The Effects of Possible Contamination on the Radiocarbon Dating of the Dead Sea Scrolls II: Empirical Methods to Remove Castor Oil and Suggestions for Redating

While kept at the Rockefeller Museum in East Jerusalem, many Dead Sea Scroll fragments were exposed to castor oil by the original team of editors in the course of cleaning the parchments. Castor oil must be regarded as a serious contaminant in relation to radiocarbon dating. If modern castor oil is present and is not removed prior to dating, the 14C dates will be skewed artificially towards modern values. Earlier, it was shown that the standard AAA pretreatment procedure used in the 2 previous studies dating Dead Sea Scroll samples is not capable of removing castor oil from parchment samples. In the present work, we show that it is unlikely that castor oil reacts with the amino acids of the parchment proteins, a finding which leaves open the possibility of devising a cleaning method that can effectively remove castor oil. We then present 3 different pretreatment protocols designed to effectively remove castor oil from parchment samples. These involve 3 different cleaning techniques: extraction with supercritical CO2, ultrasound cleaning, and Soxhlet extraction—each with their own advantages and disadvantages. Our data show that the protocol involving Soxhlet extraction is the best suited for the purpose of decontaminating the Dead Sea Scrolls, and we recommend that this protocol be used in further attempts to 14C date the Dead Sea Scrolls. If such an attempt is decided on by the proper authorities, we propose a list of Scroll texts, which we suggest be redated in order to validate the 14C dates done earlier.

Multi-omics Analyses of Starvation Responses Reveal a Central Role for Lipoprotein Metabolism in Acute Starvation Survival in <i>C. elegans</i>

Starvationcauses comprehensivemetabolicchanges, which are still not fully understood. Here, we used quantitative proteomics and RNA sequencing to examine the temporal starvation responses in wildtype Caenorhabditis elegans and animals lacking the transcription factor HLH-30. Our findings show that starvation alters the abundance of hundreds of proteins and mRNAs in a temporal manner, many of which are involved in central metabolic pathways, including lipoprotein metabolism. We demonstrate that premature death of hlh-30 animals under starvation can be prevented by knockdown of either vit-1 or vit-5, encoding two different lipoproteins. We further showthat the size and number of intestinal lipid droplets under starvation are altered in hlh-30 animals, which can be rescued by knockdown of vit-1. Taken together, this indicates that survival of hlh-30 animals under starvation is closely linked to regulation of intestinal lipid stores. We provide the most detailed poly-omic analysis of starvation responses to date, which serves as a resource for further mechanistic studies of starvation

Increased Protein Kinase A Activity Induces Fibrolamellar Hepatocellular Carcinoma Features Independent of DNAJB1

Fibrolamellar hepatocellular carcinoma (FLC) is a rare liver cancer that is driven by the fusion of DNAJB1 and PRKACA, the catalytic subunit of protein kinase A (PKA). PKA activity is controlled through regulatory proteins that both inhibit catalytic activity and control localization, and an excess of regulatory subunits ensures PRKACA activity is inhibited. Here, we found an increase in the ratio of catalytic to regulatory units in FLC patient tumors driven by DNAJB1::PRKACA using mass spectrometry, biochemistry, and immunofluorescence, with increased nuclear localization of the kinase. Overexpression of DNAJB1::PRKACA, ATP1B1::PRKACA, or PRKACA, but not catalytically inactive kinase, caused similar transcriptomic changes in primary human hepatocytes, recapitulating the changes observed in FLC. Consistently, tumors in patients missing a regulatory subunit or harboring an ATP1B1::PRKACA fusion were indistinguishable from FLC based on the histopathological, transcriptomic, and drug-response profiles. Together, these findings indicate that the DNAJB1 domain of DNAJB1::PRKACA is not required for FLC. Instead, changes in PKA activity and localization determine the FLC phenotype. Significance: Alterations leading to unconstrained protein kinase A signaling, regardless of the presence or absence of PRKACA fusions, drive the phenotypes of fibrolamellar hepatocellular carcinoma, reshaping understanding of the pathogenesis of this rare liver cancer.</p

Structural analysis of dimeric calreticulin using stable isotope labelling, cross-linking, mass spectrometry and modelling

Description of raw-files: FUS04612: Orbitrap Fusion analysis of heat-treated and BS3 cross-linked dimeric calreiticulin where the dimer is a combination of human recombinant 15N-labelled calreticulin expressed in P. pastoris mixed with native human placental calreticulin. FUS04614: Duplicate of FUS04612 QHF1883: Q Exactive HF analysis of heat-treated and BS3 cross-linked dimeric calreiticulin where the dimer is a combination of human recombinant 15N-labelled calreticulin expressed in P. pastoris mixed with native human placental calreticulin. QHF1884: Duplicate of QHF1883 Obvb08125: Orbitrap Velos Pro analysis of heat-treated and BS3 cross-linked dimeric calreiticulin where the dimer is a combination of human recombinant 15N-labelled calreticulin expressed in P. pastoris mixed with native human placental calreticulin. Obvb08126, Obvb08127, Obvb08128: Three replicates of Obvb08125

Insect cuticular proteins

Insect cuticles are composite structural materials with mechanical properties optimal for their biological functions. The bulk properties of cuticles are to a large extent determined by the interactions between the various components, mainly the chitin filament system and the proteins. The various cuticular types show pronounced differences in mechanical properties, and it is suggested that these differences can be related to the properties of the individual proteins and to the degree of secondary stabilization (sclerotization). The amino acid sequences, which have been obtained for insect cuticular proteins either by direct sequencing of purified proteins or by deduction from corresponding DNA-sequences, are listed according to insect order and species. Extensive sequence similarity is observed among several cuticular proteins obtained from different insect orders. Other cuticular proteins are characterized by repeated occurrence of a few small motifs consisting mainly of hydrophobic residues. The latter group of proteins has so far only been reported from stift cuticles. The possible relevance of the various motifs and repeats for protein interaction and the mechanical properties of cuticles is discussed.</p