Health and wealth in the Roman Empire

Ancient Rome was the largest and most populous empire of its time, and the largest pre-industrial state in European history. Recent though not universally accepted research suggests that at least for the most populous central periods of its history standard of living was also rather higher than before or after. To trace whether this is also reflected in Roman biological standard of living, we present the first large and more or less comprehensive dataset, based on skeletal data for some 10,000 individuals, covering all periods of Roman history, and all regions (even if inevitably unequally). We discuss both the methodologies that we developed and the historical results. Instead of reconstructing heights from the long bones assuming fixed body proportions or from one individual long bone, we apply exploratory factor analysis and calculate factor scores for 50-year periods. Our measure of the biological standard of living declined during the last two centuries B.C. and started to improve again, slowly at first, from the second century A.D. It correlated negatively with population, but also with other aspects of standard of living such as wages or diets

Mutagenesis-Based Characterization and Improvement of a Novel Inclusion Body Tag

Whereas, bacterial inclusion bodies (IBs) for long were regarded as undesirable aggregates emerging during recombinant protein production, they currently receive attention as promising nanoparticulate biomaterials with diverse applications in biotechnology and biomedicine. We previously identified ssTorA, a signal sequence that normally directs protein export via the Tat pathway in , as a tag that induces the accumulation of fused proteins into IBs under overexpression conditions. Here, we used targeted mutagenesis to identify features and motifs being either critical or dispensable for IB formation. We found that IB formation is neither related to the function of ssTorA as a Tat-signal sequence nor is it a general feature of this family of signal sequences. IB formation was inhibited by co-overexpression of ssTorA binding chaperones TorD and DnaK and by amino acid substitutions that affect the propensity of ssTorA to form an α-helix. Systematic deletion experiments identified a minimal region of ssTorA required for IB formation in the center of the signal sequence. Unbiased genetic screening of a library of randomly mutagenized ssTorA sequences for reduced aggregation properties allowed us to pinpoint residues that are critical to sustain insoluble expression. Together, the data point to possible mechanisms for the aggregation of ssTorA fusions. Additionally, they led to the design of a tag with superior IB-formation properties compared to the original ssTorA sequence

A structurally informed autotransporter platform for efficient heterologous protein secretion and display.

<p>Abstract</p> <p>Background</p> <p>The self-sufficient autotransporter (AT) pathway, ubiquitous in Gram-negative bacteria, combines a relatively simple protein secretion mechanism with a high transport capacity. ATs consist of a secreted passenger domain and a β-domain that facilitates transfer of the passenger across the cell-envelope. They have a great potential for the extracellular expression of recombinant proteins but their exploitation has suffered from the limited structural knowledge of carrier ATs. Capitalizing on its crystal structure, we have engineered the <it>Escherichia coli</it> AT Hemoglobin protease (Hbp) into a platform for the secretion and surface display of heterologous proteins, using the <it>Mycobacterium tuberculosis</it> vaccine target ESAT6 as a model protein.</p> <p>Results</p> <p>Based on the Hbp crystal structure, five passenger side domains were selected and one by one replaced by ESAT6, whereas a β-helical core structure (β-stem) was left intact. The resulting Hbp-ESAT6 chimeras were efficiently and stably secreted into the culture medium of <it>E. coli</it>. On the other hand, Hbp-ESAT6 fusions containing a truncated β-stem appeared unstable after translocation, demonstrating the importance of an intact β-stem. By interrupting the cleavage site between passenger and β-domain, Hbp-ESAT6 display variants were constructed that remain cell associated and facilitate efficient surface exposure of ESAT6 as judged by proteinase K accessibility and whole cell immuno-EM analysis. Upon replacement of the passenger side domain of an alternative AT, EspC, ESAT6 was also efficiently secreted, showing the approach is more generally applicable to ATs. Furthermore, Hbp-ESAT6 was efficiently displayed in an attenuated <it>Salmonella typhimurium</it> strain upon chromosomal integration of a single encoding gene copy, demonstrating the potential of the Hbp platform for live vaccine development.</p> <p>Conclusions</p> <p>We developed the first structurally informed AT platform for efficient secretion and surface display of heterologous proteins. The platform has potential with regard to the development of recombinant live vaccines and may be useful for other biotechnological applications that require high-level secretion or display of recombinant proteins by bacteria.</p

Limited tolerance towards folded elements during secretion of the autotransporter Hbp

Many virulence factors secreted by pathogenic Gram-negative bacteria belong to the autotransporter (AT) family. ATs consist of a passenger domain, which is the actual secreted moiety, and a β-domain that facilitates the transfer of the passenger domain across the outer membrane. Here, we analysed folding and translocation of the AT passenger, using Escherichia coli haemoglobin protease (Hbp) as a model protein. Dual cysteine mutagenesis, instigated by the unique crystal structure of the Hbp passenger, resulted in intramolecular disulphide bond formation dependent on the periplasmic enzyme DsbA. A small loop tied off by a disulphide bond did not interfere with secretion of Hbp. In contrast, a bond between different domains of the Hbp passenger completely blocked secretion resulting in degradation by the periplasmic protease DegP. In the absence of DegP, a translocation intermediate accumulated in the outer membrane. A similar jammed intermediate was formed upon insertion of a calmodulin folding moiety into Hbp. The data suggest that Hbp can fold in the periplasm but must retain a certain degree of flexibility and/or modest width to allow translocation across the outer membrane

Four lectures

For centuries after the Middle Ages classical antiquity was a civilization to envy and admire. Not surprisingly this faded with the Industrial Revolution, when modern society began to make a clear break with the past. That loss of admiration is reflected in twentieth century histories of antiquity, where the otherness and underdevelopment of antiquity were now increasingly emphasized. Research focussed on potential social and cultural explanations of ancient economic failure, and largely avoi..

Review: J. Rasmus Brandt, Erika Hagelberg, Gro Bjornstad and Sven Ahrens (eds.), LIfe and death in Asia Minor in Hellenistic, Roman and Byzantine times: Studies in Arcaheology and Bioarchaeology. Studies in Funerary Archaeology 10. Oxford: Oxbow 2017 ISBN 978-1-78570-359-1

In one of the most important works of historical scholarship of the last century, Peter Laslett gave a chilling sketch of The World We Have Lost. The book built on the recent advances in historical demography and family reconstitution to describe a preindustrial society where the Grim Reaper was omnipotent, with average life expectancies at birth of 20-35 years, high infant mortality, cruelly interrupted marriages, and many orphaned children. It was a fitting antidote to romantic views of a past where everything was somehow comfortably better. For the ancient world, such family reconstitutions were impossible for lack of suitable sources