One of the largest problems facing modern medicine is drug resistance. Many classes of drugs can be rendered ineffective if their target is able to acquire beneficial mutations. While this is an excellent showcase of the power of evolution, it necessitates the development of increasingly stronger drugs to combat resistant pathogens. Not only is this strategy costly and time consuming, it is also unsustainable. To contend with this problem, many multi-drug treatment strategies are being explored. Previous studies have shown that resistance to some drug combinations is not possible, for example, resistance to a common antifungal drug, fluconazole, seems impossible in the presence of radicicol. We believe that in order to understand the viability of multi-drug strategies in combating drug resistance, we must understand the full spectrum of resistance mutations that an organism can develop, not just the most common ones. It is possible that rare mutations exist that are resistant to both drugs. Knowing the frequency of such mutations is important for making predictions about how problematic they will be when multi-drug strategies are used to treat human disease. This experiment aims to expand on previous research on the evolution of drug resistance in S. cerevisiae by using molecular barcodes to track ~100,000 evolving lineages simultaneously. The barcoded cells were evolved with serial transfers for seven weeks (200 generations) in three concentrations of the antifungal Fluconazole, three concentrations of the Hsp90 inhibitor Radicicol, and in four combinations of Fluconazole and Radicicol. Sequencing data was used to track barcode frequencies over the course of the evolution, allowing us to observe resistant lineages as they rise and quantify differences in resistance evolution across the different conditions. We were able to successfully observe over 100,000 replicates simultaneously, revealing many adaptive lineages in all conditions. Our results also show clear differences across drug concentrations and combinations, with the highest drug concentrations exhibiting distinct behaviors.

Stephen Jay Gould studied snail fossils and worked at Harvard University in Cambridge, Massachusetts during the latter half of the twentieth century. He contributed to philosophical, historical, and scientific ideas in paleontology, evolutionary theory, and developmental biology. Gould, with Niles Eldredge, proposed the theory of punctuated equilibrium, a view of evolution by which species undergo long periods of stasis followed by rapid changes over relatively short periods instead of continually accumulating slow changes over millions of years. In his 1977 book, Ontogeny and Phylogeny, Gould reconstructed a history of developmental biology and stressed the importance of development to evolutionary biology. In a 1979 paper coauthored with Richard Lewontin, Gould and Lewonitn criticized many evolutionary bioligists for relying solely on adaptive evolution as an explanation for morphological change, and for failing to consider other explanations, such as developmental constraints.

David Starr Jordan studied fish and promoted eugenics in the US during the late nineteenth and early twentieth centuries. In his work, he embraced Charles Darwin s theory of evolution and described the importance of embryology in tracing phylogenic relationships. In 1891, he became the president of Stanford University in Stanford, California. Jordan condemned war and promoted conservationist causes for the California wilderness, and he advocated for the eugenic sterilization of thousands of Americans. Like many American eugenicists of the early twentieth century, Jordan combined ideas of Mendelian genetics and of Darwinian natural selection to form a basis for limiting or encouraging reproduction in certain individuals and groups based on their perceived hereditary fitness. Like other eugenicists, Jordan s attempt to control the reproductive fate of entire populations marked an episode in the history of reproduction and biology in which its concepts increasingly influenced the social and cultural contexts.

Georges Cuvier, baptized Georges Jean-Leopold Nicolas-Frederic Cuvier, was a professor of anatomy at the National Museum of Natural History in Paris, France, through the late eighteenth and early nineteenth centuries. Scholars recognize Cuvier as a founder of modern comparative anatomy, and as an important contributor to vertebrate paleontology and geology. Cuvier studied the form and function of animal anatomy, writing four volumes on quadruped fossils and co-writing eleven volumes on the natural history of fish with Achille Valenciennes. Moreover, Cuvier constructed a system of classification based on specific and well-articulated principles to help anatomists classify animal taxa. Cuvier had public debate in 1830 with Etienne Geoffroy Saint-Hilaire, a dispute centered on whether form or function matters most for the study of anatomy and whether the transmutation of organic forms can occur over time. Cuvier's opinions influenced the development of biology in France, and his arguments against transmutation of types influenced the reception of Charles Darwin's theory of evolution by natural selection among many French naturalists.

Etienne Geoffroy Saint-Hilaire, commonly known as Geoffroy, studied animals, their anatomy and their embryos, and teratogens at the National Museum of Natural History in Paris, France in the eighteenth and nineteenth centuries. Geoffroy also helped develop several specialized fields in the life sciences, including experimental embryology. In his efforts to experimentally demonstrate the theory of recapitulation, Geoffroy developed techniques to intervene in the growth of embryos to see whether they would develop into different kinds of organisms. Moreover, Geoffroy emphasized the concept of l'unite de composition (the unity of plan). Geoffroy disputed in 1830 with Georges Cuvier over whether form or function matters most for the study of anatomy and whether the transformation of organic forms can occur over time. Geoffroy's conceptual contributions, as well as his experimental research, influenced embryological research on animal morphology and teratogens, and later the field of evolutionary paleontology.

In his essay Evolution and Tinkering, published in
Science in 1977, Francois Jacob argued that a common analogy
between the process of evolution by natural selection and the
methods of engineering is problematic. Instead, he proposed to
describe the process of evolution with the concept of
bricolage (tinkering). In this essay, Jacob did not deny the
importance of the mechanism of natural selection in shaping complex
adaptations. Instead, he maintained that the cumulative effects of
history on the evolution of life, made evident by molecular data,
provides an alternative account of the patterns depicting the
history of life on earth. Jacob's essay contributed to
genetic research in the late twentieth century that emphasized
certain types of topics in evolutionary and developmental biology,
such as genetic regulation, gene duplication events, and the genetic
program of embryonic development. It also proposed why, in future
research, biologists should expect to discover an underlying
similarity in the molecular structure of genomes, and that they
should expect to find many imperfections in evolutionary history
despite the influence of natural selection.

In 2008 researchers Daniel Warner and Richard Shine tested the Charnov-Bull model by conducting experiments on the Jacky dragon (Amphibolurus muricatus), in Australia. Their results showed that temperature-dependent sex determination(TSD) evolved in this species as an adaptation to fluctuating environmental temperatures. The Charnov-Bull model, proposed by Eric Charnov and James Bull in 1977, described the evolution of TSD, although the model was, for many years, untested. Many reptiles and some fish exhibit non-genetic sex determination, in which an embryos' environment can influence the sex of the adult organism. Environmental conditions such as humidity or population density can alter sex in some organisms, and a widespread form of non-genetic sex determination is temperature-dependent sex determination. TSD reveals how embryonic development can contribute to the evolution of physiological processes. Researchers have documented TSD in a wide range of species, and they continue to investigate how such a sex determining system has evolved.

Pathogenic drug resistance is a major global health concern. Thus, there is great interest in modeling the behavior of resistant mutations–how quickly they will rise in frequency within a population, and whether they come with fitness tradeoffs that can form the basis of treatment strategies. These models often depend on precise measurements of the relative fitness advantage (s) for each mutation and the strength of the fitness tradeoff that each mutation suffers in other contexts. Precisely quantifying s helps us create better, more accurate models of how mutants act in different treatment strategies. For example, P. falciparum acquires antimalarial drug resistance through a series of mutations to a single gene. Prior work in yeast expressing this P. falciparum gene demonstrated that mutations come with tradeoffs. Computational work has demonstrated the possibility of a treatment strategy which enriches for a particular resistant mutation that then makes the population grow poorly once the drug is removed. This treatment strategy requires knowledge of s and how it changes when multiple mutants are competing across various drug concentrations. Here, we precisely quantified s in varying drug concentrations for five resistant mutants, each of which provide varying degrees of drug resistance to antimalarial drugs. DNA barcodes were used to label each strain, allowing the mutants to be pooled together for direct competition in different concentrations of drug. This will provide data that can make the models more accurate, potentially facilitating more effective drug treatments in the future.