Remote ischaemic conditioning: defining critical criteria for success-report from the 11th Hatter Cardiovascular Workshop.

The Hatter Cardiovascular Institute biennial workshop, originally scheduled for April 2020 but postponed for 2 years due to the Covid pandemic, was organised to debate and discuss the future of Remote Ischaemic Conditioning (RIC). This evolved from the large multicentre CONDI-2-ERIC-PPCI outcome study which demonstrated no additional benefit when using RIC in the setting of ST-elevation myocardial infarction (STEMI). The workshop discussed how conditioning has led to a significant and fundamental understanding of the mechanisms preventing cell death following ischaemia and reperfusion, and the key target cyto-protective pathways recruited by protective interventions, such as RIC. However, the obvious need to translate this protection to the clinical setting has not materialised largely due to the disconnect between preclinical and clinical studies. Discussion points included how to adapt preclinical animal studies to mirror the patient presenting with an acute myocardial infarction, as well as how to refine patient selection in clinical studies to account for co-morbidities and ongoing therapy. These latter scenarios can modify cytoprotective signalling and need to be taken into account to allow for a more robust outcome when powered appropriately. The workshop also discussed the potential for RIC in other disease settings including ischaemic stroke, cardio-oncology and COVID-19. The workshop, therefore, put forward specific classifications which could help identify so-called responders vs. non-responders in both the preclinical and clinical settings.

The Landscape of Videofluoroscopy in the UK: A Web-Based Survey.

Videofluoroscopy (VFS) is considered one of the gold-standard assessments of swallowing. Whilst guidelines for the application and conduct of VFS exist, their translation into clinical practice remain challenging. To build a greater understanding on how VFS clinics operate in the UK. A web-based survey was shared with speech and language therapists (SLTs) working in VFS clinics via professional networks and social media from October 2018 to January 2019. 101 responses were received. Two thirds of clinics were SLT-led, with the majority of clinics being run by two SLTs (73.6%) and a radiographer (95.5%) also known as radiologic technologists, diagnostic radiographers and medical radiation technologists. Less than 50% of radiographers had received specialist training. Around half of the clinics used a standard assessment or analysis protocol and 88.1% a rating scale. Set recipes for a range of textures were used in 53.4% of VFS clinics. Barium and water soluble contrasts were used, but only 15.8% knew the concentration of contrast used. The most commonly reported VFS pulse and frame rate was 15 per second. There was evidence of a lack of SLT knowledge regarding technical operation of VFS. Screening times varied from 0.7-10 min (median 3 min, IQR 2.5-3.5). Around 50% of respondents reported quality issues affecting analysis. In a survey of UK SLTs, translation of VFS guidance into practice was found to be limited which may impact on the quality of assessment and analysis. Collaboration with radiology, strengthening of guidelines and greater uptake of specialist training is deemed essential.

Single-Shot Readout Performance of Two Heterojunction-Bipolar-Transistor Amplification Circuits at Millikelvin Temperatures.

High-fidelity single-shot readout of spin qubits requires distinguishing states much faster than the T1 time of the spin state. One approach to improving readout fidelity and bandwidth (BW) is cryogenic amplification, where the signal from the qubit is amplified before noise sources are introduced and room-temperature amplifiers can operate at lower gain and higher BW. We compare the performance of two cryogenic amplification circuits: a current-biased heterojunction bipolar transistor circuit (CB-HBT), and an AC-coupled HBT circuit (AC-HBT). Both circuits are mounted on the mixing-chamber stage of a dilution refrigerator and are connected to silicon metal oxide semiconductor (Si-MOS) quantum dot devices on a printed circuit board (PCB). The power dissipated by the CB-HBT ranges from 0.1 to 1 µW whereas the power of the AC-HBT ranges from 1 to 20 µW. Referred to the input, the noise spectral density is low for both circuits, in the 15 to 30 fA/[Formula: see text] range. The charge sensitivity for the CB-HBT and AC-HBT is 330 µe/[Formula: see text] and 400 µe/[Formula: see text], respectively. For the single-shot readout performed, less than 10 µs is required for both circuits to achieve bit error rates below 10-3, which is a putative threshold for quantum error correction.

A systematic review of cannabidiol dosing in clinical populations.

AIMS: Cannabidiol (CBD) is a cannabis-derived medicinal product with potential application in a wide-variety of contexts; however, its effective dose in different disease states remains unclear. This review aimed to investigate what doses have been applied in clinical populations, in order to understand the active range of CBD in a variety of medical contexts. METHODS: Publications involving administration of CBD alone were collected by searching PubMed, EMBASE and ClinicalTrials.gov. RESULTS: A total of 1038 articles were retrieved, of which 35 studies met inclusion criteria covering 13 medical contexts. Twenty-three studies reported a significant improvement in primary outcomes (e.g. psychotic symptoms, anxiety, seizures), with doses ranging between <1 and 50 mg/kg/d. Plasma concentrations were not provided in any publication. CBD was reported as well tolerated and epilepsy was the most frequently studied medical condition, with all 11 studies demonstrating positive effects of CBD on reducing seizure frequency or severity (average 15 mg/kg/d within randomised controlled trials). There was no signal of positive activity of CBD in small randomised controlled trials (range n = 6-62) assessing diabetes, Crohn's disease, ocular hypertension, fatty liver disease or chronic pain. However, low doses (average 2.4 mg/kg/d) were used in these studies. CONCLUSION: This review highlights that CBD has a potential wide range of activity in several pathologies. Pharmacokinetic studies as well as conclusive phase III trials to elucidate effective plasma concentrations within medical contexts are severely lacking and highly encouraged.

The dilation aided single-line-of-sight x-ray camera for the National Ignition Facility: Characterization and fielding.

Crystal x-ray imaging is frequently used in inertial confinement fusion and laser-plasma interaction applications as it has advantages compared to pinhole imaging, such as higher signal throughput, better achievable spatial resolution, and chromatic selection. However, currently used x-ray detectors are only able to obtain a single time resolved image per crystal. The dilation aided single-line-of-sight x-ray camera described here was designed for the National Ignition Facility (NIF) and combines two recent diagnostic developments, the pulse dilation principle used in the dilation x-ray imager and a ns-scale multi-frame camera that uses a hold and readout circuit for each pixel. This enables multiple images to be taken from a single-line-of-sight with high spatial and temporal resolution. At the moment, the instrument can record two single-line-of-sight images with spatial and temporal resolution of 35 µm and down to 35 ps, respectively, with a planned upgrade doubling the number of images to four. Here we present the dilation aided single-line-of-sight camera for the NIF, including the x-ray characterization measurements obtained at the COMET laser, as well as the results from the initial timing shot on the NIF.

Estimating the risk of exposure of British livestock to foot-and-mouth disease associated with the importation of ship and aircraft waste.

A quantitative risk assessment model was developed to estimate the frequency with which meat waste from ships or aircraft might expose British livestock to infection with foot-and-mouth disease (fmd), and investigate the factors that might contribute to the risk. In the model, the total weight of waste introduced that was contaminated with fmd was estimated to be 26 kg per year, with 90 per cent certainty that it would be between 10 and 53 kg. As a result, it was estimated that there would be a mean of 1429 years between outbreaks of fmd due to ship and aircraft waste, with a 90 per cent certainty that the interval would be between 500 and 10,000 years. These estimates were affected by three principal uncertainties: first, the prevalence of fmd; secondly, the probability of the waste being removed and fed to pigs; and thirdly, the probability that overboard dumping of contaminated waste might expose livestock to fmd. The effect of these uncertainties on the model was investigated in a sensitivity analysis.

A quantitative risk assessment for the importation of brucellosis-infected breeding cattle into Great Britain from selected European countries.

Great Britain (GB) has been "Officially Brucellosis Free" (OBF) since 1991; because this disease has both public-health and international-trade implications, it is in the country's interest to maintain this freedom. A quantitative risk-assessment model was developed to determine the annual risk of importing brucellosis-infected breeding cattle into GB from Northern Ireland and the Republic of Ireland. (These countries exported the largest number of cattle into GB and were not brucellosis free during the development of the assessment in 2000.) We predicted that we can expect to import brucellosis from Northern Ireland every 2.63 years (1.89, 4.17) and from the Republic of Ireland, every 3.23 years (2.13, 5.88). The estimates of risk are sensitive to the assumed proportion of animals missed during routine surveillance that originate from OBF herds and the uncertainty associated with the surveillance test sensitivities. As a result of the assessment, the Department for Environment, Food and Rural Affairs (Defra) introduced post-calving testing for all cattle imported into British herds.

A simulation model of brucellosis spread in British cattle under several testing regimes.

Brucellosis is a widespread, economically devastating and highly infectious zoonosis. In cattle, infection predominantly is caused by Brucella abortus, and is usually detected in pregnant females through abortions. Great Britain (GB) has been declared free from brucellosis (officially brucellosis free (OBF)) since 1993 and as such is required by European Union (EU) regulations to test > or =20% of both beef and dairy cattle >24 months old routinely. Currently, however, GB serologically tests more cattle than required and the issue of reducing the level of testing has come under consideration. We developed a simulation model to determine the rate of spread of brucellosis under a variety of testing regimes. For dairy herds, we found that reducing the level of testing would have a major effect on the rate of spread of infection, should it be imported. For beef herds, reducing the level of testing would have much less effect. We also found that abortion notification is a very-important additional means of surveillance. As a result of our predictions, policy-makers decided not to reduce the level of testing and actively to promote abortion notification.

Quantitative estimates of the risk of new outbreaks of foot-and-mouth disease as a result of burning pyres.

The risk of dispersing foot-and-mouth disease virus into the atmosphere, and spreading it to susceptible holdings as a result of burning large numbers of carcases together on open pyres, has been estimated for six selected pyres burned during the 2001 outbreak in the UK. The probability of an animal or holding becoming infected was dependent on the estimated level of exposure to the virus predicted from the concentrations of virus calculated by the Met Office, Bracknell. In general, the probability of infection per animal and per holding decreased as their distance from the pyre increased. In the case of two of the pyres, a holding under the pyre plumes became infected on a date consistent with when the pyre was lit. However, by calculating their estimated probability of infection from the pyres it was concluded that it was unlikely that in either case the pyre was the source of infection.

Iron supplementation and oxidative damage to DNA in healthy individuals with high plasma ascorbate.

Previously, we have investigated the potential for a pro-oxidant interaction of iron and ascorbate in vivo in iron and ascorbate cosupplementation or ascorbate supplementation studies. In this study, for the first time, the effects of iron supplementation on oxidative damage to DNA in healthy individuals with plasma ascorbate levels at the upper end of the normal range were examined. Forty female and male volunteers (mean plasma ascorbate approximately equal to 70 micromol/L) were supplemented with a daily dose of syrup (ferrous glycine sulphate equivalent to 12.5 mg iron) for 6 weeks. Serum ferritin, transferrin bound iron, % saturation of transferrin and plasma ascorbate were assessed and the mean dietary intakes of all subjects were estimated through food frequency questionnaires. Oxidative damage to DNA bases from white blood cells was measured by gas chromatography/mass spectrometry with selected-ion monitoring (GC/MS-SIM), using isotope-labelled standards for quantification. Iron supplementation did not affect any of the iron status parameters. There were also no detrimental effects, over the period under investigation, in terms of oxidative damage to DNA. However, the effects of larger doses or of longer supplementation periods should also be investigated.

The steady-state levels of oxidative DNA damage and of lipid peroxidation (F2-isoprostanes) are not correlated in healthy human subjects.

Oxidative damage to DNA in human tissues can be determined by measuring multiple products of oxidative damage to the purine and pyrimidine bases using gas chromatography-mass spectrometry (GC-MS). Oxidative damage to lipids (lipid peroxidation) can be quantitated by the mass spectrometry-based determination of F2-isoprostanes, specific end-products of the peroxidation of arachidonic acid residues in lipids. For both DNA base damage products and 8-epi prostaglandin F2alpha (PGF2alpha), there is a wide variation in levels between different healthy human subjects. We measured multiple products of oxidative damage to DNA bases in white cells, and 8-epi PGF2alpha in plasma, from blood samples obtained from healthy human subjects in the UK and in Portugal. No correlation of 8-epi PGF2alpha levels with levels of any modified DNA base (including 8-hydroxyguanine) was observed. We conclude that no single parameter can be measured as an index of "oxidative stress" or "oxidative damage" in vivo.

Effect of dietary quercetin on oxidative DNA damage in healthy human subjects.

The effect of dietary intake of flavonols (predominantly quercetin) on oxidative DNA damage was studied in thirty-six healthy human subjects (sixteen men, twenty women). The study was a randomised crossover study, comprising two 14 d treatments of either a low-flavonol (LF) or high-flavonol (HF) diet with a 14 d wash-out period between treatments. Subjects were asked to avoid foods containing flavonols, flavones and flavanols during the LF dietary treatment period and to consume one 150 g onion (Allium cepa) cake (containing 89.7 mg quercetin) and one 300 ml cup of black tea (containing 1.4 mg quercetin) daily during the HF dietary treatment. A 7 d food diary was kept during each dietary period and blood samples were taken after each dietary treatment. Products of oxidative damage to DNA bases were measured in DNA from leucocytes. The study had more than 95% power to detect a change of 20% in DNA damage products Plasma vitamin C and plasma quercetin concentrations were also measured. No significant differences in intake of macronutrients or assessed micronutrients, measured DNA base damage products, or plasma vitamin C were found between the HF and LF dietary treatments. The plasma quercetin concentration was significantly higher after the HF dietary treatment period (228.5 (SEM 34.7) nmol/l) than after the LF dietary treatment period (less than the limit of detection, i.e. <66.2 nmol/l). These findings do not support the hypothesis that dietary quercetin intake substantially affects oxidative DNA damage in leucocytes.

Analytic solutions to compartmental models of the HIV/AIDS epidemic.

For many years compartmental models have provided useful insights into the spread of epidemics. Such models are usually fairly easy to set up, but even the simpler models have the disadvantage that they are intractable to analytic solution. In this paper we examine models of the HIV/AIDS epidemic, and show that the equations may be linearized in a piecewise manner over time, thus allowing analytic solutions to be obtained. Indications of the usefulness of this approach are provided. In particular, an analytic solution gives insight into the mechanism of the epidemic, together with a clearer picture of the sensitivity of results to changes in parameter values. Further, the processes of parameter estimation and the methodology of back-calculation also benefit from the provision of functional forms for the state variables.

Determination of oxidative DNA base damage by gas chromatography-mass spectrometry. Effect of derivatization conditions on artifactual formation of certain base oxidation products.

GC-MS is a widely used tool to measure oxidative DNA damage because of its ability to identify a wide range of base modification products. However, it has been suggested that the derivatization procedures required to form volatile products prior to GC-MS analysis can sometimes produce artifactual formation of certain base oxidation products, although these studies did not replicate previously-used reaction conditions, e.g. they failed to remove air from the derivatization vials. A systematic examination of this problem revealed that levels of 8-hydroxyguanine, 8-hydroxyadenine, 5-hydroxycytosine and 5-(hydroxymethyluracil) in commercial calf thymus DNA determined by GC-MS are elevated by increasing the temperature at which derivatization is performed in our laboratory. In particular, 8-hydroxyguanine levels after silylation at 140 degrees C were raised 8-fold compared to derivatization at 23 degrees C. Experiments on the derivatization of each undamaged base revealed that the artifactual oxidation of guanine, adenine, cytosine and thymine respectively was responsible. Formation of the above products was potentiated by not purging with nitrogen prior to derivatization. Increasing the temperature to 140 degrees C or allowing air to be present during derivatization did not significantly increase levels of the other oxidized bases measured. This work suggests that artifactual oxidation during derivatization is restricted to certain products (8-hydroxyguanine, 8-hydroxyadenine, 5-hydroxycytosine and 5-[hydroxymethyluracil]) and can be decreased by reducing the temperature of the derivatization reaction to 23 degrees C and excluding as much air possible. Despite some recent reports, we were easily able to detect formamidopyrimidines in acid-hydrolyzed DNA. Artifacts of derivatization are less marked than has been claimed in some papers and may vary between laboratories, depending on the experimental procedures used, in particular the efficiency of exclusion of O2 during the derivatization process.

Measurement of oxidative DNA damage by gas chromatography-mass spectrometry: ethanethiol prevents artifactual generation of oxidized DNA bases.

Analysis of oxidative damage to DNA bases by GC-MS enables identification of a range of base oxidation products, but requires a derivatization procedure. However, derivatization at high temperature in the presence of air can cause 'artifactual' oxidation of some undamaged bases, leading to an overestimation of their oxidation products, including 8-hydroxyguanine. Therefore derivatization conditions that could minimize this problem were investigated. Decreasing derivatization temperature to 23 degrees C lowered levels of 8-hydroxyguanine, 8-hydroxyadenine, 5-hydroxycytosine and 5-(hydroxymethyl)uracil measured by GC-MS in hydrolysed calf thymus DNA. Addition of the reducing agent ethanethiol (5%, v/v) to DNA samples during trimethylsilylation at 90 degrees C also decreased levels of these four oxidized DNA bases as well as 5-hydroxyuracil. Removal of guanine from hydrolysed DNA samples by treatment with guanase, prior to derivatization, resulted in 8-hydroxyguanine levels (54-59 pmol/mg of DNA) that were significantly lower than samples not pretreated with guanase, independent of the derivatization conditions used. Only hydrolysed DNA samples that were derivatized at 23 degrees C in the presence of ethanethiol produced 8-hydroxyguanine levels (56+/-8 pmol/mg of DNA) that were as low as those of guanase-pretreated samples. Levels of other oxidized bases were similar to samples derivatized at 23 degrees C without ethanethiol, except for 5-hydroxycytosine and 5-hydroxyuracil, which were further decreased by ethanethiol. Levels of 8-hydroxyguanine, 8-hydroxyadenine and 5-hydroxycytosine measured in hydrolysed calf thymus DNA by the improved procedures described here were comparable with those reported previously by HPLC with electrochemical detection and by GC-MS with prepurification to remove undamaged base. We conclude that artifactual oxidation of DNA bases during derivatization can be prevented by decreasing the temperature to 23 degrees C, removing air from the derivatization reaction and adding ethanethiol.

Spontaneous preference for ethanol in naive rats is influenced by cholecystokinin A receptor antagonism.

Naive adult male Wistar rats free to choose between water or 10% ethanol (v/v) spontaneously became water-preferring (WP) rats, as they drank mainly water (approximately 35 ml per day), or alcohol-drinking (ED) rats, as they also drank a significant amount of ethanol (approximately 14 ml per day). The selective CCKA receptor antagonist L-364,718 at doses selective for the CCKA receptor (5 micrograms/kg, IP) halved the consumption of alcohol of the ED rats without modifying their total liquid in-take. In contrast, the CCKB antagonists L-365,260 or GV150013 were without effect when used at doses selective for the CCKB receptor. These data indicate that the CCK system could be involved in the modulation of alcohol intake. In particular, they suggest that CCKA receptors could play a role in the ethanol preference.

Antioxidant action of ergothioneine: assessment of its ability to scavenge peroxynitrite.

The superoxide radical (O.2-) and nitric oxide (NO.) combine very rapidly to form peroxynitrite (ONOO-), a reactive tissue damaging nitrogen species thought to be involved in the pathology of several chronic diseases. The natural product ergothioneine protects against the nitration of tyrosine and the inactivation of alpha 1-antiproteinase by ONOO-. Ergothioneine merits further investigation as a biological and therapeutic antioxidant agent.

Simultaneous, selective detection of catecholaminergic and indolaminergic signals using cyclic voltammetry with treated micro-sensor.

Selective and simultaneous voltammetric analysis of catechols and indoles in vivo and in vitro has until now been feasible only by means of 'slow' scanning methods (scan speed in tens of seconds) such as differential pulse (DPV) and differential normal pulse voltammetry in conjunction with electrically and/or chemically treated carbon-fiber micro-electrodes (mCFE). Faster electrochemical techniques, such as chronoamperometry and cyclic voltammetry (CV), allow more rapid (seconds or fractions of a second) and frequent measurements of these chemicals. However, these methods show poor sensitivity and selectivity in the presence of different electroactive compounds with similar oxidation potentials. In order to analyze whether the lack of sensitivity and selectivity of the fast voltammetric methods results from the rapidity of the measurement or from the use of untreated sensors, the methods of CV (scan speed: 1000 mV/s) and DPV (scan speed: 10 mV/s) have been applied with either untreated or electrically treated mCFE to analyze the in vitro oxidation potential and current values of DA and 5-HT. When associated with untreated mCFE, neither method was able to separate and selectively detect the two compounds dissolved together in an inert vehicle; the voltammogram recorded resulted in a single broad oxidation signal. In contrast, when these techniques were performed with electrically treated mCFE, oxidation signals for DA (peak A) and 5-HT (peak B) were monitored simultaneously at approximately + 65 mV and + 240 mV, with DPV respectively, and at + 120 mV and + 300 mV with CV, respectively. Additionally, CV with treated mCFE on anesthetized rats, simultaneously monitored two striatal signals at approximately + 100 mV and + 300 mV. The oxidation values (Em) and current levels (nA) of these peaks remained stable during control recordings. The current levels were selectively increased by peripheral injection of fluphenazine (DA antagonist) or of 5-hydroxytryptophan (precursor of serotonin). The chemical nature of these two peaks may therefore be considered catecholaminergic and indolaminergic, respectively. Hence, this report provides the first evidence for the feasibility of concomitant in vitro analysis of DA and 5-HT using a rapid scanning method such as CV. In addition, the values of current level (nA) obtained with CV-mCFE for DA and 5-HT are comparable to those monitored with DPV-mCFE, supporting the view that treatment of the sensor is a key point for increasing the selectivity and the sensitivity of these voltammetric techniques. The feasibility of using CV with electrically treated mCFE for fast in vivo analysis of catechol and indole activities is also demonstrated.

Carbon fibre micro-electrodes for concomitant in vivo electrophysiological and voltammetric measurements: no reciprocal influences.

Differential pulse voltammetry and more recently cyclic voltammetry have been successfully used to monitor basal levels of endogenous chemicals by means of treated carbon fibre microbiosensors inserted in specific brain regions. In this study, feasibility of concomitant in vivo recordings of stable electrophysiological signals and basal ascorbate, catecholaminergic and indolaminergic voltammetric peaks at the same cerebral site by means of a single electrically treated carbon fibre micro electrode (microbiosensor) is presented. The results indicate that these two independent techniques can be combined in vivo at a single electrode, and that voltammetric measurements of unstimulated levels of extracellular compounds do not alter concomitant basal cell firing for a period long enough (more than 6 h) to allow pharmacological manipulations.

Identification of the diastereomers of pentobarbital N-glucosides excreted in human urine.

A study was undertaken to determine if humans excreted pentobarbital N-glucosides as urinary metabolites following oral administration of pentobarbital. (1'RS,5RS)-1-(beta-D-Glucopyranosyl)pentobarbital ((1'RS,5RS)-PTBG) was isolated from the urine of one subject. The two diastereomers, (1'RS,5R)-PTBG and (1'RS,5S)-PTBG were separated and found to be identical to synthetic standards when compared using HPLC retention times coupled with UV (with and without post-column ionization) and mass spectrometry (HPLC/MS). A HPLC method was developed for detecting and quantifying (1'RS,5R)-PTBG, (1'RS,5S)-PTBG and pentobarbital in urine. Following a single oral dose of sodium pentobarbital to male subjects (n = 6), 1.6-6.2% of the pentobarbital dose was excreted as (1'RS,5S)-PTBG over 60 hours. (1'RS,5R)-PTBG was also detected in one subject and accounted for 0.3% of the pentobarbital dose. Using a modified HPLC system, the four pentobarbital N-glucosides were resolved and analysis of a partially purified pentobarbital N-glucoside extract from one subject indicated that only (1'R,5R)-PTBG and (1'S,5S)-PTBG could be detected as urinary excretion products. These results indicate that the side chain chirality of pentobarbital may influence the observed enantioselectivity for the formation and/or urinary excretion of the pentobarbital N-glucosides.