Project description:Many ancient parchments are defaced by red or purple maculae associated with localized destruction of collagen fibres. Although the main characteristics of this damage were present in most of the manuscripts analysed by many authors, no common microbial or fungal denominator has been found so far, and little or no correspondence between the microbial or fungal species isolated from materials could be addressed. In this study, culture-independent molecular methods and scanning electron microscopy (SEM) were used to identify fungal and bacterial communities on parchments affected by the purple stains. Protocols for c extraction and nucleic-acid-based strategies were selected for assays examining the community structure of fungi and bacteria on biodeteriorated parchment. Both SEM and molecular analysis detected the presence of bacterial and fungal cells in the damaged areas. Halophilic, halotolerant proteolytic bacterial species were selected by the saline environment provided by the parchment samples. As common microbial denominators, members of the Actinobacteria, mainly Saccharopolyspora spp. and species of Aspergillus, were detected in all investigated cases. It is proposed that a relationship exists between the phenomenon of purple spots on ancient parchments and that of the 'red heat' phenomenon, known to be present in some products manufactured with marine salt.
Project description:D-galactose orally intake ameliorate DNCB-induced atopic dermatitis by modulating microbiota composition and quorum sensing. The increased abundance of bacteroidetes and decreased abundance of firmicutes was confirmed. By D-galactose treatment, Bacteroides population was increased and prevotella, ruminococcus was decreased which is related to atopic dermatitis.
Project description:In this study, an inkjet bioprinting-based high-throughput screening (HTS) system was designed and applied for the first time to a catecholpyrimidine-based small molecule library to find hit compounds that inhibit c-Jun NH2-terminal kinase1 (JNK1). JNK1 kinase, inactivated MAPKAPK2, and specific fluorescent peptides along with bioink were printed on parchment paper under optimized printing conditions that did not allow rapid evaporation of printed media based on Triton-X and glycerol. Subsequently, different small compounds were printed and tested against JNK1 kinase to evaluate their degree of phosphorylation inhibition. After printing and incubation, fluorescence intensities from the phosphorylated/nonphosphorylated peptide were acquired for the % phosphorylation analysis. The IM50 (inhibitory mole 50) value was determined as 1.55 × 10-15 mol for the hit compound, 22. Thus, this work demonstrated that inkjet bioprinting-based HTS can potentially be adopted for the drug discovery process using small molecule libraries, and cost-effective HTS can be expected to be established based on its low nano- to picoliter printing volume.
Project description:Ion beam analysis plays an important role in cultural heritage (CH) studies as it offers a combination of simultaneous and complementary analytical techniques (PIXE/PIGE/RBS) and spatially resolved mapping functions. Despite being considered non-destructive, the potential risk of beam-induced modifications during analysis is increasingly discussed. This work focuses on the impact of proton beams on parchment, present in our CH in form of unique historical manuscripts. Parchment is one of the organic, protein-based CH materials believed to be the most susceptible to radiation-induced changes. Various modification patterns, observed on parchment cross-sections by optical and electron microscopy are reported: discoloration (yellowing), formation of cavities and denaturation of collagen fibers. Considerable modifications were detected up to 100 µm deep into the sample for beam fluences of 4 µC/cm2 and higher. The presence of ultramarine paint on the parchment surface appears to increase the harmful effects of proton radiation. Based on our results, a maximum radiation dose of 0.5 µC/cm2 can be considered as 'safe boundary' for 2.3 MeV PIXE analysis of parchment under the applied conditions.
Project description:Chromosomes are composed of enormously long DNA molecules which must be distributed correctly as the cells grow and divide. In Escherichia coli the new DNA behind the replication forks is specifically bound by the SeqA protein. SeqA binds to GATC sequences which are methylated on the A of the old strand but not on the new strand. Binding lasts for a period of time until Dam methyltransferase methylates the new strand. It is therefore believed that a region of hemi-methylated DNA covered by SeqA follows the replication fork. We show that this is indeed the case by using global ChIP on Chip analysis and a newly developed method for methylation analysis. A comparison of rapid and slow growth conditions showed that in cells with multiple replication forks per chromosome, the old forks bind little SeqA. Analysis of strains with strong SeqA binding sites at different chromosomal loci supported this finding. The results indicate that a re-organization of the chromosome occurs at a timepoint when new forks have travelled about 20% and old forks about 75% of the way to the terminus. This timepoint coincides with the end of origin sequestration. It is so far not known what brings about the end of origin sequestration. Here we suggest that a reorganization event occurs resulting in both origin desequestration and loss of old replication forks from the SeqA structures.
Project description:Parchment is an ancient writing support formed from dehaired animal skins. Its manufacture comprises a series of liming and scraping steps before being stretched and dried under tension. Historical parchment represents a valuable source of cultural heritage which, until now, has limited investigations to noninvasive analyses to infer ink composition, degradation, or physical changes over time. We highlight the prospect of the molecular and isotope compositions of animal lipids from parchment as an untapped record of its production and the animal's diet and environment. We report a minimally invasive, total lipid extraction aided by a vacuum for historical parchments. The quantitative and qualitative compositions of lipid extracts obtained using this method are compared with those obtained using invasive sampling for nine sacrificial membranes dated 1765-1825 CE. This extraction method is then applied to membranes from the Chancery Parliament Rolls (1814-1820 CE) held by The National Archives, UK to obtain lipids and derive taxonomic and dietary information using their stable carbon isotope compositions. This novel vacuum-aided extraction allows, for the first time, animal lipids to be obtained from parchment minimally invasively, paving the way for dietary and paleoclimate studies using this well-dated and common material.
Project description:Parchment represents an invaluable cultural reservoir. Retrieving an additional layer of information from these abundant, dated livestock-skins via the use of ancient DNA (aDNA) sequencing has been mooted by a number of researchers. However, prior PCR-based work has indicated that this may be challenged by cross-individual and cross-species contamination, perhaps from the bulk parchment preparation process. Here we apply next generation sequencing to two parchments of seventeenth and eighteenth century northern English provenance. Following alignment to the published sheep, goat, cow and human genomes, it is clear that the only genome displaying substantial unique homology is sheep and this species identification is confirmed by collagen peptide mass spectrometry. Only 4% of sequence reads align preferentially to a different species indicating low contamination across species. Moreover, mitochondrial DNA sequences suggest an upper bound of contamination at 5%. Over 45% of reads aligned to the sheep genome, and even this limited sequencing exercise yield 9 and 7% of each sampled sheep genome post filtering, allowing the mapping of genetic affinity to modern British sheep breeds. We conclude that parchment represents an excellent substrate for genomic analyses of historical livestock.