels during highly active antiretroviral therapy. Posttreatment increases in peripheral CD4+ T cell counts are consistent with a mathematical model in which host cell redistribution between lymph nodes and peripheral blood is a function of viral burden. Model fits to patient data suggest that, although therapy reduces HIV replication below replacement levels, substantial residual replication continues. This residual replication has important consequences for long-term therapy and the evolution of drug resistance and represents a challenge for future treatment strategies.

Changes in brain cell shape create residual extracellular space volume and explain tortuosity behavior during osmotic challenge

Diffusion of molecules in brain extracellular space is constrained by two macroscopic parameters, tortuosity factor λ and volume fraction α. Recent studies in brain slices show that when osmolarity is reduced, λ increases while α decreases. In contrast, with increased osmolarity, α increases, but λ attains a plateau. Using homogenization theory and a variety of lattice models, we found that the plateau behavior of λ can be explained if the shape of brain cells changes nonuniformly during the shrinking or swelling induced by osmotic challenge. The nonuniform cellular shrinkage creates residual extracellular space that temporarily traps diffusing molecules, thus impeding the macroscopic diffusion. The paper also discusses the definition of tortuosity and its independence of the measurement frame of reference.

NMR investigation of ferricytochrome c unfolding: Detection of an equilibrium unfolding intermediate and residual structure in the denatured state

Horse ferricytochrome c (cyt c) undergoes exchange of one of its axial heme ligands (Met-80) for one or more non-native ligands under denaturing conditions. We have used 1H NMR spectroscopy to detect two conformations of paramagnetic cyt c with non-native heme ligation through a range of urea concentrations. One non-native form is an equilibrium unfolding intermediate observed under partially denaturing conditions and is attributed to replacement of Met-80 with one or more Lys side chains. The second non-native form, in which the native Met ligand is replaced by a His, is observed under strongly denaturing conditions. Thermodynamic analysis of these data indicates a relatively small ΔG (17 kJ/mol) for the transition from native to the Lys-ligated intermediate and a significantly larger ΔG (47 kJ/mol) for the transition from native to the His-ligated species. Although CD and fluorescence data indicate that the equilibrium unfolding of cyt c is a two-state process, these NMR results implicate an intermediate with His-Lys ligation.

Characterization of residual structure in the thermally denatured state of barnase by simulation and experiment: Description of the folding pathway

Residual structure in the denatured state of a protein may contain clues about the early events in folding. We have simulated by molecular dynamics the denatured state of barnase, which has been studied by NMR spectroscopy. An ensemble of 104 structures was generated after 2 ns of unfolding and following for a further 2 ns. The ensemble was heterogeneous, but there was nonrandom, residual structure with persistent interactions. Helical structure in the C-terminal portion of helix α1 (residues 13–17) and in helix α2 as well as a turn and nonnative hydrophobic clustering between β3 and β4 were observed, consistent with NMR data. In addition, there were tertiary contacts between residues in α1 and the C-terminal portion of the β-sheet. The simulated structures allow the rudimentary NMR data to be fleshed out. The consistency between simulation and experiment inspires confidence in the methods. A description of the folding pathway of barnase from the denatured to the native state can be constructed by combining the simulation with experimental data from φ value analysis and NMR.

The Cellular Level of PR500, a Protein Complex Related to the 19S Regulatory Particle of the Proteasome, Is Regulated in Response to Stresses in Plants

In Arabidopsis seedlings and cauliflower florets, Rpn6 (a proteasome non-ATPase regulatory subunit) was found in two distinct protein complexes of ∼800 and 500 kDa, respectively. The large complex likely represents the proteasome 19S regulator particle (RP) because it displays the expected subunit composition and all characteristics. The small complex, designated PR500, shares at least three subunits with the “lid” subcomplex of 19S RP and is loosely associated with an hsp70 protein. In Arabidopsis COP9 signalosome mutants, PR500 was specifically absent or reduced to an extent that correlates with the severity of the mutations. Furthermore, PR500 was also diminished in response to potential protein-misfolding stresses caused by the heat shock and canavanine treatment. Immunofluorescence studies suggest that PR500 has a distinct localization pattern and is enriched in specific nuclear foci. We propose that PR500 may be evolved in higher plants to cope with the frequently encountered environmental stresses.

Mutations Affecting Induction of Glycolytic and Fermentative Genes during Germination and Environmental Stresses in Arabidopsis1

Expression of the alcohol dehydrogenase gene (ADH) of Arabidopsis is known to be induced by environmental stresses and regulated developmentally. We used a negative-selection approach to isolate mutants that were defective in regulating the expression of the ADH gene during seed germination; we then characterized three recessive mutants, aar1–1, aar1–2, and aar2–1, which belong to two complementation groups. In addition to their defects during seed germination, mutations in the AAR1 and AAR2 genes also affected anoxic and hypoxic induction of ADH and other glycolytic genes in mature plants. The aar1 and aar2 mutants were also defective in responding to cold and osmotic stress. The two allelic mutants aar1–1and aar1–2 exhibited different phenotypes under cold and osmotic stresses. Based on our results we propose that these mutants are defective in a late step of the signaling pathways that lead to increased expression of the ADH gene and glycolytic genes.

Translocation of PKN from the cytosol to the nucleus induced by stresses.

Effects of environmental stresses on the subcellular localization of PKN were investigated in NIH 3T3, BALB/c 3T3, and Rat-1 cells. The immunofluorescence of PKN resided prominently in the cytoplasmic region in nonstressed cells. When these cells were treated at 42 degrees C, there was a time-dependent decrease of the immunofluorescence of PKN in the cytoplasmic region that correlated with an increase within the nucleus as observed by confocal microscope. After incubation at 37 degrees C following beat shock, the immunofluorescence of PKN returned to the perinuclear and cytoplasmic regions from the nucleus. The nuclear translocation of PKN by heat shock was supported by the biochemical subcellular fractionation and immunoblotting. The nuclear localization of PKN was also observed when the cells were exposed to other stresses such as sodium arsenite and serum starvation. These results raise the possibility that there is a pathway mediating stress signals from the cytosol to the nucleus through PKN.