Advances in global instability computations: from incompressible to hypersonic flow

Esta tesis constituye un gran avance en el conocimiento del estudio y análisis de inestabilidades hidrodinámicas desde un punto de vista físico y teórico, como consecuencia de haber desarrollado innovadoras técnicas para la resolución computacional eficiente y precisa de la parte principal del espectro correspondiente a los problemas de autovalores (EVP) multidimensionales que gobiernan la inestabilidad de flujos con dos o tres direcciones espaciales inhomogéneas, denominados problemas de estabilidad global lineal. En el contexto del trabajo de desarrollo de herramientas computacionales presentado en la tesis, la discretización mediante métodos de diferencias finitas estables de alto orden de los EVP bidimensionales y tridimensionales que se derivan de las ecuaciones de Navier-Stokes linealizadas sobre flujos con dos o tres direcciones espaciales inhomogéneas, ha permitido una aceleración de cuatro órdenes de magnitud en su resolución. Esta mejora de eficiencia numérica se ha conseguido gracias al hecho de que usando estos esquemas de diferencias finitas, técnicas eficientes de resolución de problemas lineales son utilizables, explotando el alto nivel de dispersión o alto número de elementos nulos en las matrices involucradas en los problemas tratados. Como más notable consecuencia cabe destacar que la resolución de EVPs multidimensionales de inestabilidad global, que hasta la fecha necesitaban de superordenadores, se ha podido realizar en ordenadores de sobremesa. Además de la solución de problemas de estabilidad global lineal, el mencionado desarrollo numérico facilitó la extensión de las ecuaciones de estabilidad parabolizadas (PSE) lineales y no lineales para analizar la inestabilidad de flujos que dependen fuertemente en dos direcciones espaciales y suavemente en la tercera con las ecuaciones de estabilidad parabolizadas tridimensionales (PSE-3D). Precisamente la capacidad de extensión del novedoso algoritmo PSE-3D para el estudio de interacciones no lineales de los modos de estabilidad, desarrollado íntegramente en esta tesis, permite la predicción de transición en flujos complejos de gran interés industrial y por lo tanto extiende el concepto clásico de PSE, el cuál ha sido empleado exitosamente durante las pasadas tres décadas en el mismo contexto para problemas de capa límite bidimensional. Típicos ejemplos de flujos incompresibles se han analizado en este trabajo sin la necesidad de recurrir a restrictivas presuposiciones usadas en el pasado. Se han estudiado problemas vorticales como es el caso de un vórtice aislado o sistemas de vórtices simulando la estela de alas, en los que la homogeneidad axial no se impone y así se puede considerar la difusión viscosa del flujo. Además, se ha estudiado el chorro giratorio turbulento, cuya inestabilidad se utiliza para mejorar las características de funcionamiento de combustores. En la tesis se abarcan adicionalmente problemas de flujos compresibles. Se presenta el estudio de inestabilidad de flujos de borde de ataque a diferentes velocidades de vuelo. También se analiza la estela formada por un elemento rugoso aislado en capa límite supersónica e hipersónica, mostrando excelentes comparaciones con resultados obtenidos mediante simulación numérica directa. Finalmente, nuevas inestabilidades se han identificado en el flujo hipersónico a Mach 7 alrededor de un cono elíptico que modela el vehículo de pruebas en vuelo HIFiRE-5. Los resultados comparan favorablemente con experimentos en vuelo, lo que subraya aún más el potencial de las metodologías de análisis de estabilidad desarrolladas en esta tesis. ABSTRACT The present thesis constitutes a step forward in advancing the frontiers of knowledge of fluid flow instability from a physical point of view, as a consequence of having been successful in developing groundbreaking methodologies for the efficient and accurate computation of the leading part of the spectrum pertinent to multi-dimensional eigenvalue problems (EVP) governing instability of flows with two or three inhomogeneous spatial directions. In the context of the numerical work presented in this thesis, the discretization of the spatial operator resulting from linearization of the Navier-Stokes equations around flows with two or three inhomogeneous spatial directions by variable-high-order stable finite-difference methods has permitted a speedup of four orders of magnitude in the solution of the corresponding two- and three-dimensional EVPs. This improvement of numerical performance has been achieved thanks to the high-sparsity level offered by the high-order finite-difference schemes employed for the discretization of the operators. This permitted use of efficient sparse linear algebra techniques without sacrificing accuracy and, consequently, solutions being obtained on typical workstations, as opposed to the previously employed supercomputers. Besides solution of the two- and three-dimensional EVPs of global linear instability, this development paved the way for the extension of the (linear and nonlinear) Parabolized Stability Equations (PSE) to analyze instability of flows which depend in a strongly-coupled inhomogeneous manner on two spatial directions and weakly on the third. Precisely the extensibility of the novel PSE-3D algorithm developed in the framework of the present thesis to study nonlinear flow instability permits transition prediction in flows of industrial interest, thus extending the classic PSE concept which has been successfully employed in the same context to boundary-layer type of flows over the last three decades. Typical examples of incompressible flows, the instability of which was analyzed in the present thesis without the need to resort to the restrictive assumptions used in the past, range from isolated vortices, and systems thereof, in which axial homogeneity is relaxed to consider viscous diffusion, as well as turbulent swirling jets, the instability of which is exploited in order to improve flame-holding properties of combustors. The instability of compressible subsonic and supersonic leading edge flows has been solved, and the wake of an isolated roughness element in a supersonic and hypersonic boundary-layer has also been analyzed with respect to its instability: excellent agreement with direct numerical simulation results has been obtained in all cases. Finally, instability analysis of Mach number 7 ow around an elliptic cone modeling the HIFiRE-5 flight test vehicle has unraveled flow instabilities near the minor-axis centerline, results comparing favorably with flight test predictions.

BiGlobal and point vortex methods for the instability analysis of wakes

To better understand destruction mechanisms of wake-vortices behind aircraft, the point vortex method for stability (inviscid) used by Crow is here compared with viscous modal global stability analysis of the linearized Navier-Stokes equations acting on a two-dimensional basic flow, i.e. BiGlobal stability analysis. The fact that the BiGlobal method is viscous, and uses a flnite área vortex model, gives rise to results somewhat different from the point vortex model. It adds more parameters to the problem, but is more realistic.

Spatial linear global instability analysis of the HIFiRE-5 elliptic cone model flow

The linear instability of the three-dimensional boundary-layer over the HIFiRE-5 flight test geometry, i.e. a rounded-tip 2:1 elliptic cone, at Mach 7, has been analyzed through spatial BiGlobal analysis, in a effort to understand transition and accurately predict local heat loads on next-generation ight vehicles. The results at an intermediate axial section of the cone, Re x = 8x10 5, show three different families of spatially amplied linear global modes, the attachment-line and cross- ow modes known from earlier analyses, and a new global mode, peaking in the vicinity of the minor axis of the cone, termed \center-line mode". We discover that a sequence of symmetric and anti-symmetric centerline modes exist and, for the basic ow at hand, are maximally amplied around F* = 130kHz. The wavenumbers and spatial distribution of amplitude functions of the centerline modes are documented

On linear instability mechanisms in incompressible open cavity flow

A theoretical study of linear global instability of incompressible flow over a rectangular spanwise-periodic open cavity in an unconfined domain is presented. Comparisons with the limited number of results available in the literature are shown. Subsequently, the parameter space is scanned in a systematic manner, varying Reynolds number, incoming boundary-layer thickness and length-to-depth aspect ratio. This permits documenting the neutral curves and leading eigenmode characteristics of this flow. Correlations constructed using the results obtained collapse all available theoretical data on the three-dimensional instabilities.

Instability Analysis of Incompressible Open Cavity Flows

El problema del flujo sobre una cavidad abierta ha sido estudiado en profundidad en la literatura, tanto por el interés académico del problema como por sus aplicaciones prácticas en gran variedad de problemas ingenieriles, como puede ser el alojamiento del tren de aterrizaje de aeronaves, o el depósito de agua de aviones contraincendios. Desde hace muchos a˜nos se estudian los distintos tipos de inestabilidades asociadas a este problema: los modos bidimensionales en la capa de cortadura, y los modos tridimensionales en el torbellino de recirculación principal dentro de la cavidad. En esta tesis se presenta un estudio paramétrico completo del límite incompresible del problema, empleando la herramienta de estabilidad lineal conocida como BiGlobal. Esta aproximación permite contemplar la estabilidad global del flujo, y obtener tanto la forma como las características de los modos propios del problema físico, sean estables o inestables. El estudio realizado permite caracterizar con gran detalle todos los modos relevantes, así como la envolvente de estabilidad en el espacio paramétrico del problema incompresible (Mach nulo, variación de Reynolds, espesor de capa límite incidente, relación altura/profundidad de la cavidad, y longitud característica de la perturbación en la dirección transversal). A la luz de los resultados obtenidos se proponen una serie de relaciones entre los parámetros y características de los modos principales, como por ejemplo entre el Reynolds crítico de un modo, y la longitud característica del mismo. Los resultados numéricos se contrastan con una campaña experimental, siendo la principal conclusión de dicha comparación que los modos lineales están presentes en el flujo real saturado, pero que existen diferencias notables en frecuencia entre las predicciones teóricas y los experimentos. Para intentar determinar la naturaleza de dichas diferencias se realiza una simulación numérica directa tridimensional, y se utiliza un algoritmo de DMD (descomposición dinámica de modos) para describir el proceso de saturación. ABSTRACT The problem of the flow over an open cavity has been studied in depth in the literature, both for being an interesting academical problem and due to the multitude of industrial applications, like the landing gear of aircraft, or the water deposit of firefighter airplanes. The different types of instabilities appearing in this flow studied in the literature are two: the two-dimensional shear layer modes, and the three-dimensional modes that appear in the main recirculating vortex inside the cavity. In this thesis a parametric study in the incompressible limit of the problem is presented, using the linear stability analysis known as BiGlobal. This approximation allows to obtain the global stability behaviour of the flow, and to capture both the morphological features and the characteristics of the eigenmodes of the physical problem, whether they are stable or unstable. The study presented here characterizes with great detail all the relevant eigenmodes, as well as the hypersurface of instability on the parameter space of the incompressible problem (Mach equal to zero, and variation of the Reynolds number, the incoming boundary layer thickness, the length to depth aspect ratio of the cavity and the spanwise length of the perturbation). The results allow to construct parametric relations between the characteristics of the leading eigenmodes and the parameters of the problem, like for example the one existing between the critical Reynolds number and its characteristic length. The numerical results presented here are compared with those of an experimental campaign, with the main conclusion of said comparison being that the linear eigenmode are present in the real saturated flow, albeit with some significant differences in the frequencies of the experiments and those predicted by the theory. To try to determine the nature of those differences a three-dimensional direct numerical simulation, analyzed with Dynamic Mode Decomposition algorithm, was used to describe the process of saturation.

On the Use of Matrix-Free Shift-Invert Strategies For Global Flow Instability Analysis

A novel time-stepping shift-invert algorithm for linear stability analysis of laminar flows in complex geometries is presented. This method, based on a Krylov subspace iteration, enables the solution of complex non-symmetric eigenvalue problems in a matrix-free framework. Validations and comparisons to the classical exponential method have been performed in three different cases: (i) stenotic flow, (ii) backward-facing step and (iii) lid-driven swirling flow. Results show that this new approach speeds up the required Krylov subspace iterations and has the capability of converging to specific parts of the global spectrum. It is shown that, although the exponential method remains the method of choice if leading eigenvalues are sought, the performance of the present method could be dramatically improved with the use of a preconditioner. In addition, as opposed to other methods, this strategy can be directly applied to any time-stepper, regardless of the temporal or spatial discretization of the latter.

On the role of global flow instability analysis in closed loop flow control

Control of linear flow instabilities has been demonstrated to be an effective theoretical flow control methodology, capable of modifying transitional flows on canonical geometries such as the plane channel and the flat-plate boundary layer. Extending the well-developed theoretical flow control techniques to flows over or through complex geometries requires addressing the issue of efficient capturing of the leading members of the global eigenspectrum pertinent to such flows. The present contribution describes state-of-the-art modal global instability analysis methodologies recently developed in our group, based on matrix formation and time-stepping, respectively. The relative performance of these algorithms is assessed on the recovery of BiGlobal and TriGlobal eigenspectra in the spanwise periodic and the cubic lid-driven cavity, respectively; the adjoint eigenspectrum in the latter flow is recovered for the first time. For three-dimensional flows without any homogeneous spatial direction, the time-stepping methodology was found to outperform the matrix-forming approach and permit recovering the leading TriGlobal eigenmodes in an three-dimensional open cavity of aspect ratio L : D : W = 5 : 1 : 1; theoretical flow control of this configuration is underway.

On the role of global flow instability analysis in closed loop flow control

Control of linear flow instabilities has been demonstrated to be an effective theoretical flow control methodology, capable of modifying transitional flow on canonical geometries such as the plane channel and the flat-plate boundary layer.

Linear instability analysis of incompressible flow over a cuboid cavity

Direct numerical simulations are performed to analyze the three-dimensional instability of flows over three-dimensional cavities. The flow structures at different Reynolds numbers are investigated by using the spectral-element solver nek5000. As the Reynolds number increasing, the lateral wall effects become more important, the recirculation zone shrinks, the front vortex increases and the flow structure inside of the cavity becomes more complex. Results show that the flow bifurcates from a steady state to an oscillatory regime beyond a value of Reynolds number Re = 1100.

Infrared thermography and shoulder pain in wheelchair users = Termografía infrarroja y dolor de hombro en usuarios de sillas de ruedas

Las personas que usan la silla de ruedas como su forma de movilidad prioritaria presentan una elevada incidencia (73%) de dolor de hombro debido al sobreuso y al movimiento repetitivo de la propulsión. Existen numerosos métodos de diagnóstico para la detección de las patologías del hombro, sin embargo la literatura reclama la necesidad de un test no invasivo y fiable, y sugiere la termografía como una técnica adecuada para evaluar el dolor articular. La termografía infrarroja (IRT) proporciona información acerca de los procesos fisiológicos a través del estudio de las distribuciones de la temperatura de la piel. Debido a la alta correlación entre ambos lados corporales, las asimetrías térmicas entre flancos contralaterales son una buena indicación de patologías o disfunciones físicas subyacentes. La fiabilidad de la IRT ha sido estudiada con anterioridad en sujetos sanos, pero nunca en usuarios de sillas de ruedas. Las características especiales de la población con discapacidad (problemas de sudoración y termorregulación, distribución sanguínea o medicación), hacen necesario estudiar los factores que afectan a la aplicación de la IRT en usuarios de sillas de ruedas. La bibliografía discrepa en cuanto a los beneficios o daños resultantes de la práctica de la actividad física en las lesiones de hombro por sobreuso en usuarios de sillas de ruedas. Recientes resultados apuntan a un aumento del riesgo de rotura del manguito rotador en personas con paraplejia que practican deportes con elevación del brazo por encima de la cabeza. Debido a esta falta de acuerdo en la literatura, surge la necesidad de analizar el perfil termográfico en usuarios de sillas de ruedas sedentarios y deportistas y su relación con el dolor de hombro. Hasta la fecha sólo se han publicado estudios termográficos durante el ejercicio en sujetos sanos. Un mayor entendimiento de la respuesta termográfica al ejercicio en silla de ruedas en relación al dolor de hombro clarificará su aparición y desarrollo y permitirá una apropiada intervención. El primer estudio demuestra que la fiabilidad de la IRT en usuarios de sillas de ruedas varía dependiendo de las zonas analizadas, y corrobora que la IRT es una técnica no invasiva, de no contacto, que permite medir la temperatura de la piel, y con la cual avanzar en la investigación en usuarios de sillas de ruedas. El segundo estudio proporciona un perfil de temperatura para usuarios de sillas de ruedas. Los sujetos no deportistas presentaron mayores asimetrías entre lados corporales que los sedentarios, y ambos obtuvieron superiores asimetrías que los sujetos sin discapacidad reportados en la literatura. Los no deportistas también presentaron resultados más elevados en el cuestionario de dolor de hombro. El área con mayores asimetrías térmicas fue hombro. En deportistas, algunas regiones de interés (ROIs) se relacionaron con el dolor de hombro. Estos resultados ayudan a entender el mapa térmico en usuarios de sillas de ruedas. El último estudio referente a la evaluación de la temperatura de la piel en usuarios de sillas de ruedas en ejercicio, reportó diferencias significativas entre la temperatura de la piel antes del test y 10 minutos después del test de propulsión de silla de ruedas, en 12 ROIs; y entre el post-test y 10 minutos después del test en la mayoría de las ROIs. Estas diferencias se vieron atenuadas cuando se compararon las asimetrías antes y después del test. La temperatura de la piel tendió a disminuir inmediatamente después completar el ejercicio, e incrementar significativamente 10 minutos después. El análisis de las asimetrías vs dolor de hombro reveló relaciones significativas negativas en 5 de las 26 ROIs. No se encontraron correlaciones significativas entre las variables de propulsión y el cuestionario de dolor de hombro. Todas las variables cinemáticas correlacionaron significativamente con las asimetrías en múltiples ROIs. Estos resultados indican que los deportistas en sillas de ruedas exhiben una capacidad similar de producir calor que los deportistas sin discapacidad; no obstante, su patrón térmico es más característico de ejercicios prolongados que de esfuerzos breves. Este trabajo contribuye al conocimiento de la termorregulación en usuarios de sillas de ruedas durante el ejercicio, y aporta información relevante para programas deportivos y de rehabilitación. ABSTRACT Individuals who use wheelchairs as their main means of mobility have a high incidence (73%) of shoulder pain (SP) owing to overuse and repetitive propulsion movement. There are numerous diagnostic methods for the detection of shoulder pathologies, however the literature claims that a noninvasive accurate test to properly assess shoulder pain would be necessary, and suggests thermography as a suitable technique for joint pain evaluation. Infrared thermography (IRT) provides information about physiological processes by studying the skin temperature (Tsk) distributions. Due to the high correlation of skin temperature between both sides of the body, thermal asymmetries between contralateral flanks are an indicator of underlying pathologies or physical dysfunctions. The reliability of infrared thermography has been studied in healthy subjects but there are no studies that have analyzed the reliability of IRT in wheelchair users (WCUs). The special characteristics of people with disabilities (sweating and thermoregulation problems, or blood distribution) make it necessary to study the factors affecting the application of IRT in WCUs. Discrepant reports exist on the benefits of, or damage resulting from, physical exercise and the relationship to shoulder overuse injuries in WCUs. Recent findings have found that overhead sports increase the risk of rotator cuff tears in wheelchair patients with paraplegia. Since there is no agreement in the literature, the thermographic profile of wheelchair athletes and nonathletes and its relation with shoulder pain should also be analysed. Infrared thermographic studies during exercise have been carried out only with able-bodied population at present. The understanding of the thermographic response to wheelchair exercise in relation to shoulder pain will offer an insight into the development of shoulder pain, which is necessary for appropriate interventions. The first study presented in this thesis demonstrates that the reliability of IRT in WCUs varies depending on the areas of the body that are analyzed. Moreover, it corroborates that IRT is a noninvasive and noncontact technique that allows the measurement of Tsk, which will allow for advances to be made in research concerned with WCUs. The second study provides a thermal profile of WCUs. Nonathletic subjects presented higher side-to-side skin temperature differences (ΔTsk) than athletes, and both had greater ΔTsk than the able-bodied results that have been published in the literature. Nonathletes also revealed larger Wheelchair Users Shoulder Pain Index (WUSPI) score than athletes. The shoulder region of interest (ROI) was the area with the highest ΔTsk of the regions measured. The analysis of the athletes’ Tsk showed that some ROIs are related to shoulder pain. These findings help to understand the thermal map in WCUs. Finally, the third study evaluated the thermal response of WCUs in exercise. There were significant differences in Tsk between the pre-test and the post-10 min in 12 ROIs, and between the post-test and the post-10 in most of the ROIs. These differences were attenuated when the ΔTsk was compared before and after exercise. Skin temperature tended to initially decrease immediately after the test, followed by a significant increase at 10 minutes after completing the exercise. The ΔTsk versus shoulder pain analysis yielded significant inverse relationships in 5 of the 26 ROIs. No significant correlations between propulsion variables and the results of the WUSPI questionnaire were found. All kinematic variables were significantly correlated with the temperature asymmetries in multiple ROIs. These results present indications that high performance wheelchair athletes exhibit similar capacity of heat production to able-bodied population; however, they presented a thermal pattern more characteristic of a prolonged exercise rather than brief exercise. This work contributes to improve the understanding about temperature changes in wheelchair athletes during exercise and provides implications to the sports and rehabilitation programs.

Global instability analysis and control of three-dimensional long open cavity flow

The three-dimensional wall-bounded open cavity may be considered as a simplified geometry found in industrial applications such as leading gear or slotted flats on the airplane. Understanding the three-dimensional complex flow structure that surrounds this particular geometry is therefore of major industrial interest. At the light of the remarkable former investigations in this kind of flows, enough evidences suggest that the lateral walls have a great influence on the flow features and hence on their instability modes. Nevertheless, even though there is a large body of literature on cavity flows, most of them are based on the assumption that the flow is two-dimensional and spanwise-periodic. The flow over realistic open cavity should be considered. This thesis presents an investigation of three-dimensional wall-bounded open cavity with geometric ratio 6:2:1. To this aim, three-dimensional Direct Numerical Simulation (DNS) and global linear instability have been performed. Linear instability analysis reveals that the onset of the first instability in this open cavity is around Recr 1080. The three-dimensional shear layer mode with a complex structure is shown to be the most unstable mode. I t is noteworthy that the flow pattern of this high-frequency shear layer mode is similar to the observed unstable oscillations in supercritical unstable case. DNS of the cavity flow carried out at different Reynolds number from steady state until a nonlinear saturated state is obtained. The comparison of time histories of kinetic energy presents a clearly dominant energetic mode which shifts between low-frequency and highfrequency oscillation. A complete flow patterns from subcritical cases to supercritical case has been put in evidence. The flow structure at the supercritical case Re=1100 resembles typical wake-shedding instability oscillations with a lateral motion existed in the subcritical cases. Also, This flow pattern is similar to the observations in experiments. In order to validate the linear instability analysis results, the topology of the composite flow fields reconstructed by linear superposition of a three-dimensional base flow and its leading three-dimensional global eigenmodes has been studied. The instantaneous wall streamlines of those composited flows display distinguish influence region of each eigenmode. Attention has been focused on the leading high-frequency shear layer mode; the composite flow fields have been fully recognized with respect to the downstream wave shedding. The three-dimensional shear layer mode is shown to give rise to a typical wake-shedding instability with a lateral motions occurring downstream which is in good agreement with the experiment results. Moreover, the spanwise-periodic, open cavity with the same length to depth ratio has been also studied. The most unstable linear mode is different from the real three-dimensional cavity flow, because of the existence of the side walls. Structure sensitivity of the unstable global mode is analyzed in the flow control context. The adjoint-based sensitivity analysis has been employed to localized the receptivity region, where the flow is more sensible to momentum forcing and mass injection. Because of the non-normality of the linearized Navier-Stokes equations, the direct and adjoint field has a large spatial separation. The strongest sensitivity region is locate in the upstream lip of the three-dimensional cavity. This numerical finding is in agreement with experimental observations. Finally, a prototype of passive flow control strategy is applied.

Transient excess of MYC activity can elicit genomic instability and tumorigenesis

Overexpression of the MYC protooncogene has been implicated in the genesis of diverse human tumors. Tumorigenesis induced by MYC has been attributed to sustained effects on proliferation and differentiation. Here we report that MYC may also contribute to tumorigenesis by destabilizing the cellular genome. A transient excess of MYC activity increased tumorigenicity of Rat1A cells by at least 50-fold. The increase persisted for >30 days after the return of MYC activity to normal levels. The brief surfeit of MYC activity was accompanied by evidence of genomic instability, including karyotypic abnormalities, gene amplification, and hypersensitivity to DNA-damaging agents. MYC also induced genomic destabilization in normal human fibroblasts, although these cells did not become tumorigenic. Stimulation of Rat1A cells with MYC accelerated their passage through G1/S. Moreover, MYC could force normal human fibroblasts to transit G1 and S after treatment with N-(phosphonoacetyl)-l-aspartate (PALA) at concentrations that normally lead to arrest in S phase by checkpoint mechanisms. Instead, the cells subsequently appeared to arrest in G2. We suggest that the accelerated passage through G1 was mutagenic but that the effect of MYC permitted a checkpoint response only after G2 had been reached. Thus, MYC may contribute to tumorigenesis through a dominant mutator effect.

Instability of signaling resolution models of parent–offspring conflict

Recent signaling resolution models of parent–offspring conflict have provided an important framework for theoretical and empirical studies of communication and parental care. According to these models, signaling of need is stabilized by its cost. However, our computer simulations of the evolutionary dynamics of chick begging and parental investment show that in Godfray’s model the signaling equilibrium is evolutionarily unstable: populations that start at the signaling equilibrium quickly depart from it. Furthermore, the signaling and nonsignaling equilibria are linked by a continuum of equilibria where chicks above a certain condition do not signal and we show that, contrary to intuition, fitness increases monotonically as the proportion of young that signal decreases. This result forces us to reconsider much of the current literature on signaling of need and highlights the need to investigate the evolutionary stability of signaling equilibria based on the handicap principle.

Induction of microsatellite instability by oxidative DNA damage

Instability of repetitive sequences, both in intronic sequences and within coding regions, has been demonstrated to be a hallmark of genomic instability in human cancer. Understanding how these mutational events arise may provide an opportunity for prevention or early intervention in cancer development. To study the source of this instability, we have identified a region of the β-lactamase gene that is tolerant to the insertion of fragments of exogenous DNA as large as 1,614 bp with minimal loss of enzyme activity, as determined by antibiotic resistance. Fragments inserted out-of-frame render Escherichia coli sensitive to antibiotic, and compensatory frameshift mutations that restore the reading frame of β-lactamase can be selected on the basis of antibiotic resistance. We have utilized this site to insert a synthetic microsatellite sequence within the β-lactamase gene and selected for mutations yielding frameshifts. This assay provides for detection of one frameshift mutation in a background of 106 wild-type sequences. Mismatch repair deficiency increased the observed frameshift frequency ≈300-fold. Exposure of plasmid containing microsatellite sequences to hydrogen peroxide resulted in frameshift mutations that were localized exclusively to the microsatellite sequences, whereas DNA damage by UV or N-methyl-N′-nitro-N-nitrosoguanidine did not result in enhanced mutagenesis. We postulate that in tumor cells, endogenous production of oxygen free radicals may be a major factor in promoting instability of microsatellite sequences. This β-lactamase assay may provide a sensitive methodology for the detection and quantitation of mutations associated with the development of cancer.

The human XRCC9 gene corrects chromosomal instability and mutagen sensitivities in CHO UV40 cells

The Chinese hamster ovary (CHO) mutant UV40 cell line is hypersensitive to UV and ionizing radiation, simple alkylating agents, and DNA cross-linking agents. The mutant cells also have a high level of spontaneous chromosomal aberrations and 3-fold elevated sister chromatid exchange. We cloned and sequenced a human cDNA, designated XRCC9, that partially corrected the hypersensitivity of UV40 to mitomycin C, cisplatin, ethyl methanesulfonate, UV, and γ-radiation. The spontaneous chromosomal aberrations in XRCC9 cDNA transformants were almost fully corrected whereas sister chromatid exchanges were unchanged. The XRCC9 genomic sequence was cloned and mapped to chromosome 9p13. The translated XRCC9 sequence of 622 amino acids has no similarity with known proteins. The 2.5-kb XRCC9 mRNA seen in the parental cells was undetectable in UV40 cells. The mRNA levels in testis were up to 10-fold higher compared with other human tissues and up to 100-fold higher compared with other baboon tissues. XRCC9 is a candidate tumor suppressor gene that might operate in a postreplication repair or a cell cycle checkpoint function.