Whole-genome sequencing (WGS) permits comprehensive cancer genome analyses, revealing mutational signatures, imprints of DNA damage and repair processes that have arisen in each patient's cancer. We ...performed mutational signature analyses on 12,222 WGS tumor-normal matched pairs, from patients recruited via the UK National Health Service. We contrasted our results to two independent cancer WGS datasets, the International Cancer Genome Consortium (ICGC) and Hartwig Foundation, involving 18,640 WGS cancers in total. Our analyses add 40 single and 18 double substitution signatures to the current mutational signature tally. Critically, we show for each organ, that cancers have a limited number of 'common' signatures and a long tail of 'rare' signatures. We provide a practical solution for utilizing this concept of common versus rare signatures in future analyses.
Gilles de la Tourette syndrome is a hereditary, neuropsychiatric movement disorder with reported abnormalities in the neurotransmission of dopamine and γ-aminobutyric acid (GABA). Spatially focalized ...alterations in excitatory, inhibitory and modulatory neurochemical ratios within specific functional subdivisions of the basal ganglia, may lead to the expression of diverse motor and non-motor features as manifested in Gilles de la Tourette syndrome. Current treatment strategies are often unsatisfactory thus provoking the need for further elucidation of the underlying pathophysiology. In view of (i) the close spatio-temporal synergy exhibited between excitatory, inhibitory and modulatory neurotransmitter systems; (ii) the crucial role played by glutamate (Glu) in tonic/phasic dopaminergic signalling; and (iii) the interdependent metabolic relationship exhibited between Glu and GABA via glutamine (Gln); we postulated that glutamatergic signalling is related to the pathophysiology of Gilles de la Tourette syndrome. As such, we examined the neurochemical profile of three cortico-striato-thalamo-cortical regions in 37 well-characterized, drug-free adult patients and 36 age/gender-matched healthy control subjects via magnetic resonance spectroscopy at 3 T. To interrogate the influence of treatment on metabolite concentrations, spectral data were acquired from 15 patients undergoing a 4-week treatment with aripiprazole. Test-retest reliability measurements in 23 controls indicated high repeatability of voxel localization and metabolite quantitation. We report significant reductions in striatal concentrations of Gln, Glu + Gln (Glx) and the Gln:Glu ratio, and thalamic concentrations of Glx in Gilles de la Tourette syndrome in comparison to controls. ON-treatment patients exhibited no significant metabolite differences when compared to controls but significant increases in striatal Glu and Glx, and trends for increases in striatal Gln and thalamic Glx compared to baseline measurements. Multiple regression analysis revealed a significant negative correlation between (i) striatal Gln and actual tic severity; and (ii) thalamic Glu and premonitory urges. Our results indicate that patients with Gilles de la Tourette syndrome exhibit an abnormality in the flux of metabolites in the GABA-Glu-Gln cycle, thus implying perturbations in astrocytic-neuronal coupling systems that maintain the subtle balance between excitatory and inhibitory neurotransmission within subcortical nuclei.
The impact of psychosocial stress on a variety of negative health outcomes is well documented, with current research efforts directed at possible mechanisms. Here, we focused on a potential mechanism ...involving differential expression of mRNA and microRNA in response to acute psychosocial stress. We utilized a validated behavioral paradigm, the Trier Social Stress Test (TSST), to induce acute psychosocial stress in a cohort of volunteers. Stress reactivity was assessed repeatedly during the TSST using saliva samples that were analyzed for levels of cortisol. Peripheral blood mononuclear cells were extracted from blood drawn at baseline and at two time points following the stress paradigm. Total RNA was extracted, and mRNA and microRNA microarrays were utilized to assess within-subject changes in gene expression between baseline and the two post-stressor time points.
For microarray gene expression analysis, we focused on 12 participants who showed a robust cortisol response to the task, as an indicator of robust HPA-axis activation. We discovered a set of mRNAs and miRNAs that exhibited dynamic expression change in response to the TSST in peripheral blood mononuclear cells, further characterizing the link between psychosocial stress and cellular response mechanisms.
Background:
Increased posterior tibial slope (PTS) is a risk factor for knee pathology. Accurate measurement of PTS is predicated on a quality lateral knee radiograph; however, little is known about ...how the quality of the radiograph affects the measured PTS.
Purposes:
To (1) describe a method for measuring malalignment on lateral knee radiographs, (2) assess the effects of malpositioning of the knee on radiographic measures of malalignment, and (3) determine any correlations between malalignment and the measured PTS.
Study Design:
Descriptive laboratory study.
Methods:
Using a setup similar to that of a standard radiology suite, 25 sets of radiographs were taken using 5 sawbone models. Each set included a true lateral view and separate malpositioned radiographs at 5°, 10°, and 15° of adduction, abduction, internal rotation, and external rotation. Malalignment for each radiograph was quantified as the anterior-posterior distance (APD) and proximal-distal distance (PDD) between femoral condyles. The medial PTS was measured in duplicate, and the interrater reliability was calculated.
Results:
The interrater reliability was excellent, with intraclass correlation coefficients of 0.92, 0.91, and 0.96 for the APD, PDD, and PTS, respectively. Malrotation significantly affected the APD (P < .001), with a mean change of 5.6 mm per 5°. Malpositioning in abduction/adduction significantly affected the PDD (P < .001), with a mean change of 5.1 mm per 5°. There was no significant impact of rotation or APD on the PTS. Abduction/adduction did affect the PTS (P < .001) above a threshold of 5° of malpositioning. The PTS decreased as the PDD increased, moving from adduction to abduction (R
2 = 0.5687).
Conclusion:
The measured PTS was more sensitive to malpositioning by abduction/adduction than by malrotation. Malrotation affected the APD, while abduction/adduction affected the PDD. Thus, the accuracy of the measured PTS was compromised more by poorly aligned distal femoral condyles than it was by poorly aligned posterior femoral condyles.
Clinical Relevance:
To minimize the effects of malpositioning, we recommend utilizing radiographs with a |PDD| of <5 mm and an |APD| of <15 mm when measuring the PTS.
Abstract
CD40 ligand is a member of the TNF superfamily (TNF-SF) and a key regulator of the immune system. Its cognate receptor CD40 is expressed on antigen-presenting cells and on many tumor types, ...and has emerged as an attractive target for immunological cancer treatment. We developed HERA-CD40L, a construct composed of a trivalent single-chain CD40L-receptor-binding-domain (scCD40L-RBD) linked to a silenced human IgG1-Fc-domain thereby generating a hexavalent molecule. We showed previously that HERA-CD40L mimics the natural ligand, thereby inducing potent agonistic activity which is independent of FcγR mediated crosslinking and superior to anti-CD40 benchmark antibodies. For combinatorial cancer immunotherapy we have created bispecific molecules by combining the HERA-CD40L scaffold with antibody derived domains targeting different classes of tumor associated antigens. These bispecific fusion proteins combine the potent co-stimulatory CD40-agonist with either direct tumor-cell targeting and/or additional immunomodulatory activities in the tumor microenvironment. To evaluate the different fusion protein formats, the tumor associated antigens CEA, PD-L1 and CD95L were chosen as model-targets. In addition to the hexavalent targeted HERA-CD40L, trivalent targeted fusion proteins, employing the scCD40L-RBD as building block, were created as well. All engineering prototypes were produced in CHO-S cells and purified, resulting in highly pure non-aggregating protein lots as demonstrated by SDS-PAGE and HPLC-SEC. Functional binding to their respective targets was shown by ELISA and to proof biological in vitro activity luciferase reporter gene assays were employed. The basic underlying immunological processes have been investigated in vitro. The trivalent CD40L, the trivalent CD40L-bispecifics anti-CD95L-scCD40L-RBD and anti-CEA-scCD40L-RBD as well as anti-CD40, anti-PD-L1 or anti-CD95L antibodies did not activate human monocytes, even if the antibodies were co-administered with trivalent CD40L. In contrast, the hexavalent HERA-CD40L, the hexavalent bifunctional CD40L-constructs and the trivalent anti-PD-L1-scCD40L-RBD-construct induced strong activation/maturation of the monocytes as indicated by CD83, CD86, HLA-DR upregulation which was accompanied by increased chemokine receptor (CD54, CCR7) and PD-L1 expression. When co-cultured with CD3 positive T cells, these pre-activated monocytes led to subsequent activation of CD4 as well as CD8 positive T cells, indicated by increased expression of CD25, CD69 and CD54. This clearly shows that the close proximity of anti-PD-L1 and trivalent CD40L within one molecule makes a huge difference for biological activity. Hence, these novel bispecific constructs are a promising therapeutic approach to promote anti-tumor immune responses.
Citation Format: Matthias Schroeder, Katharina Billian-Frey, Jaromir Sykora, Mauricio Redondo-Mueller, Karl Heinonen, Jamie Frankish, Christian Gieffers, Meinolf Thiemann, Oliver Hill. Generation and characterization of novel bispecific molecules combining single-chain-CD40L with anti-CEA, anti-CD95L or anti-PD-L1 targeting moieties abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1587.
Our skin influences our physical and mental health, and its chemical composition can reflect environmental and disease conditions. Therefore, through sampling the skin metabolome, we can provide a ...promising window into the mechanisms of the body. However, the broad application of skin metabolomics has recently been hampered by a lack of easy and widely applicable sampling methods. Here, we present a novel rapid, simple, and, most importantly, painless and non-invasive sampling technique suitable for clinical studies of fragile or weakened skin. The method is called WET PREP and is simply a lavage of the skin which focuses on capturing the metabolome. We systematically evaluate WET PREPs in comparison with the non-invasive method of choice in skin metabolomics, swab collection, using ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-MS2) on two complementary chromatographic columns (C18 reversed phase and hydrophilic interaction chromatography). We also integrate targeted analyses of key metabolites of skin relevance. Overall, WET PREP provides a strikingly more stable shared metabolome across sampled individuals, while also being able to capture unique individual metabolites with a high consistency in intra-individual reproducibility. With the exception of (phospho-)lipidomic studies, we recommend WET PREPs as the preferred skin metabolome sampling technique due to the quick preparation time, low cost, and gentleness for the patient.
This exploratory case study describes the design and facilitation of a massive open online course (MOOC) for attitudinal change regarding human trafficking. It examines the course from the learners', ...instructor's, and instructional designer's perspectives. Two interviews with the instructor and instructional designer were conducted, and data from a sample of learners via an end-of-course survey (n = 54) and follow-up questionnaire (n = 319) were gathered. Learners' discussion posts and sample assignments were also reviewed. Findings show that the instructor and instructional designer perceived the design and facilitation of the MOOC as highly complex and challenging. Learner feedback was contradictory, possibly due to different expectations and needs within the MOOC. Six instructional design considerations for MOOCs in general and for attitudinal change are discussed, including: (a) MOOCs as a unique platform for attitudinal change, (b) the support needed from platform providers and universities, (c) personal and flexible learning paths, (d) instructional activities for attitudinal dissonance, (e) creating a collaborative community, and (f) MOOC instructor preparation.
Comparative analysis of RNA sequences is the basis for the detailed and accurate predictions of RNA structure and the determination of phylogenetic relationships for organisms that span the entire ...phylogenetic tree. Underlying these accomplishments are very large, well-organized, and processed collections of RNA sequences. This data, starting with the sequences organized into a database management system and aligned to reveal their higher-order structure, and patterns of conservation and variation for organisms that span the phylogenetic tree, has been collected and analyzed. This type of information can be fundamental for and have an influence on the study of phylogenetic relationships, RNA structure, and the melding of these two fields.
We have prepared a large web site that disseminates our comparative sequence and structure models and data. The four major types of comparative information and systems available for the three ribosomal RNAs (5S, 16S, and 23S rRNA), transfer RNA (tRNA), and two of the catalytic intron RNAs (group I and group II) are: (1) Current Comparative Structure Models; (2) Nucleotide Frequency and Conservation Information; (3) Sequence and Structure Data; and (4) Data Access Systems.
This online RNA sequence and structure information, the result of extensive analysis, interpretation, data collection, and computer program and web development, is accessible at our Comparative RNA Web (CRW) Site http://www.rna.icmb.utexas.edu. In the future, more data and information will be added to these existing categories, new categories will be developed, and additional RNAs will be studied and presented at the CRW Site.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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