Basal metabolic rate of endotherms can be modeled using heat-transfer principles and physiological concepts: reply to "can the basal metabolic rate of endotherms be explained by biophysical modeling?".

Our recent article (Roberts et al. 2010 ) proposes a mechanistic model for the relation between basal metabolic rate (BMR) and body mass (M) in mammals. The model is based on heat-transfer principles in the form of an equation for distributed heat generation within the body. The model can also be written in the form of the allometric equation BMR = aM(b), in which a is the coefficient of the mass term and b is the allometric exponent. The model generates two interesting results: it predicts that b takes the value 2/3, indicating that BMR is proportional to surface area in endotherms. It also provides an explanation of the physiological components that make up a, that is, respiratory heat loss, core-skin thermal conductance, and core-skin thermal gradient. Some of the ideas in our article have been questioned (Seymour and White 2011 ), and this is our response to those questions. We specifically address the following points: whether a heat-transfer model can explain the level of BMR in mammals, whether our test of the model is inadequate because it uses the same literature data that generated the values of the physiological variables, and whether geometry and empirical values combine to make a "coincidence" that makes the model only appear to conform to real processes.

A new model for the body size-metabolism relationship.

The allometric 3/4 power relation, initially used for describing the relation between mammalian basal metabolic rate and body size, is often used as a general model for organismal design. The use of allometric regression as a model has important limitations: it is not mechanistic, it combines all physiological variables into one correlate of body size, and it combines data from several physiological states. In reassessing the use of allometric equations, we first describe problems with their use in studies of organismal design and then use a formulation for distributed net heat production and temperature distribution within the body to derive an alternative equation for the relation between basal metabolism and body size. Tests of the heat flow equation against data reported in the literature indicate that it is an accurate estimator of basal metabolism under thermoneutral conditions and suggest that the allometric equation is a special case of this mechanistic and more general model. We propose that our method is more meaningful and widely applicable for thermoneutral conditions than is a purely allometric approach.

Emergence of ideal membrane cascades for downstream processing.

An algorithm is developed for describing ideal membrane cascades for fractionation of binary and pseudo-binary mixtures. It is shown that solvent management plays a key role in determining both purification and yield. Development of efficient diafilters is needed if membrane cascades are to achieve their full potential in competing with both chromatography and simulated moving bed operations in downstream processing of proteins. Such a replacement will also be important for fractionation of higher titers and larger substrates, such as plasmids, viruses, and even whole cells.

Identification of non-idealities in gel electrophoresis.

The molecular weight separation, which is the second dimension of two-dimensional gel electrophoresis, is studied quantitatively with the goal of improving positional predictability and reproducibility. Mathematical modeling of carrier electrolyte dynamics is used to track the progress of a stacking front as a function of coulombs passed. In all test cases, the front moves more slowly than predicted and shows both curvature and tilt. These systematic deviations from the model are found to be influenced by a variety of factors, including both design and operating features. These factors are largely explained, and suggestions are made for improvements.

Adsorption and desorption behavior of plasmid DNA on ion-exchange membranes. Effect of salt valence and compaction agents.

In this study, we report the effect of salt type and compaction agents on adsorption and desorption behavior of plasmid DNA on strong anion-exchange membranes. Both divalent cations and compaction agents are known to reduce the effective charge density of plasmid DNA in solution, and compaction agents decrease the radius of gyration of plasmids. Differences in the batch uptake adsorption of a 6.1 kilo base pair plasmid in solution with sodium and magnesium salts were observed at low ionic strengths. Recoveries at high salt conditions, however, were independent of the cation, and measured only 63-76%. Similarly, no improvement in recoveries were observed when using sulfate rather than chloride anions as displacers. The compaction agents, spermine and spermidine, showed no strong effect on the uptake adsorption, capacity, or recovery of three different-sized plasmids on membrane sheets. It is recommended that further efforts to improve plasmid recoveries from anion-exchange membranes focus on properties of the adsorbent surface.

Performance and scale-up of adsorptive membrane chromatography.

Separation efficiency and scalability of Pall Corporation's new Mustang stacked membrane chromatographic devices were investigated, using both the 10-ml and l(-1) models and comparing the responses of tracer pulses obtained for conventional and reverse-flow operation. Tracers included AMP, lysozyme, and thyroglobulin, which vary in relative molecular mass from less than 1000 up to 650000. Both devices showed marked insensitivity to tracer size and flow-rate and gave sharper peaks than would have been expected from conventional 15-microm bead packings. However, reverse-flow peaks were always significantly sharper than those for conventional operation, and the differences were ascribed primarily to non-uniform header residence times. Numerical simulations of the macroscopic flow confirmed that this was indeed the case. This problem was much less pronounced for the l(-1) device so scale-up is conservative.