Expanding the Toolkit for Synthetic Biology: Frameworks for Native-like Non-natural Gene Circuits

Synthetic biology combines biological parts from different sources in order to engineer non-native, functional systems. While there is a lot of potential for synthetic biology to revolutionize processes, such as the production of pharmaceuticals, engineering synthetic systems has been challenging. It is oftentimes necessary to explore a large design space to balance the levels of interacting components in the circuit. There are also times where it is desirable to incorporate enzymes that have non-biological functions into a synthetic circuit. Tuning the levels of different components, however, is often restricted to a fixed operating point, and this makes synthetic systems sensitive to changes in the environment. Natural systems are able to respond dynamically to a changing environment by obtaining information relevant to the function of the circuit. This work addresses these problems by establishing frameworks and mechanisms that allow synthetic circuits to communicate with the environment, maintain fixed ratios between components, and potentially add new parts that are outside the realm of current biological function. These frameworks provide a way for synthetic circuits to behave more like natural circuits by enabling a dynamic response, and provide a systematic and rational way to search design space to an experimentally tractable size where likely solutions exist. We hope that the contributions described below will aid in allowing synthetic biology to realize its potential.

Predation and selection in copepod populations of Alpine waters [Translation from: Bollettino di Zoologia 21 541-545, 1954]

Zooplankton was studied in four alpine lakes in Switzerland, France and Italy. The presence the presence of the invertebrate predator Heterocope in three lakes was stated. It is then discussed why in three of these four lakes, the copepod Arctodiaptomus denticornis is present in the absence of Arctodiaptomus bacillifer, and vice versa respectively in the second and first parts of the lacustrine summer.

Investigations in the theory of electromagnetic scattering from expanding dielectric obstacles

An equation for the reflection which results when an expanding dielectric slab scatters normally incident plane electromagnetic waves is derived using the invariant imbedding concept. The equation is solved approximately and the character of the solution is investigated. Also, an equation for the radiation transmitted through such a slab is similarly obtained. An alternative formulation of the slab problem is presented which is applicable to the analogous problem in spherical geometry. The form of an equation for the modal reflections from a nonrelativistically expanding sphere is obtained and some salient features of the solution are described. In all cases the material is assumed to be a nondispersive, nonmagnetic dielectric whose rest frame properties are slowly varying.

Robust Control of Evolutionary Dynamics

The application of principles from evolutionary biology has long been used to gain new insights into the progression and clinical control of both infectious diseases and neoplasms. This iterative evolutionary process consists of expansion, diversification and selection within an adaptive landscape - species are subject to random genetic or epigenetic alterations that result in variations; genetic information is inherited through asexual reproduction and strong selective pressures such as therapeutic intervention can lead to the adaptation and expansion of resistant variants. These principles lie at the center of modern evolutionary synthesis and constitute the primary reasons for the development of resistance and therapeutic failure, but also provide a framework that allows for more effective control.

A model system for studying the evolution of resistance and control of therapeutic failure is the treatment of chronic HIV-1 infection by broadly neutralizing antibody (bNAb) therapy. A relatively recent discovery is that a minority of HIV-infected individuals can produce broadly neutralizing antibodies, that is, antibodies that inhibit infection by many strains of HIV. Passive transfer of human antibodies for the prevention and treatment of HIV-1 infection is increasingly being considered as an alternative to a conventional vaccine. However, recent evolution studies have uncovered that antibody treatment can exert selective pressure on virus that results in the rapid evolution of resistance. In certain cases, complete resistance to an antibody is conferred with a single amino acid substitution on the viral envelope of HIV.

The challenges in uncovering resistance mechanisms and designing effective combination strategies to control evolutionary processes and prevent therapeutic failure apply more broadly. We are motivated by two questions: Can we predict the evolution to resistance by characterizing genetic alterations that contribute to modified phenotypic fitness? Given an evolutionary landscape and a set of candidate therapies, can we computationally synthesize treatment strategies that control evolution to resistance?

To address the first question, we propose a mathematical framework to reason about evolutionary dynamics of HIV from computationally derived Gibbs energy fitness landscapes -- expanding the theoretical concept of an evolutionary landscape originally conceived by Sewall Wright to a computable, quantifiable, multidimensional, structurally defined fitness surface upon which to study complex HIV evolutionary outcomes.

To design combination treatment strategies that control evolution to resistance, we propose a methodology that solves for optimal combinations and concentrations of candidate therapies, and allows for the ability to quantifiably explore tradeoffs in treatment design, such as limiting the number of candidate therapies in the combination, dosage constraints and robustness to error. Our algorithm is based on the application of recent results in optimal control to an HIV evolutionary dynamics model and is constructed from experimentally derived antibody resistant phenotypes and their single antibody pharmacodynamics. This method represents a first step towards integrating principled engineering techniques with an experimentally based mathematical model in the rational design of combination treatment strategies and offers predictive understanding of the effects of combination therapies of evolutionary dynamics and resistance of HIV. Preliminary in vitro studies suggest that the combination antibody therapies predicted by our algorithm can neutralize heterogeneous viral populations despite containing resistant mutations.

The fish populations of the River Avon, Hampshire, downstream of Salisbury, in 1987

The objective of this study is to describe the present statu s of the coarse fish populations of the River Avon, Hampshire. The study arose from a widespread concern among anglers and riparian owners that fish numbers had decreased since the early 1970's, and from a scarcity of accurate information on population densities on which to assess this complaint. It was realised at the outset that, without such information from earlier years, the survey would not reveal the extent of any decline in the fisheries. The result s of the survey are compared with available data from other rivers , chiefly the nearby R. Frome and R. Stour. They may be used to assess any futur e changes in population densities and growth rates, and to indicate promising areas for future research. The data were collected from a quantitative survey of twelve river sections along the fifty kilometres of river between Salisbury and Sopley, and a qualitative survey of three weir pools within the same area from 14 September to 9 October, 1987.

Variation in bacterial populations in time and space

Obtaining a reliable estimate of the bacterial population is one of the main problems facing the bacterial ecologist. The author discusses the various methods available and concludes that the observed variability in bacterial populations depends on the sampling interval used.

The impact of Cow Green Reservoir on invertebrate populations in the River Tees

Cow Green is a new reservoir situated in Pennine moorland. It has an area of 312 ha, a capacity of 40 . 9 x 10 SUP-6 m SUP-3 and a maximum depth of 22 . 8 m. The function of the reservoir is to regulate flow in the River Tees to provide industrial Teesside with sufficient water during the dry spells. Invertebrate studies were carried out in the Tees to monitor changes resulting from the construction of the reservoir both in the flooded basin and below the dam. The overall effect of the reservoir on the Tees has been to increase the numbers and biomass of certain taxa, but generally not at the expense of previous fauna. Some of the positive effects, ie. increase in number and biomass, and maintenance of faunal diversity, may in part be attributable to the presence of the rapids and waterfall. Turbulence resulting from this rapid flow over heterogeneous bottom is sufficient to prevent clogging of interstitial spaces by silt and to maintain the variety of ecological niches necessary for a diverse fauna.

Effects of Cow Green Reservoir upon downstream fish populations

Post impoundment observations on the fish populations of the River Tees, downstream from the Cow Green Reservoir were made between 1971 and 1980. Both bullhead (Cottus gobio L.) and brown trout (Salmo trutta L.) population densities increased in the river after impoundment but changes in growth rate were small. Investigations into the stomach contents of the two species reflects the results of other work on increases of benthos in the river. Following regulation there was an increase in the quantity of Ephemerella ignita found in trout stomachs while in the bullhead, regulation caused an increase in the importance of Mollusca and a decrease in importance of Ephemeroptera and Plecoptera.

A-SCALA: an age-structured statistical catch-at-length analysis for assessing tuna stocks in the eastern tropical Pacific Ocean

English: We describe an age-structured statistical catch-at-length analysis (A-SCALA) based on the MULTIFAN-CL model of Fournier et al. (1998). The analysis is applied independently to both the yellowfin and the bigeye tuna populations of the eastern Pacific Ocean (EPO). We model the populations from 1975 to 1999, based on quarterly time steps. Only a single stock for each species is assumed for each analysis, but multiple fisheries that are spatially separate are modeled to allow for spatial differences in catchability and selectivity. The analysis allows for error in the effort-fishing mortality relationship, temporal trends in catchability, temporal variation in recruitment, relationships between the environment and recruitment and between the environment and catchability, and differences in selectivity and catchability among fisheries. The model is fit to total catch data and proportional catch-at-length data conditioned on effort. The A-SCALA method is a statistical approach, and therefore recognizes that the data collected from the fishery do not perfectly represent the population. Also, there is uncertainty in our knowledge about the dynamics of the system and uncertainty about how the observed data relate to the real population. The use of likelihood functions allow us to model the uncertainty in the data collected from the population, and the inclusion of estimable process error allows us to model the uncertainties in the dynamics of the system. The statistical approach allows for the calculation of confidence intervals and the testing of hypotheses. We use a Bayesian version of the maximum likelihood framework that includes distributional constraints on temporal variation in recruitment, the effort-fishing mortality relationship, and catchability. Curvature penalties for selectivity parameters and penalties on extreme fishing mortality rates are also included in the objective function. The mode of the joint posterior distribution is used as an estimate of the model parameters. Confidence intervals are calculated using the normal approximation method. It should be noted that the estimation method includes constraints and priors and therefore the confidence intervals are different from traditionally calculated confidence intervals. Management reference points are calculated, and forward projections are carried out to provide advice for making management decisions for the yellowfin and bigeye populations. Spanish: Describimos un análisis estadístico de captura a talla estructurado por edad, A-SCALA (del inglés age-structured statistical catch-at-length analysis), basado en el modelo MULTIFAN- CL de Fournier et al. (1998). Se aplica el análisis independientemente a las poblaciones de atunes aleta amarilla y patudo del Océano Pacífico oriental (OPO). Modelamos las poblaciones de 1975 a 1999, en pasos trimestrales. Se supone solamente una sola población para cada especie para cada análisis, pero se modelan pesquerías múltiples espacialmente separadas para tomar en cuenta diferencias espaciales en la capturabilidad y selectividad. El análisis toma en cuenta error en la relación esfuerzo-mortalidad por pesca, tendencias temporales en la capturabilidad, variación temporal en el reclutamiento, relaciones entre el medio ambiente y el reclutamiento y entre el medio ambiente y la capturabilidad, y diferencias en selectividad y capturabilidad entre pesquerías. Se ajusta el modelo a datos de captura total y a datos de captura a talla proporcional condicionados sobre esfuerzo. El método A-SCALA es un enfoque estadístico, y reconoce por lo tanto que los datos obtenidos de la pesca no representan la población perfectamente. Además, hay incertidumbre en nuestros conocimientos de la dinámica del sistema e incertidumbre sobre la relación entre los datos observados y la población real. El uso de funciones de verosimilitud nos permite modelar la incertidumbre en los datos obtenidos de la población, y la inclusión de un error de proceso estimable nos permite modelar las incertidumbres en la dinámica del sistema. El enfoque estadístico permite calcular intervalos de confianza y comprobar hipótesis. Usamos una versión bayesiana del marco de verosimilitud máxima que incluye constreñimientos distribucionales sobre la variación temporal en el reclutamiento, la relación esfuerzo-mortalidad por pesca, y la capturabilidad. Se incluyen también en la función objetivo penalidades por curvatura para los parámetros de selectividad y penalidades por tasas extremas de mortalidad por pesca. Se usa la moda de la distribución posterior conjunta como estimación de los parámetros del modelo. Se calculan los intervalos de confianza usando el método de aproximación normal. Cabe destacar que el método de estimación incluye constreñimientos y distribuciones previas y por lo tanto los intervalos de confianza son diferentes de los intervalos de confianza calculados de forma tradicional. Se calculan puntos de referencia para el ordenamiento, y se realizan proyecciones a futuro para asesorar la toma de decisiones para el ordenamiento de las poblaciones de aleta amarilla y patudo.

Modelling Lipid Competition Dynamics in Heterogeneous Protocell Populations

Recent experimental work in the field of synthetic protocell biology has shown that prebiotic vesicles are able to 'steal' lipids from each other. This phenomenon is driven purely by asymmetries in the physical state or composition of the vesicle membranes, and, when lipid resource is limited, translates directly into competition amongst the vesicles. Such a scenario is interesting from an origins of life perspective because a rudimentary form of cell-level selection emerges. To sharpen intuition about possible mechanisms underlying this behaviour, experimental work must be complemented with theoretical modelling. The aim of this paper is to provide a coarse-grain mathematical model of protocell lipid competition. Our model is capable of reproducing, often quantitatively, results from core experimental papers that reported distinct types vesicle competition. Additionally, we make some predictions untested in the lab, and develop a general numerical method for quickly solving the equilibrium point of a model vesicle population.

The juvenile salmonid populations of the River Lune catchment, 1981 to 1985

The aim of this study was to assess the status of the juvenile salmonid populations of the River Lune and its tributaries. There was special emphasis on juvenile salmon stocks in view of the implementation of a net limitation order in 1980. The number of licensed instruments were reduced as follows: (1) Drift, hang or whammel nets - from 12 to 10 (2) Draft or seine nets - from 3 to 1 (3) Heave or haaf nets - from 4 6 to 2 6 For the purpose of this report, the River Lune system has been divided into 13 subcatchments and these are examined with a view to detecting any trends in the data such as subcatchment productivity, partitioning between salmon and trout nursery streams, and whether the restrictions on salmon fishing had any discernible effects on juvenile salmon productivity. The effect of flow and instream obstructions on salmonid densities are also investigated as these have been shown to be important factors affecting abundance. Throughout the study period a programme of enhancement stocking took place primarily with salmon ova and fry. The possible impact of this on the results of the surveys has been assessed. The distribution and abundance of juvenile salmon was found to be relatively consistent in each of the 13 subcatchments studied over the 1981-1985 period.