Evaluating H295R steroidogenesis assay data for robust interpretation.

The in vitro H295R steroidogenesis assay (OECD TG 456) is used to determine a chemical's potential to interfere with steroid hormone synthesis/metabolism. As positive outcomes in this assay can trigger significant higher tiered testing, we compiled a stakeholder database of reference and test item H295R data to characterize assay outcomes. Information concerning whether a Level 5 reproductive toxicity study was triggered due to a positive outcome in the H295R assay was also included. Quality control acceptance criteria were not always achieved, suggesting this assay is challenging to conduct within the guideline specifications. Analysis of test item data demonstrated that pairwise significance testing to controls allowed for overly sensitive statistically significant positive outcomes, which likely contribute to the assay's high positive hit rate. Complementary interpretation criteria (e.g., 1.5-fold change threshold) markedly reduced the rate of equivocal and positive outcomes thus improving identification of robust positive effects in the assay. Finally, a case study (positive H295R outcome and no endocrine adversity in vivo) is presented, which suggests that stricter data interpretation criteria could refine necessary in vivo follow-up testing. Overall, the described additional criteria could improve H295R data interpretation and help inform on how to best leverage this assay for regulatory purposes.

Compact thermal imager: a flight demonstration of infrared technology for Earth observations.

During 2019, an infrared camera, the compact thermal imager (CTI), recorded 15 million images of the Earth from the International Space Station. CTI is based on strained-layer superlattice (SLS) detector technology. The camera covered the spectral range from 3 to 11 µm in two spectral channels, 3.3-5.4 and 7.8-10.7 µm. Individual image frames were 26×21km2 projected on the ground, with 82 m pixel resolution. A frame time of 2.54 s created continuous image swaths with a 13% along-track image overlap. Upper limits determined on the ground and in flight for the electronic offset, read noise, and dark current demonstrated the stability of the SLS detector and camera over many months. Temperature calibration was established using a combination of preflight and in-flight measurements. A narrowband approximation of temperature as a function of photon counts produced an analytic relationship covering a temperature range of 0°-400°C. Examples of CTI images illustrate temperature retrievals over sea ice, urban and agricultural areas, desert, and wildfires.

Phosphorylation of intracellular signalling molecules in peripheral blood cells from patients with psoriasis on originator or biosimilar infliximab.

BACKGROUND: Psoriasis vulgaris is a chronic, inflammatory skin disease characterized by a dysregulated immune response and it is associated with substantial systemic comorbidities. Biological drugs such as tumour necrosis factor (TNF)-α inhibitors can ameliorate the disease but are expensive. Biosimilar drugs have the same amino-acid sequence as the originator, but differences in manufacturing can affect biological activity, efficacy and tolerability. OBJECTIVES: To explore potential differences in intracellular phosphorylation of signalling molecules in peripheral blood cells from patients with psoriasis treated with the TNF-α inhibitor infliximab compared with healthy controls, and to investigate if the phosphorylation pattern was influenced by switching from the originator infliximab to the biosimilar CT-P13. METHODS: By flow cytometry, we measured phosphorylation of nuclear factor kappa B, extracellular signal-regulated kinase 1/2, p38 mitogen-activated protein kinase and signal transducer and activator of transcription 3, before and after TNF-α stimulation in monocytes and T, B, natural killer and CD3+  CD56+ cells from 25 patients with psoriasis treated with infliximab and 19 healthy controls. RESULTS: At inclusion, phosphorylation levels of peripheral blood mononuclear cells (PBMCs) were increased in patients with psoriasis compared with healthy controls, even though clinical remission had already been achieved. Phosphorylation levels declined in patients on both originator infliximab and biosimilar during continued treatment. No significant differences were detected between the two medications after 12 months. CONCLUSIONS: Patients with psoriasis on infliximab have higher activation levels of PBMCs than do healthy controls, possibly reflecting systemic inflammation. Switching from the originator infliximab to biosimilar CT-P13 did not affect phosphorylation levels or clinical parameters, suggesting that CT-P13 is a noninferior treatment alternative to the originator infliximab.

Route-learning strategies in typical and atypical development; eye tracking reveals atypical landmark selection in Williams syndrome.

BACKGROUND: Successful navigation is crucial to everyday life. Individuals with Williams syndrome (WS) have impaired spatial abilities. This includes a deficit in spatial navigation abilities such as learning the route from A to B. To-date, to determine whether participants attend to landmarks when learning a route, landmark recall tasks have been employed after the route learning experience. Here, we combined virtual reality and eye tracking technologies, for the first time, to measure landmark use in typically developing (TD) children and participants with WS during route-learning. METHOD: Nineteen individuals with WS were asked to learn a route in a sparse environment (few landmarks) and in a rich environment (many landmarks) whilst their eye movements were recorded. Looking times towards landmarks were compared to TD children aged 6, 8 and 10 years. Changes in attention to landmarks during the learning process were also recorded. RESULTS: The WS group made fewer looks to landmarks overall, but all participants looked for longer at landmarks that were at junctions and along the paths of the maze than landmarks that were in the distance. Few differences were observed in route learning between the sparse and rich environments. In contrast to the TD groups, those in the WS group were as likely to look at non-unique landmarks as landmarks at junctions and on paths. DISCUSSION: The current results demonstrate that attention to landmarks during route learning reflects the types of landmarks remembered in memory tasks, that individuals with WS can learn a route if given sufficient exposure, but that this is accomplished within the context of an impaired ability to select appropriate landmarks.

Controlled infection with a therapeutic virus defines the activation kinetics of human natural killer cells in vivo.

Human natural killer (NK) cells play an important role in anti-viral immunity. However, studying their activation kinetics during infection is highly problematic. A clinical trial of a therapeutic virus provided an opportunity to study human NK cell activation in vivo in a controlled manner. Ten colorectal cancer patients with liver metastases received between one and five doses of oncolytic reovirus prior to surgical resection of their tumour. NK cell surface expression of the interferon-inducible molecules CD69 and tetherin peaked 24-48 h post-infection, coincident with a peak of interferon-induced gene expression. The interferon response and NK cell activation were transient, declining by 96 h post-infection. Furthermore, neither NK cell activation nor the interferon response were sustained in patients undergoing multiple rounds of virus treatment. These results show that reovirus modulates human NK cell activity in vivo and suggest that this may contribute to any therapeutic effect of this oncolytic virus. Detection of a single, transient peak of activation, despite multiple treatment rounds, has implications for the design of reovirus-based therapy. Furthermore, our results suggest the existence of a post-infection refractory period when the interferon response and NK cell activation are blunted. This refractory period has been observed previously in animal models and may underlie the enhanced susceptibility to secondary infections that is seen following viral infection.

Quantitative diet analysis of four mesopredators from a coral reef.

The diets of four common mesopredator fishes were examined in the back-reef habitat of a subtropical fringing reef system during the summer months. Quantitative gut content analyses revealed that crustaceans, represented >60% of ingested prey (% mass) by the latticed sand-perch Parapercis clathrata, brown dottyback Pseudochromis fuscus and half-moon grouper Epinephelus rivulatus. Dietary analyses also provided insights into ontogenetic shifts. Juvenile P. fuscus ingested large numbers of crustaceans (amphipods and isopods); these small prey were rarely found in larger individuals (<1% of ingested mass). Fishes also made an important contribution to the diets of all three species representing 10-30% of ingested mass. Conversely, the sand lizardfish Synodus dermatogenys fed exclusively on fishes including clupeids, gobies and labrids. Differences in the gut contents of the four species recorded were not apparent using stable isotope analysis of muscle tissues. The similarity of δ(13) C values in muscle tissues suggested that carbon within prey was derived from primary producers, with comparable carbon isotope signatures to corals and macroalgae, whilst similarities in δ(15) N values indicated that all four species belonged to the same trophic level. Thus, interspecific differences between mesopredator diets were undetectable when using stable isotope analysis which suggests that detailed elucidation of trophic pathways requires gut content analyses.

Critical research needs for managing coral reef marine protected areas: perspectives of academics and managers.

Marine protected areas (MPAs) are a primary policy instrument for managing and protecting coral reefs. Successful MPAs ultimately depend on knowledge-based decision making, where scientific research is integrated into management actions. Fourteen coral reef MPA managers and sixteen academics from eleven research, state and federal government institutions each outlined at least five pertinent research needs for improving the management of MPAs situated in Australian coral reefs. From this list of 173 key questions, we asked members of each group to rank questions in order of urgency, redundancy and importance, which allowed us to explore the extent of perceptional mismatch and overlap among the two groups. Our results suggest the mismatch among MPA managers and academics is small, with no significant difference among the groups in terms of their respective research interests, or the type of questions they pose. However, managers prioritised spatial management and monitoring as research themes, whilst academics identified climate change, resilience, spatial management, fishing and connectivity as the most important topics. Ranking of the posed questions by the two groups was also similar, although managers were less confident about the achievability of the posed research questions and whether questions represented a knowledge gap. We conclude that improved collaboration and knowledge transfer among management and academic groups can be used to achieve similar objectives and enhance the knowledge-based management of MPAs.

Drivers of variation in occurrence, abundance, and behaviour of sharks on coral reefs.

Quantifying the drivers of population size in reef sharks is critical for the development of appropriate conservation strategies. In north-west Australia, shark populations inhabit coral reefs that border growing centres of human population, industry, and tourism. However, we lack baseline data on reef sharks at large spatial scales (hundreds of km) that might enable managers to assess the status of shark populations in the face of future development in this region. Here, we examined the occurrence, abundance and behaviour of apex (Galeocerdo cuvier, Carcharhinus plumbeus) and reef (C. amblyrhynchos, C. melanopterus, Triaenodon obesus) sharks using > 1200 deployments of baited remote underwater stereo-video systems (stereo-BRUVs) across > 500 km of coastline. We found evidence for species-specific influences of habitat and fishing activities on the occurrence (probability of observation), abundance (MaxN) and behaviour of sharks (time of arrival to the stereo-BRUVs and likelihood of feeding). Although the presence of management zoning (No-take areas) made little difference to most species, C. amblyrhynchos were more common further from boat ramps (a proxy of recreational fishing pressure). Time of arrival for all species was also influenced by distance to boat ramp, although patterns varied among species. Our results demonstrate the capacity for behavioural metrics to complement existing measures of occurrence and abundance in assessing the potential impact of human activities on shark populations.

Gravity Probe B: final results of a space experiment to test general relativity.

Gravity Probe B, launched 20 April 2004, is a space experiment testing two fundamental predictions of Einstein's theory of general relativity (GR), the geodetic and frame-dragging effects, by means of cryogenic gyroscopes in Earth orbit. Data collection started 28 August 2004 and ended 14 August 2005. Analysis of the data from all four gyroscopes results in a geodetic drift rate of -6601.8±18.3 mas/yr and a frame-dragging drift rate of -37.2±7.2 mas/yr, to be compared with the GR predictions of -6606.1 mas/yr and -39.2 mas/yr, respectively ("mas" is milliarcsecond; 1 mas=4.848×10(-9) rad).

Development of a novel micro-bead force spectroscopy approach to measure the ability of a thermo-active polymer to remove bacteria from a corneal model.

Microbial keratitis occurs from the infection of the cornea by fungi and or bacteria. It remains one of the most common global causes of irreversible blindness accounting for 3.5% (36 million) of blind people as of 2015. This paper looks at the use of a bacteria binding polymer designed to bind Staphylococcus aureus and remove it from the corneal surface. Mechanical unbinding measurements were used to probe the interactions of a thermo-active bacteria-binding polymer, highly-branched poly(N-isopropyl acrylamide), functionalised with modified vancomycin end groups (HB-PNIPAM-Van) to bacteria placed on rabbit corneal surfaces studied ex-vivo. This was conducted during sequential temperature phase transitions of HB-PNIPAM-Van-S. aureus below, above and below the lower critical solution temperature (LCST) in 3 stages, in-vitro, using a novel micro-bead force spectroscopy (MBFS) approach via atomic force microscopy (AFM). The effect of temperature on the functionality of HB-PNIPAM-Van-S. aureus showed that the polymer-bacteria complex reduced the work done in removing bacterial aggregates at T > LCST (p < 0.05), exhibiting reversibility at T < LCST (p < 0.05). At T < LCST, the breaking force, number of unbinding events, percentage fitted segments in the short and long range, and the percentage of unbinding events occurring in the long range (> 2.5 µm) increased (p < 0.05). Furthermore, the LCST phase transition temperature showed 100 × more unbinding events in the long-range z-length (> 2.5 µm) compared to S. aureus aggregates only. Here, we present the first study using AFM to assess the reversible mechanical impact of a thermo-active polymer-binding bacteria on a natural corneal surface.

Dataset of 18O and 2H in streamflow across Canada: A national resource for tracing water sources, water balance and predictive modelling.

Oxygen-18 and deuterium were measured in streamflow samples collected from 331 gauging stations across Canada during 2013 to 2019. This dataset includes 9206 isotopic analyses made on 4603 individual water samples, and an additional 1259 analysis repeats for quality assurance/quality control. We also include arithmetic and flow-weighted averages, and other basic statistics for stations where adequate data were available. Station data are provided including station code, name, province, latitude, longitude and drainage area. Flow data were extracted from the historical database of the Water Survey of Canada. Details on the preliminary application of these data are provided in "18O and 2H in streamflow across Canada" [1]. Overall, these data are expected to be useful when combined with precipitation datasets and analytical or numerical models for water resource management and planning, including tracing streamflow source, water balance, evapotranspiration partitioning, residence time analysis, and early detection of climate and land use changes in Canada.

Association of Src tyrosine kinase with a human potassium channel mediated by SH3 domain.

The human Kv1.5 potassium channel (hKv1.5) contains proline-rich sequences identical to those that bind to Src homology 3 (SH3) domains. Direct association of the Src tyrosine kinase with cloned hKv1.5 and native hKv1.5 in human myocardium was observed. This interaction was mediated by the proline-rich motif of hKv1.5 and the SH3 domain of Src. Furthermore, hKv1.5 was tyrosine phosphorylated, and the channel current was suppressed, in cells coexpressing v-Src. These results provide direct biochemical evidence for a signaling complex composed of a potassium channel and a protein tyrosine kinase.

Ultrastructural studies of Phellinus sulphurascens infection of Douglas-fir roots and immunolocalization of host pathogenesis-related proteins.

Interactions between roots of Douglas-fir (DF; Pseudotsuga menziesii) seedlings and the laminated root rot fungus Phellinus sulphurascens were investigated using scanning and transmission electron microscopy and immunogold labelling techniques. Scanning electron micrographs revealed that P. sulphurascens hyphae colonize root surfaces and initiate the penetration of root epidermal tissues by developing appressoria within 2 d postinoculation (dpi). During early colonization, intra- and intercellular fungal hyphae were detected. They efficiently disintegrate cellular components of the host including cell walls and membranes. P. sulphurascens hyphae penetrate host cell walls by forming narrow hyphal tips and a variety of haustoria-like structures which may play important roles in pathogenic interactions. Ovomucoid-WGA (wheat germ agglutinin) conjugated gold particles (10 nm) confirmed the occurrence and location of P. sulphurascens hyphae, while four specific host pathogenesis-related (PR) protein antibodies conjugated with protein A-gold complex (20 nm) showed the localization and abundance of these PR proteins in infected root tissues. A thaumatin-like protein and an endochitinase-like protein were both strongly evident and localized in host cell membranes. A DF-PR10 protein was localized in the cell walls and cytoplasm of host cells while an antimicrobial peptide occurred in host cell walls. A close association of some PR proteins with P. sulphurascens hyphae suggests their potential antifungal activities in DF roots.

The role of glycogen synthase kinase-3beta in normal haematopoiesis, angiogenesis and leukaemia.

Glycogen synthase kinase 3 beta (GSK-3beta) was one of the first kinases identified and studied, initially for its role in the regulation of glycogen synthesis. Over the past decade, interest in GSK-3beta has grown far beyond glycogen metabolism, and this is due in large measure to the critical role that GSK-3beta plays in the regulation of many other cellular processes, particularly cell proliferation and apoptosis. GSK-3beta has been shown to regulate the proteolysis and sub-cellular compartmentalization of a number of proteins directly involved in the regulation of cell cycling, proliferation, differentiation and apoptosis. GSK-3beta also regulates the degradation of proteins that regulate gene expression and thus affects a variety of important cell functions. Specifically, GSK-3beta controls the degradation of beta-catenin, the main effector of Wnt that regulates haematopoiesis and stem cell function. In this case GSK-3beta is a negative regulator of Wnt. In contrast, GSK-3beta positively regulates NF-kappaB, another important biochemical pathway also involved in the regulation of multiple aspects of normal and aberrant haematopoiesis. GSK-3beta regulates degradation of IkappaB, a central inhibitor of NF-kappaB. In this way, GSK-3beta acts to control the resistance of leukaemic cells to chemotherapy through the modulation of NF-kappaB, a critical factor in maintaining leukaemic cell growth. In addition, GSK-3beta regulates the pro-inflammatory activity of NF-kappaB. As GSK-3beta is a pleiotropic regulator, inhibitors may increase the range of novel anti-leukaemic and anti-inflammatory drugs that control immune response.

Lung deposition of droplet aerosols in monkeys.

Nonhuman primates are often the animal models of choice to study the infectivity and therapy of inhaled infectious agents. Most animal models for inhaled infectious diseases use aerosol/droplets generated by an atomization technique such as a Collison nebulizer that produces particles in the size range of 1 to 3 microm in diameter. There are few data in the literature on deposition patterns in monkeys. Our study was designed to measure the deposition pattern in monkeys using droplets having diameters of 2 and 5 microm using an exposure system designed to expose monkeys to aerosols of infectious agents. Six cynomolgus monkeys were exposed to droplets. The aerosol solution was generated from a Vero cell supernate containing DMEM + 10% fetal bovine serum tagged with Tc-99m radiolabel. Collison and Retec nebulizers were used to generate small and large droplets, respectively. The particle size (as determined from a cascade impactor) showed an activity median aerodynamic diameter (AMAD) of 2.3 and 5.1 microm for the Collison and Retec nebulizer, respectively. The animals were anesthetized, placed in a plethysmography box, and exposed to the aerosol. The deposition pattern was determined using a gamma camera. Deposition in the head airways was 39% and 58% for 2.3- and 5.1-microm particle aerosols, respectively, whereas the deposition in the deep lung was 12% and 8%, respectively. This information will be useful in developing animal models for inhaled infectious agents.

Cancer incidence in a religious isolate of Alberta, Canada, 1953-74.

Members of a religious isolate who live in approximately 240 farming colonies in the Canadian prairie provinces and the United States border states were studied. The sect's 6,700 members living in the province of Alberta, Canada, comprise more than 30% of this sect's population of North America. The numbers of their cancer cases ascertained from 1953 to 1974 in Alberta were compared to those expected from Alberta Cancer Registry rates. The overall incidence of registered cases of cancer among the religious isolate's females was significantly less than expected (48 observed, 74.2 expected), and in the males the overall incidence of cancer did not differ from that expected (52 observed, 56.5 expected). Significantly fewer cases of lung cancer than expected were found in males, and significantly fewer cases of carcinoma in situ of the cervix uteri were found in females. Finally, significantly higher incidence of stomach cancer was found in the sect's males. Data on a family with two cases of stomach cancer contributed to this observed excess of stomach cancer.

Modulation of the Kv1.3 potassium channel by receptor tyrosine kinases.

The voltage-dependent potassium channel, Kv1.3, is modulated by the epidermal growth factor receptor (EGFr) and the insulin receptor tyrosine kinases. When the EGFr and Kv1.3 are coexpressed in HEK 293 cells, acute treatment of the cells with EGF during a patch recording can suppress the Kv1.3 current within tens of minutes. This effect appears to be due to tyrosine phosphorylation of the channel, as it is blocked by treatment with the tyrosine kinase inhibitor erbstatin, or by mutation of the tyrosine at channel amino acid position 479 to phenylalanine. Previous work has shown that there is a large increase in the tyrosine phosphorylation of Kv1.3 when it is coexpressed with the EGFr. Pretreatment of EGFr and Kv1.3 cotransfected cells with EGF before patch recording also results in a decrease in peak Kv1.3 current. Furthermore, pretreatment of cotransfected cells with an antibody to the EGFr ligand binding domain (alpha-EGFr), which blocks receptor dimerization and tyrosine kinase activation, blocks the EGFr-mediated suppression of Kv1.3 current. Insulin treatment during patch recording also causes an inhibition of Kv1.3 current after tens of minutes, while pretreatment for 18 h produces almost total suppression of current. In addition to depressing peak Kv1.3 current, EGF treatment produces a speeding of C-type inactivation, while pretreatment with the alpha-EGFr slows C-type inactivation. In contrast, insulin does not influence C-type inactivation kinetics. Mutational analysis indicates that the EGF-induced modulation of the inactivation rate occurs by a mechanism different from that of the EGF-induced decrease in peak current. Thus, receptor tyrosine kinases differentially modulate the current magnitude and kinetics of a voltage-dependent potassium channel.

Life events and prisoners.

This study explores retrospectively the relationship of the accumulation of life events as it relates to prison incarceration and extends further the concept that coping with increasing environmental changes results in a variety of overt behaviors. The prison sample comprised 176 male inmates of a federal prison (McNeil Island, Washington) and a state penitentiary (Walla Walla, Washington). Life change scores were derived from the Schedule of Recent Experience (SRE). There was an escalation of annual life change scores of prisoners, indicating the mounting frequency of occurrence of life events prior to incarceration. The SRE may have value in the prediction of socially deviant behavior as with health changes. Variables seen as influencing life change scores were race, age, and education. Analyses of life event frequencies as compared to a normative group indicated that prisoners have evolved a coping life-style that reflects antisocial and criminal behavior.