Development and application of analytical methods for the identification of dyes on historical textiles

The analytical investigation of several historically important natural yellow and red dye sources is presented, extending previous work on the chemical characterisation of the acid hydrolysed extracts from dyed yarns. The PDA HPLC studies of weld (Reseda luteola L.) dyed yarn extracts found that neither the substrate nor additional steps in the dyeing process significantly altered the relative ratio of the main flavonoid components. Furthermore, their relative photo-degradation rates on the dyed yarns were similar, presenting few problems in the identification of the biological source. In contrast, the relative ratio of the main flavonoid components in dyer’s greenweed (Genista tinctoria L.) dyed yarn extracts was highly dependent on the dyeing process. The PDA HPLC and HPLC ESI MSn investigation of the acid hydrolysed extracts of yarn dyed with the Serratula species, sawwort (Serratula tinctoria L.) and Serratula coronata L., identified their characteristic dyeing components to be luteolin, apigenin, quercetin, kaempferol and 3-O-methylquercetin. Similar studies performed on young fustic (Cotinus coggygria Scop.) dyed yarn extracts identified the main colouring components to be fisetin and sulfuretin. Due to the relatively high photo-degradation rates of many of the characteristic components in both sawwort and young fustic, the identification of historical yarns dyed with these sources was problematic. The dye components characterisic for Mexican cochineal (Dactylopius coccus Costa) were all found to be structurally related anthraquinones, with similar photo-degradation rates on the dyed yarns. Analysis of the principal dye components from brazilwood (Caesalpinia sappan L.) and logwood (Haematoxylon campechianum L.) using negative ion ESI MSn enabled the identification of new, structure dependent, mass spectrometric breakdown pathways. The mechanisms were supported by deuterium labelling experiments and the results were shown to be useful for the structural determination of previously unidentified components. An elimination product observed in the PDA HPLC analysis of hematein dyed yarn extracts was structurally characterised using NMR techniques. A similar component, derived from brazilein, was also observed in brazilwood dyed yarn extracts. A novel approach towards mordant identification using inductively coupled plasma techniques was investigated. ICP MS analysis was successful in identifying the metallic ions present in the acid hydrolysed extracts from both reference and historical yarns. However, interpretation of the results from historical samples was challenging due to the observation of large amounts of both aluminium and iron in the sample extracts. Within the Monitoring of Damage to Historic Tapestries (MODHT) project, yellow and green yarns from a selection of 15th – 17th century tapestries were investigated. Analysis of the yarn extracts identified weld and dyer’s greenweed as the most prevalent yellow dye sources, with young fustic frequency identified in the metal thread core extracts

The chemical characterisation by HPLC–PDA and HPLC–ESI–MS of unaged and aged fibre samples dyed with sawwort (Serratula tinctoria L.)

The acid-hydrolysed extracts of freshly dyed reference fibres of sawwort harvested from several different geographical locations were characterised by the use of high-performance liquid chromatography with photodiode array detection (HPLC–PDA) and coupled with electrospray ionisation mass spectrometric analysis (HPLC–ESI–MS). A related species, Serratula coronata L. was also characterised. The flavonols quercetin, 3-O-methylquercetin and kaempferol, and the flavones luteolin and apigenin were observed in all samples. Accelerated ageing studies confirmed the sensitivity of the flavonol components to photo-oxidative degradation. The poor lightfastness and small relative proportion of these flavonol components found in the extracts of freshly dyed sawwort limits their use as sawwort ‘markers’ in historical samples

Phylogeography and genetic diversity of a widespread Old World butterfly, Lampides boeticus (Lepidoptera: Lycaenidae)

<p>Abstract</p> <p>Background</p> <p>Evolutionary genetics provides a rich theoretical framework for empirical studies of phylogeography. Investigations of intraspecific genetic variation can uncover new putative species while allowing inference into the evolutionary origin and history of extant populations. With a distribution on four continents ranging throughout most of the Old World, <it>Lampides boeticus </it>(Lepidoptera: Lycaenidae) is one of the most widely distributed species of butterfly. It is placed in a monotypic genus with no commonly accepted subspecies. Here, we investigate the demographic history and taxonomic status of this widespread species, and screen for the presence or absence of the bacterial endosymbiont <it>Wolbachia</it>.</p> <p>Results</p> <p>We performed phylogenetic, population genetic, and phylogeographic analyses using 1799 bp of mitochondrial sequence data from 57 specimens collected throughout the species' range. Most of the samples (>90%) were nearly genetically identical, with uncorrected pairwise sequence differences of 0 – 0.5% across geographic distances > 9,000 km. However, five samples from central Thailand, Madagascar, northern Australia and the Moluccas formed two divergent clades differing from the majority of samples by uncorrected pairwise distances ranging from 1.79 – 2.21%. Phylogenetic analyses suggest that <it>L. boeticus </it>is almost certainly monophyletic, with all sampled genes coalescing well after the divergence from three closely related taxa included for outgroup comparisons. Analyses of molecular diversity indicate that most <it>L. boeticus </it>individuals in extant populations are descended from one or two relatively recent population bottlenecks.</p> <p>Conclusion</p> <p>The combined analyses suggest a scenario in which the most recent common ancestor of <it>L. boeticus </it>and its sister taxon lived in the African region approximately 7 Mya; extant lineages of <it>L. boeticus </it>began spreading throughout the Old World at least 1.5 Mya. More recently, expansion after population bottlenecks approximately 1.4 Mya seem to have displaced most of the ancestral polymorphism throughout its range, though at least two early-branching lineages still persist. One of these lineages, in northern Australia and the Moluccas, may have experienced accelerated differentiation due to infection with the bacterial endosymbiont <it>Wolbachia</it>, which affects reproduction. Examination of a haplotype network suggests that Australia has been colonized by the species several times. While there is little evidence for the existence of morphologically cryptic species, these results suggest a complex history affected by repeated dispersal events.</p

Historical mystery solved: A multi-analytical approach to the identification of a key marker for the historical use of brazilwood (Caesalpinia spp.) in paintings and textiles

Brazilwood-derived pigments and dyes are found in many historical objects, from European paintings to North American First Nations textiles.</p

Phosphoproteomic screening identifies Rab GTPases as novel downstream targets of PINK1

International audienceMutations in the PTEN-induced kinase 1 (PINK1) are causative of autosomal recessive Parkinson's disease (PD). We have previously reported that PINK1 is activated by mitochondrial depolarisation and phosphorylates serine 65 (Ser 65) of the ubiquitin ligase Parkin and ubiquitin to stimulate Parkin E3 ligase activity. Here, we have employed quantitative phosphoproteomics to search for novel PINK1-dependent phosphorylation targets in HEK (human embry-onic kidney) 293 cells stimulated by mitochondrial depolarisation. This led to the identification of 14,213 phosphosites from 4,499 gene products. Whilst most phosphosites were unaffected, we strikingly observed three members of a sub-family of Rab GTPases namely Rab8A, 8B and 13 that are all phosphorylated at the highly conserved residue of serine 111 (Ser 111) in response to PINK1 activation. Using phospho-specific antibodies raised against Ser 111 of each of the Rabs, we demonstrate that Rab Ser 111 phosphoryla-tion occurs specifically in response to PINK1 activation and is abolished in HeLa PINK1 knockout cells and mutant PINK1 PD patient-derived fibroblasts stimulated by mitochondrial depolari-sation. We provide evidence that Rab8A GTPase Ser 111 phosphory-lation is not directly regulated by PINK1 in vitro and demonstrate in cells the time course of Ser 111 phosphorylation of Rab8A, 8B and 13 is markedly delayed compared to phosphorylation of Parkin at Ser 65. We further show mechanistically that phosphorylation at Ser 111 significantly impairs Rab8A activation by its cognate guanine nucleotide exchange factor (GEF), Rabin8 (by using the Ser111Glu phosphorylation mimic). These findings provide the first evidence that PINK1 is able to regulate the phosphorylation of Rab GTPases and indicate that monitoring phosphorylation of Rab8A/ 8B/13 at Ser 111 may represent novel biomarkers of PINK1 activity in vivo. Our findings also suggest that disruption of Rab GTPase-mediated signalling may represent a major mechanism in the neurodegenerative cascade of Parkinson's disease

Historical textile dyeing with Genista tinctoria L.:a comprehensive study by UPLC-MS/MS analysis

Polyphenolic components from Genista species have been well characterised because of their potential as antioxidants and as therapeutic leads; however, the identification of dyer's greenweed (Genista tinctoria L.) in historical textiles has been the subject of only limited studies. This paper presents a comprehensive UPLC-PDA MS/MS study of reference and historical yarns dyed with this species. Several so far unreported dye components that could assist with the identification of this dye source, were characterised by MS/MS. Furthermore, the effect of photo-degradation and textile preparation techniques (such as over-dyeing) on the dye fingerprint was investigated and the results correlated with those obtained from historical samples from the Burrell and Bodleian collections, UK

A global phylogeny of butterflies reveals their evolutionary history, ancestral hosts and biogeographic origins

Butterflies are a diverse and charismatic insect group that are thought to have evolved with plants and dispersed throughout the world in response to key geological events. However, these hypotheses have not been extensively tested because a comprehensive phylogenetic framework and datasets for butterfly larval hosts and global distributions are lacking. We sequenced 391 genes from nearly 2,300 butterfly species, sampled from 90 countries and 28 specimen collections, to reconstruct a new phylogenomic tree of butterflies representing 92% of all genera. Our phylogeny has strong support for nearly all nodes and demonstrates that at least 36 butterfly tribes require reclassification. Divergence time analyses imply an origin similar to 100 million years ago for butterflies and indicate that all but one family were present before the K/Pg extinction event. We aggregated larval host datasets and global distribution records and found that butterflies are likely to have first fed on Fabaceae and originated in what is now the Americas. Soon after the Cretaceous Thermal Maximum, butterflies crossed Beringia and diversified in the Palaeotropics. Our results also reveal that most butterfly species are specialists that feed on only one larval host plant family. However, generalist butterflies that consume two or more plant families usually feed on closely related plants

Metformin reduces liver glucose production by inhibition of fructose-1-6-bisphosphatase.

Metformin is a first-line drug for the treatment of individuals with type 2 diabetes, yet its precise mechanism of action remains unclear. Metformin exerts its antihyperglycemic action primarily through lowering hepatic glucose production (HGP). This suppression is thought to be mediated through inhibition of mitochondrial respiratory complex I, and thus elevation of 5'-adenosine monophosphate (AMP) levels and the activation of AMP-activated protein kinase (AMPK), though this proposition has been challenged given results in mice lacking hepatic AMPK. Here we report that the AMP-inhibited enzyme fructose-1,6-bisphosphatase-1 (FBP1), a rate-controlling enzyme in gluconeogenesis, functions as a major contributor to the therapeutic action of metformin. We identified a point mutation in FBP1 that renders it insensitive to AMP while sparing regulation by fructose-2,6-bisphosphate (F-2,6-P2), and knock-in (KI) of this mutant in mice significantly reduces their response to metformin treatment. We observe this during a metformin tolerance test and in a metformin-euglycemic clamp that we have developed. The antihyperglycemic effect of metformin in high-fat diet-fed diabetic FBP1-KI mice was also significantly blunted compared to wild-type controls. Collectively, we show a new mechanism of action for metformin and provide further evidence that molecular targeting of FBP1 can have antihyperglycemic effects

Dimethyl fumarate in patients admitted to hospital with COVID-19 (RECOVERY): a randomised, controlled, open-label, platform trial

Dimethyl fumarate (DMF) inhibits inflammasome-mediated inflammation and has been proposed as a treatment for patients hospitalised with COVID-19. This randomised, controlled, open-label platform trial (Randomised Evaluation of COVID-19 Therapy [RECOVERY]), is assessing multiple treatments in patients hospitalised for COVID-19 (NCT04381936, ISRCTN50189673). In this assessment of DMF performed at 27 UK hospitals, adults were randomly allocated (1:1) to either usual standard of care alone or usual standard of care plus DMF. The primary outcome was clinical status on day 5 measured on a seven-point ordinal scale. Secondary outcomes were time to sustained improvement in clinical status, time to discharge, day 5 peripheral blood oxygenation, day 5 C-reactive protein, and improvement in day 10 clinical status. Between 2 March 2021 and 18 November 2021, 713 patients were enroled in the DMF evaluation, of whom 356 were randomly allocated to receive usual care plus DMF, and 357 to usual care alone. 95% of patients received corticosteroids as part of routine care. There was no evidence of a beneficial effect of DMF on clinical status at day 5 (common odds ratio of unfavourable outcome 1.12; 95% CI 0.86-1.47; p = 0.40). There was no significant effect of DMF on any secondary outcome