Sensitization of renal carcinoma cells to TRAIL-induced apoptosis by rocaglamide and analogs

Rocaglamide has been reported to sensitize several cell types to TRAIL-induced apoptosis. In recent years, advances in synthetic techniques have led to generation of novel rocaglamide analogs. However, these have not been extensively analyzed as TRAIL sensitizers, particularly in TRAIL-resistant renal cell carcinoma cells. Evaluation of rocaglamide and analogs identified 29 compounds that are able to sensitize TRAIL-resistant ACHN cells to TRAIL-induced, caspase-dependent apoptosis with sub-µM potency which correlated with their potency as protein synthesis inhibitors and with loss of cFLIP protein in the same cells. Rocaglamide alone induced cell cycle arrest, but not apoptosis. Rocaglates averaged 4–5-fold higher potency as TRAIL sensitizers than as protein synthesis inhibitors suggesting a potential window for maximizing TRAIL sensitization while minimizing effects of general protein synthesis inhibition. A wide range of other rocaglate effects (e.g. on JNK or RAF-MEK-ERK signaling, death receptor levels, ROS, ER stress, eIF4E phosphorylation) were assessed, but did not contribute to TRAIL sensitization. Other than a rapid loss of MCL-1, rocaglates had minimal effects on mitochondrial apoptotic pathway proteins. The identification of structurally diverse/mechanistically similar TRAIL sensitizing rocaglates provides insights into both rocaglate structure and function and potential further development for use in RCC-directed combination therapy.This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. This research was supported [in part] by the Intramural Research Program of NIH, Frederick. National Lab, Center for Cancer Research. Research performed at Boston University was supported in part by NIH R35 GM118173. Work at the BU-CMD is supported by R24 GM111625. (HHSN261200800001E - National Cancer Institute, National Institutes of Health; Intramural Research Program of NIH, Frederick. National Lab, Center for Cancer Research; R35 GM118173 - NIH; R24 GM111625)Published versio

Satellite observed water quality changes in the Laurentian Great Lakes due to invasive species, anthropogenic forcing, and climate change

Long time series of ocean and land color satellite data can be used to measure Laurentian Great Lakes water quality parameters including chlorophyll, suspended minerals, harmful algal blooms (HABs), photic zone and primary productivity on weekly, monthly and annual observational intervals. The observed changes in these water quality parameters over time are a direct result of the introduction of invasive species such as the Dreissena mussels as well as anthropogenic forcing and climate change. Time series of the above mentioned water quality parameters have been generated based on a range of satellite sensors, starting with Landsat in the 1970s and continuing to the present with MODIS and VIIRS. These time series have documented the effect the mussels have had on increased water clarity by decreasing the chlorophyll concentrations. Primary productivity has declined in the lakes due to the decrease in algae. The increased water clarity due to the mussels has also led to an increase in submerged aquatic vegetation. Comparing water quality metrics in Lake Superior to the lower lakes is insightful because Lake Superior is the largest and most northern of the five Great Lakes and to date has not been affected by the invasive mussels and can thus be considered a control. In contrast, Lake Erie, the most southern and shallow of the Laurentian Great Lakes, is heavily influenced by agricultural practices (i.e., nutrient runoff) and climate change, which directly influence the annual extent of HABs in the Western Basin of that lake

Galaxy Cluster Scaling Relations between Bolocam Sunyaev-Zel'dovich Effect and Chandra X-ray Measurements

We present scaling relations between the integrated Sunyaev-Zel'dovich Effect (SZE) signal, $Y_{\rm SZ}$, its X-ray analogue, $Y_{\rm X}\equiv M_{\rm gas}T_{\rm X}$, and total mass, $M_{\rm tot}$, for the 45 galaxy clusters in the Bolocam X-ray-SZ (BOXSZ) sample. All parameters are integrated within $r_{2500}$. $Y_{2500}$ values are measured using SZE data collected with Bolocam, operating at 140 GHz at the Caltech Submillimeter Observatory (CSO). The temperature, $T_{\rm X}$, and mass, $M_{\rm gas,2500}$, of the intracluster medium are determined using X-ray data collected with Chandra, and $M_{\rm tot}$ is derived from $M_{\rm gas}$ assuming a constant gas mass fraction. Our analysis accounts for several potential sources of bias, including: selection effects, contamination from radio point sources, and the loss of SZE signal due to noise filtering and beam-smoothing effects. We measure the $Y_{2500}$--$Y_{\rm X}$ scaling to have a power-law index of $0.84\pm0.07$, and a fractional intrinsic scatter in $Y_{2500}$ of $(21\pm7)\%$ at fixed $Y_{\rm X}$, both of which are consistent with previous analyses. We also measure the scaling between $Y_{2500}$ and $M_{2500}$, finding a power-law index of $1.06\pm0.12$ and a fractional intrinsic scatter in $Y_{2500}$ at fixed mass of $(25\pm9)\%$. While recent SZE scaling relations using X-ray mass proxies have found power-law indices consistent with the self-similar prediction of 5/3, our measurement stands apart by differing from the self-similar prediction by approximately 5$\sigma$. Given the good agreement between the measured $Y_{2500}$--$Y_{\rm X}$ scalings, much of this discrepancy appears to be caused by differences in the calibration of the X-ray mass proxies adopted for each particular analysis.Comment: 31 pages, 15 figures, accepted by ApJ 04/11/2015. This version is appreciably different from the original submission: it includes an entirely new appendix, extended discussion, and much of the material has been reorganize

Satellite monitoring of harmful algal blooms in the Western Basin of Lake Erie: A 20-year time-series

Blooms of harmful cyanobacteria (cyanoHABs) have occurred on an annual basis in western Lake Erie for more than a decade. Previously, we developed and validated an algorithm to map the extent of the submerged and surface scum components of cyanoHABs using MODIS ocean-color satellite data. The algorithm maps submerged cyanoHABs by identifying high chlorophyll concentrations (\u3e18 mg/m3) combined with water temperature \u3e20 °C, while cyanoHABs surface scums are mapped using near-infrared reflectance values. Here, we adapted this algorithm for the SeaWiFS sensor to map the annual areal extents of cyanoHABs in the Western Basin of Lake Erie for the 20-year period from 1998 to 2017. The resulting classified maps were validated by comparison with historical in situ measurements, exhibiting good agreement (81% accuracy). Trends in the annual mean and maximum total submerged and surface scum extents demonstrated significant positive increases from 1998 to 2017. There was also an apparent 76% increase in year-to-year variability of mean annual extent between the 1998–2010 and 2011–2017 periods. The 1998–2017 time-series was also compared with several different river discharge nutrient loading metrics to assess the ability to predict annual cyanoHAB extents. The prediction models displayed significant relationships between spring discharge and cyanoHAB area; however, substantial variance remained unexplained due in part to the presence of very large blooms occurring in 2013 and 2015. This new multi-sensor time-series and associated statistics extend the current understanding of the extent, location, duration, and temporal patterns of cyanoHABs in western Lake Erie

A model for transition of 5 '-nuclease domain of DNA polymerase I from inert to active modes

Bacteria contain DNA polymerase I (PolI), a single polypeptide chain consisting of similar to 930 residues, possessing DNA-dependent DNA polymerase, 3'-5' proofreading and 5'-3' exonuclease (also known as flap endonuclease) activities. PolI is particularly important in the processing of Okazaki fragments generated during lagging strand replication and must ultimately produce a double-stranded substrate with a nick suitable for DNA ligase to seal. PolI's activities must be highly coordinated both temporally and spatially otherwise uncontrolled 5'-nuclease activity could attack a nick and produce extended gaps leading to potentially lethal double-strand breaks. To investigate the mechanism of how PolI efficiently produces these nicks, we present theoretical studies on the dynamics of two possible scenarios or models. In one the flap DNA substrate can transit from the polymerase active site to the 5'-nuclease active site, with the relative position of the two active sites being kept fixed; while the other is that the 5'-nuclease domain can transit from the inactive mode, with the 5'-nuclease active site distant from the cleavage site on the DNA substrate, to the active mode, where the active site and substrate cleavage site are juxtaposed. The theoretical results based on the former scenario are inconsistent with the available experimental data that indicated that the majority of 5'-nucleolytic processing events are carried out by the same PolI molecule that has just extended the upstream primer terminus. By contrast, the theoretical results on the latter model, which is constructed based on available structural studies, are consistent with the experimental data. We thus conclude that the latter model rather than the former one is reasonable to describe the cooperation of the PolI's polymerase and 5'-3' exonuclease activities. Moreover, predicted results for the latter model are presented

Spatial and temporal variability of inherent and apparent optical properties in western Lake Erie: Implications for water quality remote sensing

Lake Erie has experienced dramatic changes in water quality over the past several decades requiring extensive monitoring to assess effectiveness of adaptive management strategies. Remote sensing offers a unique potential to provide synoptic monitoring at daily time scales complementing in-situ sampling activities occurring in Lake Erie. Bio-optical remote sensing algorithms require knowledge about the inherent optical properties (IOPs) of the water for parameterization to produce robust water quality products. This study reports new IOP and apparent optical property (AOP) datasets for western Lake Erie that encapsulate the May–October period for 2015 and 2016 at weekly sampling intervals. Previously reported IOP and AOP observations have been temporally limited and have not assessed statistical differences between IOPs over spatial and temporal gradients. The objective of this study is to assess trends in IOPs over variable spatial and temporal scales. Large spatio-temporal variability in IOPs was observed between 2015 and 2016 likely due to the difference in the extent and duration of mid-summer cyanobacteria blooms. Differences in the seasonal trends of the specific phytoplankton absorption coefficient between 2015 and 2016 suggest differing algal assemblages between the years. Other IOP variables, including chromophoric, dissolved organic matter (CDOM) and beam attenuation spectral slopes, suggest variability is influenced by river discharge and sediment re-suspension. The datasets presented in this study show how these IOPs and AOPs change over a season and between years, and are useful in advancing the applicability and robustness of remote sensing methods to retrieve water quality information in western Lake Erie

Genetic variability in the humoral immune response to bovine herpesvirus-1 infection in dairy cattle and genetic correlations with performance traits

peer-reviewedBovine herpesvirus-1 (BoHV-1) is a viral pathogen of global significance that is known to instigate several diseases in cattle, the most notable of which include infectious bovine rhinotracheitis and bovine respiratory disease. The genetic variability in the humoral immune response to BoHV-1 has, to our knowledge, not ever been quantified. Therefore, the objectives of the present study were to estimate the genetic parameters for the humoral immune response to BoHV-1 in Irish female dairy cattle, as well as to investigate the genetic relationship between the humoral immune response to BoHV-1 with milk production performance, fertility performance, and animal mortality. Information on antibody response to BoHV-1 was available to the present study from 2 BoHV-1 sero-prevalence research studies conducted between the years 2010 to 2015, inclusive; after edits, BoHV-1 antibody test results were available on a total of 7,501 female cattle from 58 dairy herds. National records of milk production (i.e., 305-d milk yield, fat yield, protein yield, and somatic cell score; n = 1,211,905 milk-recorded cows), fertility performance (i.e., calving performance, pregnancy diagnosis, and insemination data; n = 2,365,657 cows) together with animal mortality data (i.e., birth, farm movement, death, slaughter, and export events; n = 12,853,257 animals) were also available. Animal linear mixed models were used to quantify variance components for BoHV-1 as well as to estimate genetic correlations among traits. The estimated genetic parameters for the humoral immune response to BoHV-1 in the present study (i.e., heritability range: 0.09 to 0.16) were similar to estimates previously reported for clinical signs of bovine respiratory disease in dairy and beef cattle (i.e., heritability range: 0.05 to 0.11). Results from the present study suggest that breeding for resistance to BoHV-1 infection could reduce the incidence of respiratory disease in cattle while having little or no effect on genetic selection for milk yield or milk constituents (i.e., genetic correlations ranged from −0.13 to 0.17). Moreover, even though standard errors were large, results also suggest that breeding for resistance to BoHV-1 infection may indirectly improve fertility performance while also reducing the incidence of mortality in older animals (i.e., animals >182 d of age). Results can be used to inform breeding programs of potential genetic gains achievable for resistance to BoHV-1 infection in cattle.Funding from the Irish Department of Agriculture, Food and the Marine STIMULUS research grant HealthyGenes (Dublin

A Fluctuation Analysis of the Bolocam 1.1mm Lockman Hole Survey

We perform a fluctuation analysis of the 1.1mm Bolocam Lockman Hole Survey, which covers 324 square arcmin to a very uniform point source-filtered RMS noise level of 1.4 mJy/beam. The fluctuation analysis has the significant advantage of utilizing all of the available data. We constrain the number counts in the 1-10 mJy range, and derive significantly tighter constraints than in previous work: the power-law index is 2.7 (+0.18, -0.15), while the amplitude is equal to 1595 (+85,-238) sources per mJy per square degree, or N(>1 mJy) = 940 (+50,-140) sources/square degree (95% confidence). Our results agree extremely well with those derived from the extracted source number counts by Laurent et al (2005). Our derived normalization is about 2.5 times smaller than determined by MAMBO at 1.2mm by Greve et al (2004). However, the uncertainty in the normalization for both data sets is dominated by the systematic (i.e., absolute flux calibration) rather than statistical errors; within these uncertainties, our results are in agreement. We estimate that about 7% of the 1.1mm background has been resolved at 1 mJy.Comment: To appear in the Astrophysical Journal; 22 pages, 9 figure

Frontier fields clusters: deep Chandra observations of the complex merger MACS~J1149.6+2223

The Hubble Space Telescope Frontier Fields cluster MACS J1149.6+2223 is one of the most complex merging clusters, believed to consist of four dark matter halos. We present results from deep (365 ks) Chandra observations of the cluster, which reveal the most distant cold front (z = 0.544) discovered to date. In the cluster outskirts, we also detect hints of a surface brightness edge that could be the bow shock preceding the cold front. The substructure analysis of the cluster identified several components with large relative radial velocities, thus indicating that at least some collisions occur almost along the line of sight. The inclination of the mergers with respect to the plane of the sky poses significant observational challenges at X-ray wavelengths. MACS J1149.6+2223 possibly hosts a steep-spectrum radio halo. If the steepness of the radio halo is confirmed, then the radio spectrum, combined with the relatively regular ICM morphology, could indicate that MACS J1149.6+2223 is an old merging cluster

Evaluating the cytotoxicity of innate immune effector cells using the GrB ELISPOT assay

BACKGROUND: This study assessed the Granzyme B (GrB) ELISPOT as a viable alternative to the (51)Cr-release assay for measuring cytotoxic activity of innate immune effector cells. We strategically selected the GrB ELISPOT assay because GrB is a hallmark effector molecule of cell-mediated destruction of target cells. METHODS: We optimized the GrB ELISPOT assay using the human-derived TALL-104 cytotoxic cell line as effectors against K562 target cells. Titration studies were performed to assess whether the ELISPOT assay could accurately enumerate the number of GrB-secreting effector cells. TALL-104 were treated with various secretion inhibitors and utilized in the GrB ELISPOT to determine if GrB measured in the ELISPOT was due to degranulation of effector cells. Additionally, CD107a expression on effector cells after effector-target interaction was utilized to further confirm the mechanism of GrB release by TALL-104 and lymphokine-activated killer (LAK) cells. Direct comparisons between the GrB ELISPOT, the IFN-γ ELISPOT and the standard (51)Cr-release assays were made using human LAK cells. RESULTS: Titration studies demonstrated a strong correlation between the number of TALL-104 and LAK effector cells and the number of GrB spots per well. GrB secretion was detectable within 10 min of effector-target contact with optimal secretion observed at 3–4 h; in contrast, optimal IFN-γ secretion was not observed until 24 h. The protein secretion inhibitor, brefeldin A, did not inhibit the release of GrB but did abrogate IFN-γ production by TALL-104 cells. GrB secretion was abrogated by BAPTA-AM (1,2-bis-(2-aminophenoxy)ethane-N,N,N', N'-tetraacetic acid tetra(acetoxymethyl) ester), which sequesters intracellular Ca(2+), thereby preventing degranulation. The number of effector cells expressing the degranulation associated glycoprotein CD107a increased after interaction with target cells and correlated with the stimulated release of GrB measured in the ELISPOT assay. CONCLUSIONS: Because of its high sensitivity and ability to estimate cytotoxic effector cell frequency, the GrB ELISPOT assay is a viable alternative to the (51)Cr-release assay to measure MHC non-restricted cytotoxic activity of innate immune cells. Compared to the IFN-γ ELISPOT assay, the GrB ELISPOT may be a more direct measure of cytotoxic cell activity. Because GrB is one of the primary effector molecules in natural killer (NK) cell-mediated killing, detection and enumeration of GrB secreting effector cells can provide valuable insight with regards to innate immunological responses