Death or taxes? The political economy of sanitation expenditure in nineteenth-century Britain

This thesis consists of three papers studying the relationship between democratic reform, expenditure on sanitation public goods and mortality in Britain in the second half of the nineteenth century. During this period decisions over spending on critical public goods such as water supply and sewer systems were made by locally elected town councils, leading to extensive variation in the level of spending across the country. This dissertation uses new historical data to examine the political factors determining that variation, and the consequences for mortality rates.

The first substantive chapter describes the spread of government sanitation expenditure, and analyzes the factors that determined towns' willingness to invest. The results show the importance of towns' financial constraints, both in terms of the available tax base and access to borrowing, in limiting the level of expenditure. This suggests that greater involvement by Westminster could have been very effective in expediting sanitary investment. There is little evidence, however, that democratic reform was an important driver of greater expenditure.

Chapter 3 analyzes the effect of extending voting rights to the poor on government public goods spending. A simple model predicts that the rich and the poor will desire lower levels of public goods expenditure than the middle class, and so extensions of the right to vote to the poor will be associated with lower spending. This prediction is tested using plausibly exogenous variation in the extent of the franchise. The results strongly support the theoretical prediction: expenditure increased following relatively small extensions of the franchise, but fell once more than approximately 50% of the adult male population held the right to vote.

Chapter 4 tests whether the sanitary expenditure was effective in combating the high mortality rates following the Industrial Revolution. The results show that increases in urban expenditure on sanitation-water supply, sewer systems and streets-was extremely effective in reducing mortality from cholera and diarrhea.

A unified strategy to ent-kauranoid natural products : total syntheses of (–)-maoecrystal Z, (–)-trichorabdal A, and (–)-longikaurin E

The diterpenoid constituents of the Isodon plants have attracted reasearchers interested in both their chemical structures and biological properties for more than a half-century. In recent years, the isolations of new members displaying previously unprecedented ring systems and highly selective biological properties have piqued interest from the synthetic community in this class of natural products.

Reported herein is the first total synthesis of such a recently isolated diterpenoid, (–)-maoecrystal Z. The principal transformations implemented in this synthesis include two highly diastereoselective radical cyclization reactions: a Sm^(II)-mediated reductive cascade cyclization, which forms two rings and establishes four new stereocenters in a single step, and a Ti^(III)-mediated reductive epoxide-acrylate coupling that yields a functionalized spirolactone product, which forms a core bicycle of maoecrystal Z.

The preparation of two additional ent-kauranoid natural products, (–)-trichorabdal A and (–)-longikaurin E, is also described from a derivative of this key spirolactone. These syntheses are additionally enabled by the palladium-mediated oxidative cyclization reaction of a silyl ketene acetal precursor that is used to install the bridgehead all-carbon quaternary stereocenter and bicyclo[3.2.1]octane present in each natural product. These studies have established a synthetic relationship among three architecturally distinct ent-kaurane diterpenoids and have forged a path for the preparation of interesting unnatural ent-kauranoid structural analogs for more thorough biological study.

Computationally Guided Monomerization of Red Fluorescent Proteins of the Class Anthozoa

Red fluorescent proteins (RFPs) have attracted significant engineering focus because of the promise of near infrared fluorescent proteins, whose light penetrates biological tissue, and which would allow imaging inside of vertebrate animals. The RFP landscape, which numbers ~200 members, is mostly populated by engineered variants of four native RFPs, leaving the vast majority of native RFP biodiversity untouched. This is largely due to the fact that native RFPs are obligate tetramers, limiting their usefulness as fusion proteins. Monomerization has imposed critical costs on these evolved tetramers, however, as it has invariably led to loss of brightness, and often to many other adverse effects on the fluorescent properties of the derived monomeric variants. Here we have attempted to understand why monomerization has taken such a large toll on Anthozoa class RFPs, and to outline a clear strategy for their monomerization. We begin with a structural study of the far-red fluorescence of AQ143, one of the furthest red emitting RFPs. We then try to separate the problem of stable and bright fluorescence from the design of a soluble monomeric β-barrel surface by engineering a hybrid protein (DsRmCh) with an oligomeric parent that had been previously monomerized, DsRed, and a pre-stabilized monomeric core from mCherry. This allows us to use computational design to successfully design a stable, soluble, fluorescent monomer. Next we took HcRed, which is a previously unmonomerized RFP that has far-red fluorescence (λemission = 633 nm) and attempted to monomerize it making use of lessons learned from DsRmCh. We engineered two monomeric proteins by pre-stabilizing HcRed’s core, then monomerizing in stages, making use of computational design and directed evolution techniques such as error-prone mutagenesis and DNA shuffling. We call these proteins mGinger0.1 (λem = 637 nm / Φ = 0.02) and mGinger0.2 (λem = 631 nm Φ = 0.04). They are the furthest red first generation monomeric RFPs ever developed, are significantly thermostabilized, and add diversity to a small field of far-red monomeric FPs. We anticipate that the techniques we describe will be facilitate future RFP monomerization, and that further core optimization of the mGingers may allow significant improvements in brightness.