Time-resolved X-ray diffraction diagnostic development for the National Ignition Facility.

We present the development of an experimental platform that can collect four frames of x-ray diffraction data along a single line of sight during laser-driven, dynamic-compression experiments at the National Ignition Facility. The platform is comprised of a diagnostic imager built around ultrafast sensors with a 2-ns integration time, a custom target assembly that serves also to shield the imager, and a 10-ns duration, quasi-monochromatic x-ray source produced by laser-generated plasma. We demonstrate the performance with diffraction data for Pb ramp compressed to 150 GPa and illuminated by a Ge x-ray source that produces ∼7 × 1011, 10.25-keV photons/ns at the 400 µm diameter sample.

Analysis and mitigation of an oscillating background on hybrid complementary metal-oxide semiconductor (hCMOS) imaging sensors at the National Ignition Facility.

Nanosecond-gated hybrid complementary metal-oxide semiconductor imaging sensors are a powerful tool for temporally gated and spatially resolved measurements in high energy density science, including inertial confinement fusion, and in laser diagnostics. However, a significant oscillating background excited by photocurrent has been observed in image sequences during testing and in experiments at the National Ignition Facility (NIF). Characterization measurements and simulation results are used to explain the oscillations as the convolution of the pixel-level sensor response with a sensor-wide RLC circuit ringing. Data correction techniques are discussed for NIF diagnostics, and for diagnostics where these techniques cannot be used, a proof-of-principle image correction algorithm is presented.

High-content microscopy reveals a morphological signature of bortezomib resistance.

Drug resistance is a challenge in anticancer therapy, particularly with targeted therapeutics and cytotoxic compounds. In many cases, cancers can be resistant to the drug prior to exposure, i.e., possess intrinsic drug resistance. However, we lack target-independent methods to anticipate resistance in cancer cell lines or characterize intrinsic drug resistance without a priori knowledge of its cause. We hypothesized that cell morphology could provide an unbiased readout of drug sensitivity prior to treatment. We therefore isolated clonal cell lines that were either sensitive or resistant to bortezomib, a well-characterized proteasome inhibitor and anticancer drug to which many cancer cells possess intrinsic resistance. We then measured high-dimensional single-cell morphology profiles using Cell Painting, a high-content microscopy assay. Our imaging- and computation-based profiling pipeline identified morphological features typically different between resistant and sensitive clones. These features were compiled to generate a morphological signature of bortezomib resistance, which correctly predicted the bortezomib treatment response in seven of ten cell lines not included in the training dataset. This signature of resistance was specific to bortezomib over other drugs targeting the ubiquitin-proteasome system. Our results provide evidence that intrinsic morphological features of drug resistance exist and establish a framework for their identification.

A 2-4 keV multilayer mirrored channel for the NIF Dante system.

During inertial confinement fusion experiments at the National Ignition Facility (NIF), a capsule filled with deuterium and tritium (DT) gas, surrounded by a DT ice layer and a high-density carbon ablator, is driven to the temperature and densities required to initiate fusion. In the indirect method, 2 MJ of NIF laser light heats the inside of a gold hohlraum to a radiation temperature of 300 eV; thermal x rays from the hohlraum interior couple to the capsule and create a central hotspot at tens of millions degrees Kelvin and a density of 100-200 g/cm3. During the laser interaction with the gold wall, m-band x rays are produced at ∼2.5 keV; these can penetrate into the capsule and preheat the ablator and DT fuel. Preheat can impact instability growth rates in the ablation front and at the fuel-ablator interface. Monitoring the hohlraum x-ray spectrum throughout the implosion is, therefore, critical; for this purpose, a Multilayer Mirror (MLM) with flat response in the 2-4 keV range has been installed in the NIF 37° Dante calorimeter. Precision engineering and x-ray calibration of components mean the channel will report 2-4 keV spectral power with an uncertainty of ±8.7%.

Design of the high-yield time-gated x-ray hot-spot imager for OMEGA.

Time-resolved x-ray self-emission imaging of hot spots in inertial confinement fusion experiments along several lines of sight provides critical information on the pressure and the transient morphology of the hot spot on the University of Rochester's OMEGA Laser System. At least three quasi-orthogonal lines of sight are required to infer the tomographic information of the hot spots of deuterium-tritium cryogenic layered implosions. OMEGA currently has two time-gated x-ray hot-spot imagers: the time-resolved Kirkpatrick-Baez x-ray microscope and the single-line-of-sight, time-resolved x-ray imager (SLOS-TRXI). The time-gated x-ray hot-spot imager (XRHSI) is being developed for use on OMEGA as the third line of sight for the high-yield operation of up to 4 × 1014 neutrons. XRHSI follows the SLOS-TRXI concept; however, it will have improved spatial and temporal resolutions of 5 µm and 20 ps, respectively. The simultaneous operation of the three instruments will provide 3-D reconstructions of the assembled hot-spot fuel at various times through peak thermonuclear output. The technical approach consists of a pinhole array imager and demagnifying time-dilation drift tube that are coupled to two side-by-side hybrid complementary metal-oxide semiconductor (hCMOS) image sensors. To minimize the background and to harden the diagnostics, an angled drift-tube assembly shifting the hCMOS sensors out of the direct line of sight and neutron shielding will be applied. The technical design space for the instrument will be discussed and the conceptual design will be presented.

Characterization of the hardened single line of sight camera at the National Ignition Facility.

The hardened single line of sight camera has been recently characterized in preparation for its deployment on the National Ignition Facility. The latest creation based on the pulse-dilation technology leads to many new features and improvements over the previous-generation cameras to provide better quality measurements of inertial confinement fusion experiments, including during high neutron yield implosions. Here, we present the characterization data that illustrate the main performance features of this instrument, such as extended dynamic range and adjustable internal magnification, leading to improved spatial resolution.

Compatibility and stability of an admixture of multiple anaesthetic drugs for opioid-free anaesthesia.

The ability to combine and use drugs in a single infusion device may be useful in resource-limited settings. This study examined the chemical stability of an opioid-sparing mixture of ketamine, lidocaine and magnesium sulphate when combined in a single syringe. High-performance liquid chromatography and atomic absorption spectrophotometry were performed on six syringes containing the three-drug mixture. Since most opioid-sparing techniques typically rely on a 24-hour infusion regime, we tested stability at the initial admixing and 24 hours later. Stability was defined as a measured drug concentration within 10% of expected, with the absence of precipitation or pH alterations. Pharmacokinetic simulations were conducted to further show that the achieved plasma drug concentrations were well within an effective analgesic range. All mixed drug concentration measurements were within the required 10% reference limit. No obvious precipitation or interaction occurred, and pH remained stable. Drug stability was maintained for 24 hours. Pharmacokinetic simulations showed that ketamine and lidocaine were within their minimum analgesic effect concentrations. Our results show that this three-drug mixture is chemically stable for up to 24 hours after mixing, with a pharmacokinetic simulation illustrating safe, clinically useful predicted plasma concentrations when using the described admixture.

Do habitat and elevation promote hybridization during secondary contact between three genetically distinct groups of warbling vireo (Vireo gilvus)?

Following postglacial expansion, secondary contact can occur between genetically distinct lineages. These genetic lineages may be associated with specific habitat or environmental variables and therefore, their distributions in secondary contact could reflect such conditions within these areas. Here we used mtDNA, microsatellite, and morphological data to study three genetically distinct groups of warbling vireo (Vireo gilvus) and investigate the role that elevation and habitat play in their distributions. We studied two main contact zones and within each contact zone, we examined two separate transects. Across the Great Plains contact zone, we found that hybridization between eastern and western groups occurs along a habitat and elevational gradient, whereas hybridization across the Rocky Mountain contact zone was not as closely associated with habitat or elevation. Hybrids in the Great Plains contact zone were more common in transitional areas between deciduous and mixed-wood forests, and at lower elevations (<1000 m). Hybridization patterns were similar along both Great Plains transects indicating that habitat and elevation play a role in hybridization between distinct eastern and western genetic groups. The observed patterns suggest adaptation to different habitats, perhaps originating during isolation in multiple Pleistocene refugia, is facilitating hybridization in areas where habitat types overlap.

A study of space charge induced non-linearity in the Single Line Of Sight camera.

A new generation of gated x-ray detectors at the National Ignition Facility has brought faster, enhanced imaging capabilities. Their performance is currently limited by the amount of signal they can be operated with before space charge effects in their electron tube start to compromise their temporal and spatial response. We present a technique to characterize this phenomenon and apply it to a prototype of such a system, the Single Line Of Sight camera. The results of this characterization are used to benchmark particle-in-cell simulations of the electrons drifting inside the detector, which are found to well reproduce the experimental data. These simulations are then employed to predict the optimum photon flux to the camera, with the goal to increase the quality of the images obtained on an experimental campaign while preventing the appearance of deleterious effects. They also offer some insights into some of the improvements that can be brought to the new pulse-dilation systems being built at Lawrence Livermore National Laboratory.

Timing characterization of fast hCMOS sensors.

We describe a method of analyzing gate profile data for ultrafast x-ray imagers that allows pixel-by-pixel determination of temporal sensitivity in the presence of substantial background oscillations. With this method, systematic timing errors in gate width and gate arrival time of up to 1 ns (in a 2 ns wide gate) can be removed. In-sensor variations in gate arrival and gate width are observed, with variations in each up to 0.5 ns. This method can be used to estimate the coarse timing of the sensor, even if errors up to several ns are present.

Effects of sodium salicylate and time postpartum on mammary tissue proliferation, gene transcript profile, and DNA methylation.

Previous studies have demonstrated nonsteroidal antiinflammatory drug treatment in early lactation had a positive impact on whole-lactation milk production in older cows. The objective of this study was to evaluate proliferative, transcriptional, and epigenetic changes in the mammary gland that could explain increased production responses due to nonsteroidal antiinflammatory drug treatment. Sodium salicylate (SAL; 125 g/d) or water (CON) were administered via oral drench to multiparous Holstein cows (n = 8/treatment) once daily for 3 d beginning approximately 24 h after parturition, and mammary tissue was collected on d 1, 4, and 45 postpartum. Day 1 tissue was collected immediately preceding the initial drench, and d 4 tissue was collected 24 h following the final drench. Blood was collected twice weekly and analyzed for plasma glucose, insulin, ß-hydroxybutyrate, free fatty acids, and prolactin. Cows were milked twice daily until d 7 of lactation, and thrice daily for the remainder of the study. Total RNA extracted from tissue was deep-sequenced and analyzed for differential gene expression using DESeq2. We detected no treatment effect on milk yield or plasma metabolites through 45 d of lactation; additionally, no change in mammary epithelial cell proliferation was detected when assessed by Ki67 labeling. Comparison of SAL versus CON revealed that only 16 of 18,286 genes were differentially expressed (false discovery rate <0.1) in mammary tissue collected on d 45, whereas no differentially expressed genes due to treatment were detected on d 1 or 4. Analysis of transcriptional differences over time showed downregulation of pathways related to immune cell recruitment and differentiation, and extensive overlap with pathways related to cholesterol synthesis and liver X receptor signaling. Global DNA methylation of mammary tissue was decreased for CON compared with SAL. Transcriptome analysis emphasized extensive involvement of immune-related signaling pathways in the switch from lactogenesis to galactopoiesis, and changes in methylation with SAL treatment merit future investigation into epigenetic effects on milk production.

Upgrade of the gated laser entrance hole imager G-LEH-2 on the National Ignition Facility.

A major upgrade has been implemented for the ns-gated laser entrance hole imager on the National Ignition Facility (NIF) to obtain high-quality data for Hohlraum physics study. In this upgrade, the single "Furi" hCMOS sensor (1024 × 448 pixel arrays with two-frame capability) is replaced with dual "Icarus" sensors (1024 × 512 pixel arrays with four-frame capability). Both types of sensors were developed by Sandia National Laboratories for high energy density physics experiments. With the new Icarus sensors, the new diagnostic provides twice the detection area with improved uniformity, wider temporal coverage, flexible timing setup, and greater sensitivity to soft x rays (<2 keV). These features, together with the fact that the diagnostic is radiation hardened and can be operated on the NIF for high neutron yield deuterium-triterium experiments, enable significantly greater return of data per experiment.

Oncolytic virus in gliomas: a review of human clinical investigations.

Gliomas remain one of the more frustrating targets for oncologic therapy. Glioma resistance to conventional therapeutics is a product of their immune-privileged milieu behind the blood-brain barrier, in addition to their suppressive effect on the immune response itself. Taking the lead from the growing success of immunotherapy for systemic cancers, such as lung cancer and melanoma, immunotherapeutics has emerged as a major player in the potential treatment of gliomas, with oncolytic viruses in particular showing significant promise as evidenced by the recent Breakthrough and Fast Tract Designations for PVSRIPO and DNX2401. This review serves as a useful and updated compendium of the completed human clinical investigations for several oncolytic viruses in the treatment of gliomas.

Behavior, health, and productivity of early-lactation dairy cows supplemented with molasses in automated milking systems.

The objective of this study was to determine the effect of molasses-based liquid feed (LF) supplementation within automated milking systems (AMS) on the behavior, health, and production of early-lactation dairy cows. In 6 commercial AMS dairy herds, 390 dairy cows were randomly assigned before calving to 1 of 2 treatments, within farm, balanced by parity: (1) control group (CON) receiving a standard AMS pellet [mean = 3.9 kg/d on a dry matter (DM) basis, n = 188], or (2) same amount of standard AMS pellet (mean = 3.6 kg/d on a DM basis, n = 202) plus 1 kg/d DM of LF for multiparous cows (1.6 kg/d as fed) and 0.88 kg/d DM for primiparous cows (1.4 kg/d as fed). Across farms, cows were fed partial mixed rations similar in ingredient and nutrient composition. Cows on the LF treatment received supplementation for the first 60 d in milk (DIM). Blood samples were taken 2 times/wk for the first 3 wk postcalving to assess ß-hydroxybutyrate (BHB). Samples with BHB ≥1.2 mmol/L were classified as a positive case of subclinical ketosis (SCK). Cow body condition and lameness were scored every 10 d postcalving until 60 DIM. No differences were detected between treatments for milk yield (average milk yield = 37.4 ± 1.98 kg/d) and milking frequency (average milking frequency = 3.2 ± 0.01 times/d). Similarly, no difference between treatments in daily AMS visits was detected (average AMS visits = 5.3 ± 0.3 times/d). However, CON cows experienced more AMS refusals/d than LF cows (LF = 1.6 refusals/d, CON = 2.1 refusals/d). No difference between treatments in daily rumination time was detected (average rumination time = 476 ± 10.8 min/d). Treatment was associated with the number of times cows tested positive for SCK; cows on LF had fewer repeated diagnoses of SCK, such that 15% of cows on LF had ≥3 diagnoses of SCK out of 5 tests, compared with 27% of CON cows. Cows receiving molasses tended to have lower blood BHB values at ∼15 DIM. Molasses supplementation also allowed cows to maintain a more stable body condition score, as LF cows tended to lose less body condition over the first 60 DIM compared with CON cows. Overall, the results of this study demonstrate that supplementing a molasses-based LF to AMS cows may help support the energy demands of milk production in early lactation and, thus, reduce the incidence of repeat positive SCK tests during that time period.

Macrophage-derived Wnt signaling increases endothelial permeability during skeletal muscle injury.

OBJECTIVE: The inflammatory response and the presence of macrophages are reported to be necessary for proper muscle regeneration. However, our understanding of the molecular mechanisms governing how macrophages signal to promote muscle regeneration is incomplete. METHODS AND RESULTS: Here we conditionally deleted Wls, which is required for Wnt secretion, from macrophages and examined the impact on endothelial permeability following muscle injury. The expression of Wnt ligands and Wls was increased in the tibialis anterior (TA) of mice 2 days following BaCl2 injury. Loss of macrophage Wls inhibited the loss of endothelial barrier function, as measured by transendothelial resistance and Evans blue dye permeability assays. Interestingly, the blockade in endothelial permeability correlated with reduced VEGF levels and pretreatment of wild type endothelial cells with a VEGFR2 blocking antibody was sufficient to reduce endothelial permeability induced by stimulated macrophage supernatant. We also found that macrophage Wls-null TAs had myocytes with reduced cross-sectional area 7 day post-injury suggesting a delay in muscle regeneration. CONCLUSION: Our results indicate that macrophage-derived Wnt signaling increases endothelial permeability in a VEGF-dependent fashion following muscle injury. Our findings implicate macrophages as a primary source of Wnt ligands following muscle injury and highlight the Wnt pathway as a therapeutic target following injury.

Effects of molasses-based liquid feed supplementation to a high-straw dry cow diet on feed intake, health, and performance of dairy cows across the transition period.

The objective of this study was to determine the effects of adding molasses-based liquid feed to a high-straw dry cow diet on intake, behavior, health, and performance of dairy cows across the transition period. Holstein cows (n = 40) entering their second lactation or greater were enrolled at dry-off (∼45 d before expected calving) and assigned to 1 of 2 treatments: a high-straw dry cow total mixed ration (TMR) with either (1) no molasses [CON; n = 20; 10.7% crude protein (CP), 1.45 Mcal/kg of net energy for lactation (NEL)] or (2) supplemented with 1 kg/cow daily [dry matter (DM) basis] of molasses-based liquid feed (LF; n = 20; 11.2% CP, 1.49 Mcal/kg of NEL). At calving all cows were fed the same lactating TMR (14.9% CP, 1.68 Mcal/kg of NEL) and followed for 28 d. During the dry period, cows fed the LF diet had greater DM intake (15.3 ± 0.24 vs. 13.4 ± 0.24 kg/d) and tended to have a shorter interval between meals (192.8 ± 8.38 vs. 216.5 ± 7.97 min) and more frequent meals (6.3 ± 0.27 vs. 5.6 ± 0.27 meals/d), while consuming their feed faster (0.09 ± 0.001 vs. 0.08 ± 0.001 kg of DM/min) compared with cows fed the CON diet. Regardless of treatment, cows sorted [100% × (actual intake/predicted intake)] against the longest ration particles (>19 mm), with cows fed the CON diet sorting more against these (81.1 ± 2.6 vs. 93.9 ± 2.6%). Cows fed the CON diet did not sort for or against short particles (<8, >4 mm; 100.2 ± 0.31%), but cows fed the LF diet tended to sort against these (99.2 ± 0.31%). Post-calving, cows fed the LF diet did not sort for long particles (98.5 ± 1.1%), but cows fed the CON diet continued to sort against these (96.3 ± 1.1%). Cows fed the LF diet had higher mean reticulorumen pH in the dry period (6.4 ± 0.05 vs. 6.2 ± 0.06 pH), in the first week after calving (CON = 5.7 ± 0.06 pH, LF = 5.8 ± 0.06 pH), and tended to have higher mean reticulorumen pH in the second week after calving (CON = 5.7 ± 0.06 pH, LF = 5.9 ± 0.06 pH). The results suggest that supplementing a molasses-based liquid feed in high-straw dry cow diets may improve intake and consistency in nutrients consumed during the dry period and in early lactation, as well as possibly promoting better rumen health across the transition period.

Effects of wheat straw chop length in high-straw dry cow diets on intake, health, and performance of dairy cows across the transition period.

The study investigated the effect of chop length of wheat straw in a high-straw dry cow diet on the feeding behavior, health, and performance of Holstein dairy cows across the transition period. Forty cows were enrolled at dry off, approximately 45 d before expected calving, and assigned to 1 of 2 dietary treatments containing (on a dry matter basis) 36% corn silage, 35% dry cow pellet, and 29% wheat straw. Diets were formulated to target an energy consumption of 19 Mcal/d of net energy for lactation per cow. The diets differed in the chop length of wheat straw: (1) long straw chopped with a 10.16-cm screen (LDD; n = 20), or (2) short straw chopped with a 2.54-cm screen (SDD; n = 20). Dietary treatments were fed throughout the dry period. Feeding behavior, rumination time, and reticulorumen pH were monitored electronically, and feed samples were collected to determine feed sorting. Blood samples were taken weekly, and body condition scores and body weights were taken every 2 wk. Postcalving, all cows were fed the same lactating cow total mixed ration for 28 d; the same measurements were recorded with the addition of daily milk yield and twice weekly milk components. During the dry period, cows fed the SDD treatment consumed more dry matter (15.6 vs. 15.0 kg/d), had more consistent dry matter intake in the week leading up to calving, had a faster feeding rate, consumed 0.4 kg/meal more, and had 0.6 fewer meals per day. No differences were observed in rumination time or mean reticulorumen pH between treatments in both the dry and lactating period. Regardless of treatment during the dry period, cows sorted against the long particles (>19 mm) and in favor of the short particles (<8, >4 mm), but cows fed the LDD treatment sorted to a greater extent than cows fed the SDD treatment. Dietary treatment in the dry period did not influence feed sorting behavior postcalving. Cows fed the LDD treatment had a greater daily change (decline) in rumen pH in the first week postcalving and had a greater blood ß-hydroxybutyrate concentration in the third week postcalving. Overall, the results from this study indicate that feeding shorter chopped wheat straw in a high-straw dry cow diet improved intake, reduced sorting during the dry period, and resulted in greater metabolic health and rumen stability in early lactation.

Moisture content of high-straw dry cow diets affects intake, health, and performance of transition dairy cows.

The objective of this study was to determine the effect of adding water to a high-straw dry cow diet on feeding behavior, health, and performance of dairy cows. Holstein cows (n = 40) entering their second or greater lactation were enrolled at dry off (∼45 d before expected calving) and assigned to 1 of 2 dietary treatments, consisting of a high-straw dry cow total mixed ration [TMR; 36% wheat straw, 41% corn silage, 23% pellet, on a dry matter (DM) basis, formulated for 11.6% crude protein, 1.35 Mcal/kg net energy for lactation] with (1) no water (CON; n = 20; DM = 53.4%) or (2) water added to decrease the DM by ∼10 percentage points (wet diet, WD; n = 20; DM = 45.4%). Upon calving, all cows were fed the same lactating TMR (45.2% DM, 14.9% crude protein, 1.68 Mcal/kg net energy for lactation) and followed for 28 d. Dry matter intake, feeding behavior, and rumination activity were recorded automatically. Fresh TMR and orts samples were collected 2×/wk to determine differences in sorting. A particle separator was used to separate feed samples into 4 fractions: long (>19 mm), medium (<19, >8 mm), short (<8, >4 mm), and fine (<4 mm) particles. Feed sorting was calculated as actual intake of each particle fraction expressed as a % of predicted intake. Cows fed WD had greater DM intake during the dry period than cows fed CON (CON = 13.4 ± 0.24, WD = 14.2 ± 0.24 kg/d), but rumination did not differ (CON = 522.2 ± 8.48, WD = 518.5 ± 8.69 min/d) between treatments. Cows fed WD tended to consume their feed faster (CON = 0.08 ± 0.001, WD = 0.09 ± 0.001 kg of DM/min) compared with CON cows. Regardless of treatment, cows sorted against the longest ration particles, with cows fed CON sorting more against these (CON = 81.1 ± 2.51%, WD = 94.9 ± 2.58%). Cows fed CON tended to sort more in favor of the medium particles than those fed WD (CON = 105.0 ± 0.36%, WD = 104.2 ± 0.37%). Postcalving DM intake and feeding behavior were not affected by dry diet treatment. Cows fed WD experienced a less rapid daily decline in rumen pH during the first 7 d after calving compared with those fed the CON diet. The results suggest that increasing the moisture content of high-straw dry cow diets, by adding water, may improve intake and reduce sorting against the longest ration particles, which may help promote consistency in targeted nutrients consumed during the dry period and greater rumen health in early lactation.

A retrospective analysis of the use of tap water iontophoresis for focal hyperhidrosis at a district general hospital: the patients' perspective.

Objective: Tap water iontophoresis as a treatment for focal hyperhidrosis is given as an initial series of treatments in hospital, followed by home maintenance treatments. Our study assessed quality of life and perception of hyperhidrosis with the use of iontophoresis. Materials and methods: All patients treated with iontophoresis at our hospital from 2012 to 2017 were retrospectively assessed (n = 82, mean age 34 years; 60% female). Fifty of the 82 patients (mean age 34 years; 60% female) had a pre-treatment DLQI (mean 12.6). Twenty-three of these patients (mean age 33 years; 60% female) had a paired pre- and post-treatment DLQI recorded. The average DLQI pre-treatment was 14.1 and post-treatment was 2.2. Therefore, the average reduction (improvement) was 11.9 (p < .05). Results: Thirty-eight of the 82 patients (46%) completed a telephone interview (mean age 35 years; 65% female). Of this cohort, 24 had an improvement in HDSS following treatment, and the remaining 14 patients had no change. Nine patients (24%) bought their own iontophoresis machine. In these patients, there was a higher average improvement in HDSS (1.8), compared to the total interviewed cohort (1.0). Conclusion: In conclusion, tap water iontophoresis can result in a significant improvement in perceived severity of hyperhidrosis and quality of life.