Probing Dynein and Kinesin Stepping with Mechanical Manipulation in a Living Cell

Molecular motors: By combining optical tweezers and high-speed particle tracking, individual steps of microtubule motor proteins transporting organelles can be detected under known force loads in living mammalian cells (see figure). We report a label-free assay for simultaneous optical manipulation and tracking of endogenous lipid droplets as actively transported cargoes in a living mammalian cell with sub-millisecond time resolution. Using an EM-CCD camera as a highly sensitive quadrant detector, we can detect steps of dynein- and kinesin-driven cargoes under known force loads. We can distinguish single and multiple motor-driven cargoes and show that the stall forces for inward and outward transported cargoes are similar. By combining the stall force observable with the ability to detect individual steps, we can characterize kinesin- and dynein-driven active transport in different force regimes.Chemistry and Chemical Biolog

FAK promotes stromal PD-L2 expression associated with poor survival in pancreatic cancer

BACKGROUND: Pancreatic Cancer is one of the most lethal cancers, with less than 8% of patients surviving 5 years following diagnosis. The last 40 years have seen only small incremental improvements in treatment options, highlighting the continued need to better define the cellular and molecular pathways contributing to therapy response and patient prognosis. METHODS: We combined CRISPR, shRNA and flow cytometry with mechanistic experiments using a Kras(G12D)p53(R172H) mouse model of pancreatic cancer and analysis of publicly available human PDAC transcriptomic datasets. RESULTS: Here, we identify that expression of the immune checkpoint, Programmed Death Ligand 2 (PD-L2), is associated with poor prognosis, tumour grade, clinical stage and molecular subtype in patients with Pancreatic Ductal Adenocarcinoma (PDAC). We further show that PD-L2 is predominantly expressed in the stroma and, using an orthotopic murine model of PDAC, identify cancer cell-intrinsic Focal Adhesion Kinase (FAK) signalling as a regulator of PD-L2 stromal expression. Mechanistically, we find that FAK regulates interleukin-6, which can act in concert with interleukin-4 secreted by CD4 T-cells to drive elevated expression of PD-L2 on tumour-associated macrophages, dendritic cells and endothelial cells. CONCLUSIONS: These findings identify further complex heterocellular signalling networks contributing to FAK-mediated immune suppression in pancreatic cancer

Alder Canopy Dieback and Damage in Western Oregon Riparian Ecosystems

We gathered baseline data to assess alder tree damage in western Oregon riparian ecosystems. We sought to determine if Phytophthora-type cankers found in Europe or the pathogen Phytophthora alni subsp. alni, which represent a major threat to alder forests in the Pacific Northwest, were present in the study area. Damage was evaluated in 88 transects; information was recorded on damage type (pathogen, insect or wound) and damage location. We evaluated 1445 red alder (Alnus rubra), 682 white alder (Alnus rhombifolia) and 181 thinleaf alder (Alnus incana spp. tenuifolia) trees. We tested the correlation between canopy dieback and canker symptoms because canopy dieback is an important symptom of Phytophthora disease of alder in Europe. We calculated the odds that alder canopy dieback was associated with Phytophthora-type cankers or other biotic cankers. P. alni subsp. alni (the causal agent of alder disease in Europe) was not identified in western Oregon; however, Phytophthora siskiyouensis was isolated from Phytophthora-type cankers which were present on 2% of red alder trees and 3% of white alder trees. The odds of canopy dieback were 5.4 and 4.8 times greater for red and white alder (respectively) with Phytophthora-type canker symptoms than in trees without such cankers. The percentage of trees with canopy dieback was 51%, 32%, and 10% for red, white, and thinleaf alder respectively. Other common damage included wounding, foliar pathogens and insects on red alder. This is the first report of Phytophthora canker of alder in United States forests and first report of P. siskiyouensis isolation from alder in forests anywhere.Keywords: alder tree damage, Phytophthora species, riparian ecosystems, symptom

Nuclear FAK and Runx1 cooperate to regulate IGFBP3, cell cycle progression and tumor growth

Abstract Nuclear focal adhesion kinase (FAK) is a potentially important regulator of gene expression in cancer, impacting both cellular function and the composition of the surrounding tumor microenvironment. Here, we report in a murine model of skin squamous cell carcinoma (SCC) that nuclear FAK regulates Runx1-dependent transcription of insulin-like growth factor binding protein 3 (IGFBP3), and that this regulates SCC cell-cycle progression and tumor growth in vivo. Furthermore, we identified a novel molecular complex between FAK and Runx1 in the nucleus of SCC cells and showed that FAK interacted with a number of Runx1-regulatory proteins, including Sin3a and other epigenetic modifiers known to alter Runx1 transcriptional function through posttranslational modification. These findings provide important new insights into the role of FAK as a scaffolding protein in molecular complexes that regulate gene transcription. Cancer Res; 77(19); 5301–12. ©2017 AACR.</jats:p

A Bayesian Calibration Framework for EDGES

We develop a Bayesian model that jointly constrains receiver calibration, foregrounds and cosmic 21cm signal for the EDGES global 21\,cm experiment. This model simultaneously describes calibration data taken in the lab along with sky-data taken with the EDGES low-band antenna. We apply our model to the same data (both sky and calibration) used to report evidence for the first star formation in 2018. We find that receiver calibration does not contribute a significant uncertainty to the inferred cosmic signal (<1%), though our joint model is able to more robustly estimate the cosmic signal for foreground models that are otherwise too inflexible to describe the sky data. We identify the presence of a significant systematic in the calibration data, which is largely avoided in our analysis, but must be examined more closely in future work. Our likelihood provides a foundation for future analyses in which other instrumental systematics, such as beam corrections and reflection parameters, may be added in a modular manner.Comment: 18 pages + 3 for appendices. 13 figures. Accepted to MNRA

Analytic approximations of scattering effects on beam chromaticity in 21-cm global experiments

Scattering from objects near an antenna produce correlated signals from strong compact radio sources in a manner similar to those used by the Sea Interferometer to measure the radio source positions using the fine frequency structure in the total power spectrum of a single antenna. These fringes or ripples due to correlated signal interference are present at a low level in the spectrum of any single antenna and are a major source of systematics in systems used to measure the global redshifted 21-cm signal from the early universe. In the Sea Interferometer a single antenna on a cliff above the sea is used to add the signal from the direct path to the signal from the path reflected from the sea thereby forming an interferometer. This was used for mapping radio sources with a single antenna by Bolton and Slee in the 1950s. In this paper we derive analytic expressions to determine the level of these ripples and compare these results in a few simple cases with electromagnetic modeling software to verify that the analytic calculations are sufficient to obtain the magnitude of the scattering effects on the measurements of the global 21-cm signal. These analytic calculations are needed to evaluate the magnitude of the effects in cases that are either too complex or take too much time to be modeled using software