Correction: Quantifying spatial variability in shell midden formation in the Farasan Islands, Saudi Arabia.

[This corrects the article DOI: 10.1371/journal.pone.0217596.].

Applying laser induced breakdown spectroscopy (LIBS) and elemental imaging on marine shells for archaeological and environmental research.

Using LIBS for the analysis of archaeological and geological marine mollusc shells is a growing research area that relies on customised instrumentation and specific workflows that can accommodate the variety and precision of the required sampling parameters. However, the increased efficiency offered by LIBS, which enables the study of a larger quantity of shell samples for temperature variation, ecological parameters, and human consumption practices, outweighs the initial efforts required to develop customised instrumentation and workflows. In this work, we present detailed specifications and parameters for the development of a LIBS system capable of generating Mg/Ca images on marine shells that directly correlate with seasonal sea temperatures. Our main objective was to develop specifications that enable easy adaptation of LIBS systems to existing laboratories for studying hard-tissue samples. These specifications were used to develop a customised micro-LIBS system and apply it to a real-world example of an archaeological study to better understand its efficiency on the marine mollusc shells and demonstrate its potential for broader applications in interdisciplinary research. In total 101 shell specimens have been analysed within a time frame of approximately 71 h of machine time, producing 234 images (100 µm resolution: 100 images, 30 µm resolution: 134 images). SEM analysis of the irradiated sections of the shell revealed a primary ablated area of 10-15 µm in diameter, while a secondary affected area of the shell's crystal fabric extended to 30-50 µm after repeated shots. Overall, this new customised system reliably and efficiently analysed marine mollusc specimens without major destructive effects, enabling additional analyses for other proxies to be carried out. This study highlights the potential of the LIBS method for interdisciplinary research, encompassing applications in paleoclimatology, marine ecology, and archaeology.

Quantifying spatial variability in shell midden formation in the Farasan Islands, Saudi Arabia.

During the past decade, over 3000 shell middens or shell matrix deposits have been discovered on the Farasan Islands in the southern Red Sea, dating to the period c. 7,360 to 4,700 years ago. Many of the sites are distributed along a palaeoshoreline which is now 2-3 m above present sea level. Others form clusters with some sites on the shoreline and others located inland over distances of c. 30 m to 1 km. We refer to these inland sites as 'post-shore' sites. Following Meehan, who observed a similar spatial separation in shell deposition in her ethnographic study of Anbarra shellgathering in the Northern Territory of Australia, we hypothesise that the shoreline sites are specialised sites for the processing or immediate consumption of shell food, and the post-shore sites are habitation sites used for a variety of activities. We test this proposition through a systematic analysis of 55 radiocarbon dates and measurement of shell quantities from the excavation of 15 shell matrix sites in a variety of locations including shoreline and post-shore sites. Our results demonstrate large differences in rates of shell accumulation between these two types of sites and selective removal of shoreline sites by changes in sea level. We also discuss the wider implications for understanding the differential preservation and visibility of shell-matrix deposits in coastal settings in other parts of the world extending back into the later Pleistocene in association with periods of lowersea level. Our results highlight the importance of taphonomic factors of post-depositional degradation and destruction, rates of shell accumulation, the influence on site location of factors other than shell food supply, and the relative distance of deposits from their nearest palaeoshorelines as key variables in the interpretation of shell quantities. Failure to take these variables into account when investigating shells and shell-matrix deposits in late Pleistocene and early Holocene contexts is likely to compromise interpretations of the role and significance of shell food in human evolutionary and socio-cultural development.

Extending SILAC to proteomics of plant cell lines.

Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) is a widespread method for metabolic labeling of cells and tissues in quantitative proteomics; however, incomplete incorporation of the label has so far restricted its wider use in plants. Here, we argue that differential labeling by two different versions of the labeled amino acids renders SILAC fully applicable to dark-grown plant cell lines. By comparing Arabidopsis thaliana cell cultures labeled with two versions of heavy Lys (Lys-4 and Lys-8), we show that this simple modification of the SILAC protocol enables similar quantitation accuracy, precision, and reproducibility as conventional SILAC in animal cells.