Olaparib-Based Photoaffinity Probes for PARP-1 Detection in Living Cells

The poly-ADP-ribose polymerase (PARP) is a protein from the family of ADP-ribosyltransferases that catalyzes polyadenosine diphosphate ribose (ADPR) formation in order to attract the DNA repair machinery to sites of DNA damage. The inhibition of PARP activity by olaparib can cause cell death, which is of clinical relevance in some tumor types. This demonstrates that quantification of PARP activity in the context of living cells is of great importance. In this work, we present the design, synthesis and biological evaluation of photo-activatable affinity probes inspired by the olaparib molecule that are equipped with a diazirine for covalent attachment upon activation by UV light and a ligation handle for the addition of a reporter group of choice. SDS-PAGE, western blotting and label-free LC-MS/MS quantification analysis show that the probes target the PARP-1 protein and are selectively outcompeted by olaparib; this suggests that they bind in the same enzymatic pocket. Proteomics data are available via ProteomeXchange with identifier PXD018661.     

Glass alteration in 19th century glass photographic plates: Potential role in gelatin delamination

The Smithsonian Institution's National Museum of American History houses the largest remaining collection of inter-positive composite glass plate images from the production of Eadweard Muybridge's Animal Locomotion series. These photographs, which are approximately 150 years old, exhibit a wide range of degradation phenomena and products, including alteration of the glass plates. Compositions of both the image-bearing glass plates and support glass panels were obtained through quantitative scanning electron microscope–energy-dispersive spectrometry. Electron imaging of cross-sectioned samples of the image-bearing glass plates reveals a visible sodium-depleted glass alteration layer. Furthermore, electron imaging shows delamination of the image-containing gelatin layer and captures evidence that suggests that interfacial stresses between the glass alteration layer and the unaltered glass contribute to the delamination of this layer along with the overlaid gelatin.     

Folate Receptor-Mediated Delivery of Cas9 RNP for Enhanced Immune Checkpoint Disruption in Cancer Cells

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system offers great opportunities for the treatment of numerous diseases by precise modification of the genome. The functional unit of the system is represented by Cas9/sgRNA ribonucleoproteins (RNP), which mediate sequence-specific cleavage of DNA. For therapeutic applications, efficient and cell-specific transport into target cells is essential. Here, Cas9 RNP nanocarriers are described, which are based on lipid-modified oligoamino amides and folic acid (FolA)-PEG to realize receptor-mediated uptake and gene editing in cancer cells. In vitro studies confirm strongly enhanced potency of receptor-mediated delivery, and the nanocarriers enable efficient knockout of GFP and two immune checkpoint genes, PD-L1 and PVR, at low nanomolar concentrations. Compared with non-targeted nanoparticles, FolA-modified nanocarriers achieve substantially higher gene editing including dual PD-L1/PVR gene disruption after injection into CT26 tumors in vivo. In the syngeneic mouse model, dual disruption of PD-L1 and PVR leads to CD8+ T cell recruitment and distinct CT26 tumor growth inhibition, clearly superior to the individual knockouts alone. The reported Cas9 RNP nanocarriers represent a versatile platform for potent and receptor-specific gene editing. In addition, the study demonstrates a promising strategy for cancer immunotherapy by permanent and combined immune checkpoint disruption.     

Pilot tone-based prospective correction of respiratory motion for free-breathing myocardial T1 mapping

Objective

To provide respiratory motion correction for free-breathing myocardial T1 mapping using a pilot tone (PT) and a continuous golden-angle radial acquisition.

Materials and methods

During a 45 s prescan the PT is acquired together with a dynamic sagittal image covering multiple respiratory cycles. From these images, the respiratory heart motion in head-feet and anterior–posterior direction is estimated and two linear models are derived between the PT and heart motion. In the following scan through-plane motion is corrected prospectively with slice tracking based on the PT. In-plane motion is corrected for retrospectively. Our method was evaluated on a motion phantom and 11 healthy subjects.

Results

Non-motion corrected measurements using a moving phantom showed T1 errors of 14 ± 4% (p < 0.05) compared to a reference measurement. The proposed motion correction approach reduced this error to 3 ± 4% (p < 0.05). In vivo the respiratory motion led to an overestimation of T1 values by 26 ± 31% compared to breathhold T1 maps, which was successfully corrected to an average difference of 3 ± 2% (p < 0.05) between our free-breathing approach and breathhold data.

Discussion

Our proposed PT-based motion correction approach allows for T1 mapping during free-breathing with the same accuracy as a corresponding breathhold T1 mapping scan.

     

Recycled materials in railroad substructure: an energy perspective

Railway Engineering Science - Given that the current ballasted tracks in Australia may not be able to support faster and significantly heavier freight trains as planned for the future, the imminent...     

An Ontology Based Cyclone Tracks Classification Using SWRL Reasoning and SVM

Abstract: Tropical cyclones (TC) are often associated with severe weather conditions which cause great losses to lives and property. The precise classification of cyclone tracks is significantly important in the field of weather forecasting. In this paper we propose a novel hybrid model that integrates ontology and Support Vector Machine (SVM) to classify the tropical cyclone tracks into four types of classes namely straight, quasi-straight, curving and sinuous based on the track shape. Tropical Cyclone TRacks Ontology (TCTRO) described in this paper is a knowledge base which comprises of classes, objects and data properties that represent the interaction among the TC characteristics. A set of SWRL (Semantic Web Rule Language) rules are directly inserted to the TCTRO ontology for reasoning and inferring new knowledge from ontology. Furthermore, we propose a learning algorithm which utilizes the inferred knowledge for optimizing the feature subset. According to experiments on the IBTrACS dataset, the proposed ontology based SVM classifier achieves an accuracy of 98.3% with reduced classification error rates.     

Engineered Platelet-Derived Growth Factor-Releasing Hydrogels Promote Fetal Membrane Healing In Vivo

Fetoscopic interventions to treat fetal anomalies are currently performed for a variety of conditions. Depending on the procedure, preterm rupture of the fetal membranes (FMs) happens in around 30% of the cases, potentially leading to preterm birth and fetal morbidity and mortality. Here, the capacity of modular transglutaminase crosslinked poly(ethylene glycol) (TG-PEG) hydrogels that release platelet-derived growth factor (PDGF)-BB to promote FM healing is described. In vitro, such growth factor-loaded hydrogels are able to stimulate amniotic cell migration and proliferation. When applied in vivo, these TG-PEG hydrogels tightly seal the FM and uterus defects created by a fetoscope and remain stable for 10 days. The migration of healing-related cells into such hydrogels in the myometrium, endometrium, and FM areas is only possible in soft TG-PEG hydrogels. Importantly, bioengineered hydrogels releasing PDGF-BB promote recruitment of host cells from the myometrium and the endometrium, and to a lesser extent from FM areas. In such hydrogels, the potent proliferation and matrix production of the recruited cells at the site of treatment into the biomaterial initiates a robust early healing response. PDGF-BB-loaded TG-PEG hydrogels hold great promise for the treatment of fetoscopy-induced FM defects and for the prevention of preterm birth.     

High-Resolution Patterning of Organic–Inorganic Photoresins for Tungsten and Tungsten Carbide Microstructures

Tungsten is an important material for high-temperature applications due to its high chemical and thermal stability. Its carbide, that is, tungsten carbide, is used in tool manufacturing because of its outstanding hardness and as a catalyst scaffold due to its morphology and large surface area. However, microstructuring, especially high-resolution 3D microstructuring of both materials, is a complex and challenging process which suffers from slow speeds and requires expensive specialized equipment. Traditional subtractive machining methods, for example, milling, are often not feasible because of the hardness and brittleness of the materials. Commonly, tungsten and tungsten carbide are manufactured by powder metallurgy. However, these methods are very limited in the complexity and resolution of the produced components. Herein, tungsten ion-containing organic–inorganic photoresins, which are patterned by two-photon lithography (TPL) at micrometer resolution, are introduced. The printed structures are converted to tungsten or tungsten carbide by thermal debinding and reduction of the precursor or carbothermal reduction reaction, respectively. Using TPL, complex 3D tungsten and tungsten carbide structures are prepared with a resolution down to 2 and 7 μm, respectively. This new pathway of structuring tungsten and its carbide facilitates a broad range of applications from micromachining to metamaterials and catalysis.     

A novel predictive homotopic path tracking algorithm to solve non-linear algebraic equations

In chemical engineering and other areas of mathematics and engineering sciences, systems of non-linear algebraic equations often must be solved, and a problem is that these types of systems often cannot be solved analytically. Homotopic continuation methods are globally convergent and can find several solutions to the analyzed algebraic system and consist of a predictive step followed by a corrective step, which can cause reversion of homotopic path tracking. Therefore, in this work, the use of a homotopy continuation method (HCM) based on a purely predictive methodology is proposed; that is, it only makes use of predictor vectors (without correction vectors) to find the solution to several systems of non-linear algebraic equations with a multiplicity of solutions, employing lower central proccessing unit (CPU) time in comparison with standard homotopic tracking algorithms.