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Expression of ceramide anabolizing and catabolizing enzymes in MIN and CIN colorectal cells upon DL-PDMP treatment.

UGCG and SGMS1 knockdown efficiencies with different shRNA and siRNA.

DL-PDMP inhibits the proliferation of myeloid leukemia cell lines.

DL-PDMP does not affect proliferation of MEFs and colorectal cancer cells.

DL-PDMP treatment does not cause weight loss in mice.

Aneuploidy levels in Bub1b H/H and Cdc20 AAA/AAA MEFs.

Aneuploidy, a hallmark of cancer cells, poses an appealing opportunity for cancer treatment and prevention strategies. Using a cell-based screen to identify small molecules that could selectively kill aneuploid cells, we identified the compound N-[2-hydroxy-1-(4-morpholinylmethyl)-2-phenylethyl]-decanamide monohydrochloride (DL-PDMP), an antagonist of UDP-glucose ceramide glucosyltransferase. DL-PDMP selectively inhibited proliferation of aneuploid primary mouse embryonic fibroblasts and aneuploid colorectal cancer cells. Its selective cytotoxic effects were based on further accentuating the elevated levels of ceramide, which characterize aneuploid cells, leading to increased apoptosis. We observed that DL-PDMP could also enhance the cytotoxic effects of paclitaxel, a standard-of-care chemotherapeutic agent that causes aneuploidy, in human colon cancer and mouse lymphoma cells. Our results offer pharmacologic evidence that the aneuploid state in cancer cells can be targeted selectively for therapeutic purposes, or for reducing the toxicity of taxane-based drug regimens. Cancer Res; 77(19); 5272–86. ©2017 AACR.

Sphingolipid composition of aneuploid and euploid cell lines upon DL-PDMP treatment.

Table S1: Summary of compound effect in aneuploid Ts13 MEF compared to euploid wild-type MEF. Table S2: shRNA and siRNA sequences used in this study. Table S3: Primers used for quantitative Real-Time PCR.

Cobalt germanides have been widely studied as semiconductor contact materials, but recent theoretical studies suggest that they may also be excellent catalysts for methane steam reforming with stabilities and activities comparable to more expensive noble metal catalysts. We have sputter deposited CoGe alloy films and characterized their structure and morphology after post-deposition annealing in high vacuum up to 1000 °C. We used X-ray photoelectron spectroscopy to study the initial oxidation of amorphous and crystalline CoGe alloy surfaces under low pressures of O2 and H2O. The oxidation rate in O2 was found to be faster for an amorphous CoGe surface compared to a crystalline surface. We also found that there was little difference in the oxidation rate in H2O for either amorphous or crystalline surfaces. During O2 oxidation, the crystalline surface preferentially forms GeO and the amorphous surface preferentially forms GeO2. We have also observed preferential oxidation of Ge in the CoGe thin films. During temperature programmed desorption studies, we found that GeO desorption begins near 350 °C and that GeO2 decomposes to GeO and desorbs near 700 °C. More studies of CoGe catalysts are warranted, however GeO desorption may be a concern under reaction conditions when the film is subjected to an oxidizing environment.

Single cell biology has the potential to elucidate many critical biological processes and diseases, from development and regeneration to cancer. Single cell analyses are uncovering the molecular diversity of cells, revealing a clearer picture of the variation among and between different cell types. New techniques are beginning to unravel how differences in cell state-transcriptional, epigenetic, and other characteristics-can lead to different cell fates among genetically identical cells, which underlies complex processes such as embryonic development, drug resistance, response to injury, and cellular reprogramming. Single cell technologies also pose significant challenges relating to processing and analyzing vast amounts of data collected. To realize the potential of single cell technologies, new computational approaches are needed. On March 17-19, 2021, experts in single cell biology met virtually for the Keystone eSymposium "Single Cell Biology" to discuss advances both in single cell applications and technologies.

SUMMARYThe blood-brain barrier (BBB) is critical for protecting the brain and maintaining neuronal homeostasis. Although the BBB is a unique feature of the central nervous system (CNS) vasculature, not all brain regions have the same degree of impermeability. Differences in BBB permeability are important for controlling the local extracellular environment of specific brain regions to regulate the function and plasticity of particular neural circuits. However, how BBB heterogeneity occurs is poorly understood. Here, we demonstrate how regional specialization of the BBB is achieved. With unbiased cell profiling in small, defined brain regions, we compare the median eminence, which has a naturally leaky BBB, with the cortex, which has an impermeable BBB. We identify hundreds of molecular differences in endothelial cells (ECs) and demonstrate the existence of differences in perivascular astrocytes and pericytes in these regions, finding 3 previously unknown subtypes of astrocytes and several key differences in pericytes. By serial electron microscopy reconstruction and a novel, aqueous-based tissue clearing imaging method, we further reveal previously unknown anatomical specializations of these perivascular cells and their unique physical interactions with neighboring ECs. Finally, we identify ligand-receptor pairs between ECs and perivascular cells that may regulate regional BBB integrity in ECs. Using a bioinformatic approach we identified 26 and 26 ligand-receptor pairs underlying EC-pericyte and EC-astrocyte interactions, respectively. Our results demonstrate that differences in ECs, together with region-specific physical and molecular interactions with local perivascular cells, contribute to BBB functional heterogeneity. These regional cell inventories serve as a platform for further investigation of the dynamic and heterogeneous nature of the BBB in other brain regions. Identification of local BBB specializations provides insight into the function of different brain regions and will permit the development of region-specific drug delivery in the CNS.

The blood–brain barrier (BBB) restricts free access of molecules between the blood and the brain and is essential for regulating the neural microenvironment. Here, we describe how the BBB was initially characterized and how the current field evaluates barrier properties. We next detail the cellular nature of the BBB and discuss both the conservation and variation of BBB function across taxa. Finally, we examine our current understanding of mouse and zebrafish model systems, as we expect that comparison of the BBB across organisms will provide insight into the human BBB under normal physiological conditions and in neurological diseases.

A bounding risk assessment is presented that evaluates possible human health risk from a hypothetical scenario involving a 10,000-gallon release of flowback water from horizontal fracturing of Marcellus Shale. The water is assumed to be spilled on the ground, infiltrates into groundwater that is a source of drinking water, and an adult and child located downgradient drink the groundwater. Key uncertainties in estimating risk are given explicit quantitative treatment using Monte Carlo analysis. Chemicals that contribute significantly to estimated health risks are identified, as are key uncertainties and variables to which risk estimates are sensitive. The results show that hypothetical exposure via drinking water impacted by chemicals in Marcellus Shale flowback water, assumed to be spilled onto the ground surface, results in predicted bounds between 10-10 and 10-6 (for both adult and child receptors) for excess lifetime cancer risk. Cumulative hazard indices (HICUMULATIVE ) resulting from these hypothetical exposures have predicted bounds (5th to 95th percentile) between 0.02 and 35 for assumed adult receptors and 0.1 and 146 for assumed child receptors. Predicted health risks are dominated by noncancer endpoints related to ingestion of barium and lithium in impacted groundwater. Hazard indices above unity are largely related to exposure to lithium. Salinity taste thresholds are likely to be exceeded before drinking water exposures result in adverse health effects. The findings provide focus for policy discussions concerning flowback water risk management. They also indicate ways to improve the ability to estimate health risks from drinking water impacted by a flowback water spill (i.e., reducing uncertainty).

Only thin-film technology can satisfy the requirements of high spatial selectivity at high-channel-count electrode array designs by simultaneously good conformability to the targeted tissue through mechanical flexibility enriching future applications of functional neural stimulation. However, caused by the high impact of the microstructure on the mechanical and electrochemical film properties, varying fabrication processes of the same thin-film makes the difference between acute and chronic long-term stable electrodes. The influence of standard clinical electrical pulsing on flexible polyimide-based thin-film platinum electrodes for neuroprostheses, either sputter deposited or evaporated, and different diameters was assessed and compared. The electrochemical and morphological analysis showed a higher corrosion susceptibility and electrochemical degradation for the sputter deposited platinum electrodes with even total failures of smaller diameters. In contrast, the evaporated thin-films provided itself as more stable and reliable metallization with also smaller electrodes keeping their film integrity intact over the experimental period, -appearing to be the preferable material for improving thin-film electrodes' longevity.

The high complexity of the biological response to implanted materials builds a serious barrier against implanted recording and stimulation electrode arrays to succeed in clinically relevant chronic studies. Some of the cell and molecular interactions and their contribution to inflammation and device failure are still unclear. The interrelated mechanisms leading to tissue damage and electrode array failure during simultaneous faradaic, electrochemical reactions and biological response under electrical stimulation are not understood sufficiently. One variable, with which inflammatory and electrode surface processes can be analyzed and assessed, is the pH change in the immediate environment of the material-tissue interface. Here, the greatest challenges are in the biocompatibility and in-vivo long-term stability of selected sensor materials, the measurement of small transient pH oscillations and positioning of the sensor at a defined and nearest possible distance in the micrometer range, to the site of activity without the pH sensing being affected by the material- issue interactions itself. This work represents the in-situ measurement of local and transient pH changes at apulsed electrode with an embedded in-vivo compatible pH sensor and therein differentiating from current approaches of pH sensing during electrical stimulation.

The aim of the current study was to evaluate the effectiveness of intraoperative neuromonitoring (INM) as an adjunct in performing tarsal tunnel decompression surgery. We reviewed 38 patients who met inclusion criteria. INM was used to measure the voltage of the abductor hallucis and digiti quinti muscles both before and after decompression. Observed changes intraoperatively were acute and within minutes of the decompression performed by the surgeon. Patient outcomes were ascertained from clinical findings and classified as excellent, fair, or poor. Patient outcomes and the voltage change were measured and assessed for association, and statistically significant differences were found between outcome groups. Of the 38 patients, 29 (76%) had excellent outcomes, with a mean change in microvolts of 2088.28 ± 1172.44 (684%) (p = .0004) and 2173.24 ± 1228.39 (742%) (p = .0014) for abductor hallucis and abductor digiti quinti, respectively. The study supports INM as a useful adjunct in performing tarsal tunnel decompression.

Aneuploidy, a hallmark of cancer cells, poses an appealing opportunity for cancer treatment and prevention strategies. Using a cell-based screen to identify small molecules that could selectively kill aneuploid cells, we identified the compound N-[2hydroxy-1-(4-morpholinylmethyl)-2-phenylethyl]-decanamide monohydrochloride (DL-PDMP), an antagonist of UDP-glucose ceramide glucosyltransferase. DL-PDMP selectively inhibited proliferation of aneuploid primary mouse embryonic fibroblasts and aneuploid colorectal cancer cells. Its selective cytotoxic effects were based on further accentuating the elevated levels of ceramide, which characterize aneuploid cells, leading to increased apoptosis. We observed that DL-PDMP could also enhance the cytotoxic effects of paclitaxel, a standard-of-care chemotherapeutic agent that causes aneuploidy, in human colon cancer and mouse lymphoma cells. Our results offer pharmacologic evidence that the aneuploid state in cancer cells can be targeted selectively for therapeutic purposes, or for reducing the toxicity of taxane-based drug regimens., National Cancer Institute (U.S.) (Grant P30-CA14051)

Miniaturization of electrodes is a prerequisite of selective and targeted interaction with single neurons, enabling more applications in the continuously growing field of neuroprostheses. Miniaturization in all three dimensions of the electrical contact sites should maintain or increase longevity and electrical functionality. The thin-film metallization of the electrode site, which is only a couple of hundreds of nanometers thick, has to withstand high chemical load through the corrosive environment in the body and the electrochemical processes during electrical stimulation in vivo. Platinum (Pt), which is known to be chemically inert and mechanical stable as bulk material shows a lack of chemical and mechanical integrity applied in thin-film microelectrodes. In our study we investigated failure mechanisms of thin-film Pt electrodes under conditions of electrode aging and electrical stimulation in different physiological media. To understand and eventually overcome stability loss, we investigated the intrinsic structural stress and deformations that arose from mechanical loading through chemical impact and electrical stimulation using optical microscopy and white-light interferometry. Electrochemical measurements indicated oxidation and surface roughening as two of the degradation processes in thin-film electrodes. From the results presumptions about the underlying microstructural changes were made.

A bounding risk assessment is presented that evaluates possible human health risk from a hypothetical scenario involving a 10,000-gallon release of flowback water from horizontal fracturing of Marcellus Shale. The water is assumed to be spilled on the ground, infiltrates into groundwater that is a source of drinking water, and an adult and child located downgradient drink the groundwater. Key uncertainties in estimating risk are given explicit quantitative treatment using Monte Carlo analysis. Chemicals that contribute significantly to estimated health risks are identified, as are key uncertainties and variables to which risk estimates are sensitive. The results show that hypothetical exposure via drinking water impacted by chemicals in Marcellus Shale flowback water, assumed to be spilled onto the ground surface, results in predicted bounds between 10

A method for determining a safety range for non-cancer risks is proposed, similar in concept to the range used for cancer in the management of waste sites. This safety range brings transparency to the chemical specific Reference Dose or Concentration by replacing their “order of magnitude” definitions with a scientifically-based range. EPA’s multiple RfCs for trichloroethylene (TCE) were evaluated as a case study. For TCE, a multi-endpoint safety range was judged to be 3 μg/m3 to 30 μg/m,3 based on a review of kidney effects found in NTP (1988), thymus effects found in Keil et al. (2009) and cardiac effects found in the Johnson et al. (2003) study. This multi-endpoint safety range is derived from studies for which the appropriate averaging time corresponds to different exposure durations, and, therefore, can be applied to both long- and short-term exposures with appropriate consideration of exposure averaging times. For shorter-term exposures, averaging time should be based on the time of cardiac development in humans during fetal growth, an average of approximately 20–25 days.

Aneuploidy, an imbalanced chromosome number, is associated with both cancer and developmental disorders such as Down syndrome (DS). To determine how aneuploidy affects cellular and organismal physiology, we have developed a system to evaluate aneuploid cell fitness in vivo. By transplanting hematopoietic stem cells (HSCs) into recipient mice after ablation of recipient hematopoiesis by lethal irradiation, we can directly compare the fitness of HSCs derived from a range of aneuploid mouse models with that of euploid HSCs. This experimental system can also be adapted to assess the interplay between aneuploidy and tumorigenesis. We hope that further characterization of aneuploid cells in vivo will provide insight both into the origins of hematopoietic phenotypes observed in DS individuals as well as the role of different types of aneuploid cells in the genesis of cancers of the blood., National Institutes of Health (U.S.) (Grant GM056800)

Sintering of metal nanowire (NW) networks on transparent polymers is an emerging approach for fabricating transparent conductive electrodes used in multiple devices. Pulsed light sintering is a scalable sintering process in which large-area, broad-spectrum xenon lamp light causes rapid NW fusion to increase network conductivity, while embedding the NWs in the polymer to increase mechanical robustness. This paper develops a multiphysical approach for predicting evolution of conductivity, NW fusion and nanoscale temperature gradients on the substrate during pulsed light sintering of silver NWs on polycarbonate. Model predictions are successfully validated against experimentally measured temperature and electrical resistance evolution. New insight is obtained into the diameter-dependent kinetics of NW fusion and nanoscale temperature gradients on the substrate, which are difficult to obtain experimentally. These observations also lead to the understanding that NW embedding in intense pulsed light sintering (IPL) can occur below the glass transition temperature of the polymer, and to a new differential thermal expansion-based mechanism of NW embedding during IPL. These insights, and the developed model, create a framework for physics-guided choice of NWs, substrate and process parameters to control conductivity and prevent substrate damage during the process.

Recent studies have indicated that nickel gallium alloys can be effective catalysts for the hydrogenation of CO2 to methanol. To simplify the characterization of NiGa catalysts, the authors are developing model systems using sputter deposited NiGa thin films. The NiGa thin films used in this study were deposited using an equimolar alloy target and annealed in ultrahigh vacuum. Atomic force microscopy (AFM), x-ray diffraction (XRD), and x-ray photoelectron spectroscopy (XPS) were used to characterize the NiGa films before and after annealing. AFM results showed that film roughness and grain size significantly increased as the film was annealed above 700 °C. XRD patterns indicated that NiGa thin films were nanocrystalline as deposited and then transitioned to the Ni13Ga9 phase after annealing above 500 °C. XPS results indicated that gallium and oxygen segregated to the surface after annealing up to 600 °C, and formed a surface Ga2O3 layer. For anneals above 600 °C, the Ga2O3 XPS signal was reduced in intens...

Aneuploidy, an imbalanced karyotype, is a widely observed feature of cancer cells that has long been hypothesized to promote tumorigenesis. Here we evaluate the fitness of cells with constitutional trisomy or chromosomal instability (CIN) in vivo using hematopoietic reconstitution experiments. We did not observe cancer but instead found that aneuploid hematopoietic stem cells (HSCs) exhibit decreased fitness. This reduced fitness is due at least in part to the decreased proliferative potential of aneuploid hematopoietic cells. Analyses of mice with CIN caused by a hypomorphic mutation in the gene Bub1b further support the finding that aneuploidy impairs cell proliferation in vivo. Whereas nonregenerating adult tissues are highly aneuploid in these mice, HSCs and other regenerative adult tissues are largely euploid. These findings indicate that, in vivo, mechanisms exist to select against aneuploid cells., National Institutes of Health (U.S.) (CA206157), Kathy and Curt Marble Cancer Research Fund, David H. Koch Institute for Integrative Cancer Research at MIT (Support Grant P30-CA14051), National Institute of General Medical Sciences (U.S.) (Training Grant T32GM007753)