Genetic mapping studies in the AS/AGU rat

The AS/AGU rat arose by spontaneous mutation from a closed colony of AS rats in the Laboratory of Human Anatomy, The University of Glasgow, in 1990. These rats had locomotor abnormalities and appeared to have a striatal dopamine deficiency syndrome, symptoms and pathology reminiscent of human Parkinson's disease. Test crosses between AS/AGU and AS rats indicated that the mutant locus, agu, was inherited in an autosomal recessive fashion with no evidence of maternal or X-linked involvement. Two reference strains, BN and F344, were used to set up backcrosses with AS/AGU, namely, [AS/AGUxF1(AS/AGUxBN)] and [AS/AGUxF1(AS/AGUxF344)]. Previously, 75 microsatellites markers spanning the rat genome were tested for informativeness between AS/AGU and the strains BN and F344. In this study, 19 other microsatellites were typed on these strains, 9 of which displayed size differences and could therefore be mapped relative to agu. Two candidate chromosomes had been thought to harbour agu, one of which was eliminated in this report. Initially, four markers from the other chromosome had aU shown linkage with agu but none were close enough to commence chromosome walking to the disease locus. This report describes the analysis of pre-existing microsatellites and the search for new ones associated with a gene family believed to be adjacent to agu. A total of twelve markers, associated with this gene family, were analysed for size or sequence variation between AS/AGU and reference strains. None proved informative. It was concluded that members of this gene family had not diverged sufficiently between these strains. Another reference strain, DA, has been shown to display size differences from the AS strain from members of this gene family and other markers believed to be near agu. A backcross program using this reference strain is now underway. To date, another gene-based marker has been shown to be less than 0.5 cM from agu and physical mapping is now underway to track down a gene harbouring this mutation. The eventual discovery of such a gene and its role in normal physiology will prove valuable in the understanding of basal ganglia disorders

From the Mendeleev periodic table to particle physics and back to the periodic table

We briefly describe in this paper the passage from Mendeleev's chemistry (1869) to atomic physics (in the 1900's), nuclear physics (in the 1932's) and particle physics (from 1953 to 2006). We show how the consideration of symmetries, largely used in physics since the end of the 1920's, gave rise to a new format of the periodic table in the 1970's. More specifically, this paper is concerned with the application of the group SO(4,2)xSU(2) to the periodic table of chemical elements. It is shown how the Madelung rule of the atomic shell model can be used for setting up a periodic table that can be further rationalized via the group SO(4,2)xSU(2) and some of its subgroups. Qualitative results are obtained from this nonstandard table.Comment: 15 pages; accepted for publication in Foundations of Chemistry (special issue to commemorate the one hundredth anniversary of the death of Mendeleev who died in 1907); version 2: 16 pages; some sentences added; acknowledgment and references added; misprints correcte

Comparability: manufacturing, characterization and controls, report of a UK Regenerative Medicine Platform Pluripotent Stem Cell Platform Workshop, Trinity Hall, Cambridge, 14–15 September 2015

This paper summarizes the proceedings of a workshop held at Trinity Hall, Cambridge to discuss comparability and includes additional information and references to related information added subsequently to the workshop. Comparability is the need to demonstrate equivalence of product after a process change; a recent publication states that this ‘may be difficult for cell-based medicinal products’. Therefore a well-managed change process is required which needs access to good science and regulatory advice and developers are encouraged to seek help early. The workshop shared current thinking and best practice and allowed the definition of key research questions. The intent of this report is to summarize the key issues and the consensus reached on each of these by the expert delegates

Functional Heterogeneity of Embryonic Stem Cells Revealed through Translational Amplification of an Early Endodermal Transcript

Detection of low-level, lineage-specific transcription aids in the identification of lineage-primed populations of ES cells provides a new framework for pluripotency

Increased Cycling Cell Numbers and Stem Cell Associated Proteins as Potential Biomarkers for High Grade Human Papillomavirus+ve Pre-Neoplastic Cervical Disease

High risk (oncogenic) human papillomavirus (HPV) infection causes cervical cancer. Infections are common but most clear naturally. Persistent infection can progress to cancer. Pre-neoplastic disease (cervical intraepithelial neoplasia/CIN) is classified by histology (CIN1-3) according to severity. Cervical abnormalities are screened for by cytology and/or detection of high risk HPV but both methods are imperfect for prediction of which women need treatment. There is a need to understand the host virus interactions that lead to different disease outcomes and to develop biomarker tests for accurate triage of infected women. As cancer is increasingly presumed to develop from proliferative, tumour initiating, cancer stem cells (CSCs), and as other oncogenic viruses induce stem cell associated gene expression, we evaluated whether presence of mRNA (detected by qRT-PCR) or proteins (detected by flow cytometry and antibody based proteomic microarray) from stem cell associated genes and/or increased cell proliferation (detected by flow cytometry) could be detected in well-characterised, routinely collected cervical samples from high risk HPV+ve women. Both cytology and histology results were available for most samples with moderate to high grade abnormality. We found that stem cell associated proteins including human chorionic gonadotropin, the oncogene TP63 and the transcription factor SOX2 were upregulated in samples from women with CIN3 and that the stem cell related, cell surface, protein podocalyxin was detectable on cells in samples from a subset of women with CIN3. SOX2, TP63 and human gonadotrophin mRNAs were upregulated in high grade disease. Immunohistochemistry showed that SOX2 and TP63 proteins clearly delineated tumour cells in invasive squamous cervical cancer. Samples from women with CIN3 showed increased proliferating cells. We believe that these markers may be of use to develop triage tests for women with high grade cervical abnormality to distinguish those who may progress to cancer from those who may be treated more conservatively

Characterization of ATG8 Family Protein Binding to TRIM23 and TRIM31

Macroautophagy (hereafter called autophagy) is a process where proteins, organelles and intracellular micro-organisms are degraded by the lysosome. The autophagosome engulfs a part of the cytoplasm where the cargo is, and transports it to the endosomal-lysosomal system for degradation. Autophagy can also be selective, where cargos are recognized directly by the autophagy receptors or by other proteins in which target a specific cargo for degradation and can bind to for example ATG8s via a LIR domain. There are many proteins involved in autophagy, and TRIM proteins represent some of them. TRIM proteins are classified in a family of E3 ligases because of their RING domain, even though not all TRIM proteins contain this RING domain. Some TRIM proteins have been shown to be of great importance in the process of autophagy and the selectivity of autophagy by binding to autophagy-related proteins. Recent studies show that TRIM23 and TRIM31 are important in autophagy. ATG8 family proteins have hydrophobic pockets which bind LIR motifs in other proteins. Here, the interaction of TRIM23 and TRIM31 with the ATG8 family proteins is studied, and moreover their interaction with human ATG8 family proteins and if the binding is LIR dependent. GST pulldown, and co-localization assays with confocal fluorescence microscope were methods used for this purpose. Furthermore, the mCherry-EYFP double-tag was used to monitor if TRIM23 and TRIM31 are degraded in the lysosome. The lysosomal inhibitor Bafilomycin A1 (Baf) and the proteosomal inhibitor MG132 were included in the assays performed in HeLa cells to determine the degradation pathway. The results showed that both TRIM23 and TRIM31 bind ATG8s in a LIR-dependent manner. Neither TRIM23 nor TRIM31 were found to be degraded by autophagy using the double-tag assay, even though both were degraded upon starvation. Confocal imaging showed that TRIM31 co-localizes with LC3A, LC3B and GABARAP in HeLa cells