Preliminary experimental results for a cryogenic brush seal configuration

Preliminary fluid nitrogen flow data are reported for a five-brush, ceramic-coated-rub-runner brush seal system, where the brushes and the rub runner were placed at each end of a centrally pressurized multifunction tester ('back-to-back' set of brushes) and tested at rotor speeds of 0, 10, 18, and 22.5 krpm. After testing, both the brushes and the ceramic-coated rub runner appeared pristine. The coating withstood both the thermomechanical and dynamic loadings with minor wear track scarring. The bristle tips showed some indication of material shearing (smearing) wear. The Ergun porous flow equation was applied to the brush seal data. The Ergun relation, which required heuristic information to characterize the coefficients, fit the gaseous data but was in poor agreement with the fluid results. The brush seal exit conditions were two phase. Two-phase, choked-flow design charts were applied but required one data point at each rotor speed to define the (C(sub f)A x Constant) flow and area coefficients. Reasonable agreement between prediction and data was found, as expected, but such methods are not to be construed as two-phase-flow brush seal analyses

Small scale heater tests in argillite of the Eleana Formation at the Nevada Test Site

Near-surface heater tests were run in the Eleana Formation at the Nevada Test Site, in an effort to evaluate argillaceous rock for nuclear waste storage. The main test, which employed a full-scale heater with a thermal output approximating commercial borosilicate waste, was designed to operate for several months. Two smaller, scaled tests were run prior to the full-scale test. This report develops the thermal scaling laws, describes the pretest thermal and thermomechanical analysis conducted for these two tests, and discusses the material properties data used in the analyses. In the first test, scaled to a large heater of 3.5 kW power, computed heater temperatures were within 7% of measured values for the entire 96-hour test run. The second test, scaled to a large heater having 5.0 kW power, experienced periodic water in-flow onto the heater, which tended to damp the temperature. For the second test, the computed temperatures were within 7% of measured for the first 20 hours. After this time, the water effect became significant and the measured temperatures were 15 to 20% below those predicted. On the second test, rock surface spallation was noted in the bore hole above the heater, as predicted. The scaled tests indicated that in-situ argillite would not undergo major thermostructural failure during the follow-on, 3.5 kW, full-scale test. 24 figures, 6 tables

Hypoxia and hyperglycaemia determine why some endometrial tumours fail to respond to metformin

High expression of Ki67, a proliferation marker, is associated with reduced endometrial cancer-specific survival. Pre-surgical metformin reduces tumour Ki-67 expression in some women with endometrial cancer. Metformin's anti-cancer activity may relate to effects on cellular energy metabolism. Since tumour hypoxia and glucose availability are major cellular redox determinants, we evaluated their role in endometrial cancer response to metformin. Endometrial cancer biopsies from women treated with pre-surgical metformin were tested for the hypoxia markers, HIF-1α and CA-9. Endometrial cancer cell lines were treated with metformin in variable glucose concentrations in normoxia or hypoxia and cell viability, mitochondrial biogenesis, function and energy metabolism were assessed. In women treated with metformin (n = 28), Ki-67 response was lower in hypoxic tumours. Metformin showed minimal cytostatic effects towards Ishikawa and HEC1A cells in conventional medium (25 mM glucose). In low glucose (5.5 mM), a dose-dependent cytostatic effect was observed in normoxia but attenuated in hypoxia. Tumours treated with metformin showed increased mitochondrial mass (n = 25), while in cultured cells metformin decreased mitochondrial function. Metformin targets mitochondrial respiration, however, in hypoxic, high glucose conditions, there was a switch to glycolytic metabolism and decreased metformin response. Understanding the metabolic adaptations of endometrial tumours may identify patients likely to derive clinical benefit from metformin

Eleana near-surface heater experiment final report

This report summarizes the results of a near-surface heater experiment operated at a depth of 23 m in argillite within the Eleana Formation on the Nevada Test Site (NTS). The test geometrically simulated emplacement of a single canister of High-Level Waste (HLW) and was operated at a power level of 2.5 kW for 21 days, followed by 3.8 kW to 250 days, when the power was turned off. Below 85 to 100{sup 0}C, there was good agreement between modeled and measured thermal results in the rock and in the emplacement hole, except for transient transport of water in the heater hole. Above 100{sup 0}C, modeled and measured thermal results increasingly diverged, indicating that the in-situ rock-mass thermal conductivity decreased as a result of dehydration more than expected on the basis of matrix properties. Correlation of thermomechanical modeling and field results suggests that this decrease was caused by strong coupling of thermal and mechanical behavior of the argillite at elevated temperatures. No hole-wall decrepitation was observed in the experiment; this fact and the codrrelation of modeled and measured results at lower temperatures indicate that there is no a priori reason to eliminate argillaceous rocks from further consideration as a host rock for nuclear wastes

Small scale heater tests in argillite of the Eleana Formation at the Nevada Test Site

Near-surface heater tests were run in the Eleana Formation at the Nevada Test Site, in an effort to evaluate argillaceous rock for nuclear waste storage. The main test, which employed a full-scale heater with a thermal output approximating commercial borosilicate waste, was designed to operate for several months. Two smaller, scaled tests were run prior to the full-scale test. This report develops the thermal scaling laws, describes the pretest thermal and thermomechanical analysis conducted for these two tests, and discusses the material properties data used in the analyses. In the first test, scaled to a large heater of 3.5 kW power, computed heater temperatures were within 7% of measured values for the entire 96-hour test run. The second test, scaled to a large heater having 5.0 kW power, experienced periodic water in-flow onto the heater, which tended to damp the temperature. For the second test, the computed temperatures were within 7% of measured for the first 20 hours. After this time, the water effect became significant and the measured temperatures were 15 to 20% below those predicted. On the second test, rock surface spallation was noted in the bore hole above the heater, as predicted. The scaled tests indicated that in-situ argillite would not undergo major thermostructural failure during the follow-on, 3.5 kW, full-scale test. 24 figures, 6 tables

An online breathing and wellbeing programme (ENO Breathe) for people with persistent symptoms following COVID-19: a parallel-group, single-blind, randomised controlled trial

BACKGROUND: There are few evidence-based interventions for long COVID; however, holistic approaches supporting recovery are advocated. We assessed whether an online breathing and wellbeing programme improves health related quality-of-life (HRQoL) in people with persisting breathlessness following COVID-19. METHODS: We conducted a parallel-group, single-blind, randomised controlled trial in patients who had been referred from one of 51 UK-based collaborating long COVID clinics. Eligible participants were aged 18 years or older; were recovering from COVID-19 with ongoing breathlessness, with or without anxiety, at least 4 weeks after symptom onset; had internet access with an appropriate device; and were deemed clinically suitable for participation by one of the collaborating COVID-19 clinics. Following clinical assessment, potential participants were given a unique online portal code. Participants were randomly assigned (1:1) to either immediate participation in the English National Opera (ENO) Breathe programme or to usual care. Randomisation was done by the research team using computer-generated block randomisation lists, with block size 10. The researcher responsible for randomisation was masked to responses. Participants in the ENO Breathe group participated in a 6-week online breathing and wellbeing programme, developed for people with long COVID experiencing breathlessness, focusing on breathing retraining using singing techniques. Those in the deferred group received usual care until they exited the trial. The primary outcome, assessed in the intention-to-treat population, was change in HRQoL, assessed using the RAND 36-item short form survey instrument mental health composite (MHC) and physical health composite (PHC) scores. Secondary outcome measures were the chronic obstructive pulmonary disease assessment test score, visual analogue scales (VAS) for breathlessness, and scores on the dyspnoea-12, the generalised anxiety disorder 7-item scale, and the short form-6D. A thematic analysis exploring participant experience was also conducted using qualitative data from focus groups, survey responses, and email correspondence. This trial is registered with ClinicalTrials.gov, NCT04830033. FINDINGS: Between April 22 and May 25, 2021, 158 participants were recruited and randomly assigned. Of these, eight (5%) individuals were excluded and 150 participants were allocated to a treatment group (74 in the ENO Breathe group and 76 in the usual care group). Compared with usual care, ENO Breathe was associated with an improvement in MHC score (regression coefficient 2·42 [95% CI 0·03 to 4·80]; p=0·047), but not PHC score (0·60 [-1·33 to 2·52]; p=0·54). VAS for breathlessness (running) favoured ENO Breathe participation (-10·48 [-17·23 to -3·73]; p=0·0026). No other statistically significant between-group differences in secondary outcomes were observed. One minor self-limiting adverse event was reported by a participant in the ENO Breathe group who felt dizzy using a computer for extended periods. Thematic analysis of ENO Breathe participant experience identified three key themes: (1) improvements in symptoms; (2) feeling that the programme was complementary to standard care; and (3) the particular suitability of singing and music to address their needs. INTERPRETATION: Our findings suggest that an online breathing and wellbeing programme can improve the mental component of HRQoL and elements of breathlessness in people with persisting symptoms after COVID-19. Mind-body and music-based approaches, including practical, enjoyable, symptom-management techniques might have a role supporting recovery. FUNDING: Imperial College London

Drosophila DNA polymerase theta utilizes both helicase-like and polymerase domains during microhomology-mediated end joining and interstrand crosslink repair

Double strand breaks (DSBs) and interstrand crosslinks (ICLs) are toxic DNA lesions that can be repaired through multiple pathways, some of which involve shared proteins. One of these proteins, DNA Polymerase theta (Pol theta), coordinates a mutagenic DSB repair pathway named microhomology-mediated end joining (MMEJ) and is also a critical component for bypass or repair of ICLs in several organisms. Pol theta contains both polymerase and helicase-like domains that are tethered by an unstructured central region. While the role of the polymerase domain in promoting MMEJ has been studied extensively both in vitro and in vivo, a function for the helicase-like domain, which possesses DNA-dependent ATPase activity, remains unclear. Here, we utilize genetic and biochemical analyses to examine the roles of the helicase-like and polymerase domains of Drosophila Pol theta. We demonstrate an absolute requirement for both polymerase and ATPase activities during ICL repair in vivo. However, similar to mammalian systems, polymerase activity, but not ATPase activity, is required for ionizing radiation-induced DSB repair. Using a site-specific break repair assay, we show that overall end-joining efficiency is not affected in ATPase-dead mutants, but there is a significant decrease in templated insertion events. In vitro, Pol theta can efficiently bypass a model unhooked nitrogen mustard crosslink and promote DNA synthesis following microhomology annealing, although ATPase activity is not required for these functions. Together, our data illustrate the functional importance of the helicase-like domain of Pol theta and suggest that its tethering to the polymerase domain is important for its multiple functions in DNA repair and damage tolerance

DNA resection in eukaryotes: deciding how to fix the break

DNA double-strand breaks are repaired by different mechanisms, including homologous recombination and nonhomologous end-joining. DNA-end resection, the first step in recombination, is a key step that contributes to the choice of DSB repair. Resection, an evolutionarily conserved process that generates single-stranded DNA, is linked to checkpoint activation and is critical for survival. Failure to regulate and execute this process results in defective recombination and can contribute to human disease. Here, I review recent findings on the mechanisms of resection in eukaryotes, from yeast to vertebrates, provide insights into the regulatory strategies that control it, and highlight the consequences of both its impairment and its deregulation

State-of-the-art of 3D cultures (organs-on-a-chip) in safety testing and pathophysiology.

Integrated approaches using different in vitro methods in combination with bioinformatics can (i) increase the success rate and speed of drug development; (ii) improve the accuracy of toxicological risk assessment; and (iii) increase our understanding of disease. Three-dimensional (3D) cell culture models are important building blocks of this strategy which has emerged during the last years. The majority of these models are organotypic, i.e., they aim to reproduce major functions of an organ or organ system. This implies in many cases that more than one cell type forms the 3D structure, and often matrix elements play an important role. This review summarizes the state of the art concerning commonalities of the different models. For instance, the theory of mass transport/metabolite exchange in 3D systems and the special analytical requirements for test endpoints in organotypic cultures are discussed in detail. In the next part, 3D model systems for selected organs--liver, lung, skin, brain--are presented and characterized in dedicated chapters. Also, 3D approaches to the modeling of tumors are presented and discussed. All chapters give a historical background, illustrate the large variety of approaches, and highlight up- and downsides as well as specific requirements. Moreover, they refer to the application in disease modeling, drug discovery and safety assessment. Finally, consensus recommendations indicate a roadmap for the successful implementation of 3D models in routine screening. It is expected that the use of such models will accelerate progress by reducing error rates and wrong predictions from compound testing

Competition between Replicative and Translesion Polymerases during Homologous Recombination Repair in Drosophila

In metazoans, the mechanism by which DNA is synthesized during homologous recombination repair of double-strand breaks is poorly understood. Specifically, the identities of the polymerase(s) that carry out repair synthesis and how they are recruited to repair sites are unclear. Here, we have investigated the roles of several different polymerases during homologous recombination repair in Drosophila melanogaster. Using a gap repair assay, we found that homologous recombination is impaired in Drosophila lacking DNA polymerase zeta and, to a lesser extent, polymerase eta. In addition, the Pol32 protein, part of the polymerase delta complex, is needed for repair requiring extensive synthesis. Loss of Rev1, which interacts with multiple translesion polymerases, results in increased synthesis during gap repair. Together, our findings support a model in which translesion polymerases and the polymerase delta complex compete during homologous recombination repair. In addition, they establish Rev1 as a crucial factor that regulates the extent of repair synthesis