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Summary: Maximizing the personal, public, research, and clinical value of genomic information will require the reliable exchange of genetic variation data. We report here the Variation Representation Specification (VRS, pronounced “verse”), an extensible framework for the computable representation of variation that complements contemporary human-readable and flat file standards for genomic variation representation. VRS provides semantically precise representations of variation and leverages this design to enable federated identification of biomolecular variation with globally consistent and unique computed identifiers. The VRS framework includes a terminology and information model, machine-readable schema, data sharing conventions, and a reference implementation, each of which is intended to be broadly useful and freely available for community use. VRS was developed by a partnership among national information resource providers, public initiatives, and diagnostic testing laboratories under the auspices of the Global Alliance for Genomics and Health (GA4GH).

Summary: The Global Alliance for Genomics and Health (GA4GH) aims to accelerate biomedical advances by enabling the responsible sharing of clinical and genomic data through both harmonized data aggregation and federated approaches. The decreasing cost of genomic sequencing (along with other genome-wide molecular assays) and increasing evidence of its clinical utility will soon drive the generation of sequence data from tens of millions of humans, with increasing levels of diversity. In this perspective, we present the GA4GH strategies for addressing the major challenges of this data revolution. We describe the GA4GH organization, which is fueled by the development efforts of eight Work Streams and informed by the needs of 24 Driver Projects and other key stakeholders. We present the GA4GH suite of secure, interoperable technical standards and policy frameworks and review the current status of standards, their relevance to key domains of research and clinical care, and future plans of GA4GH. Broad international participation in building, adopting, and deploying GA4GH standards and frameworks will catalyze an unprecedented effort in data sharing that will be critical to advancing genomic medicine and ensuring that all populations can access its benefits.

To analyze the cause of the destruction of thin, carbon-backed lithium fluoride targets during a measurement of the fusion of 7Li and 17O, we estimate theoretically the lifetimes of carbon and LiF films due to sputtering, thermal evaporation, and lattice damage and compare them with the lifetime observed in the experiment. Sputtering yields and thermal evaporation rates in carbon and LiF films are too low to play significant roles in the destruction of the targets. We estimate the lifetime of the target due to lattice damage of the carbon backing and the LiF film using a previously reported model. In the experiment, elastically scattered target and beam ions were detected by surface silicon barrier (SSB) detectors so that the product of the beam flux and the target density could be monitored during the experiment. The areas of the targets exposed to different beam intensities and fluences were degraded and then perforated, forming holes with a diameter around the beam spot size. Overall, the target thickness tends to decrease linearly as a function of the beam fluence. However, the thickness also exhibits an increasing interval after SSB counts per beam ion decreases linearly, extending the target lifetime. The lifetime of thin LiF film as determined by lattice damage is calculated for the first time using a lattice damage model, and the calculated lifetime agrees well with the observed target lifetime during the experiment. In experiments using a thin LiF target to induce nuclear reactions, this study suggests methods to predict the lifetime of the LiF film and arrange the experimental plan for maximum efficiency.

Patients with Alzheimer's disease (AD) often have fragmentation of sleep/wake cycles and disrupted 24-h (circadian) activity. Despite this, little work has investigated the potential underlying day/night disruptions in cognition and neuronal physiology in the hippocampus. The molecular clock, an intrinsic transcription-translation feedback loop that regulates circadian behavior, may also regulate hippocampal neurophysiological activity. We hypothesized that disrupted diurnal variation in clock gene expression in the hippocampus corresponds with loss of normal day/night differences in membrane excitability, synaptic physiology, and cognition. We previously reported disrupted circadian locomotor rhythms and neurophysiological output of the suprachiasmatic nucleus (the primary circadian clock) in Tg-SwDI mice with human amyloid-beta precursor protein mutations. Here, we report that Tg-SwDI mice failed to show day/night differences in a spatial working memory task, unlike wild-type controls that exhibited enhanced spatial working memory at night. Moreover, Tg-SwDI mice had lower levels of Per2, one of the core components of the molecular clock, at both mRNA and protein levels when compared to age-matched controls. Interestingly, we discovered neurophysiological impairments in area CA1 of the Tg-SwDI hippocampus. In controls, spontaneous inhibitory post-synaptic currents (sIPSCs) in pyramidal cells showed greater amplitude and lower inter-event interval during the day than the night. However, the normal day/night differences in sIPSCs were absent (amplitude) or reversed (inter-event interval) in pyramidal cells from Tg-SwDI mice. In control mice, current injection into CA1 pyramidal cells produced more firing during the night than during the day, but no day/night difference in excitability was observed in Tg-SwDI mice. The normal day/night difference in excitability in controls was blocked by GABA receptor inhibition. Together, these results demonstrate that the normal diurnal regulation of inhibitory transmission in the hippocampus is diminished in a mouse model of AD, leading to decreased daytime inhibition onto hippocampal CA1 pyramidal cells. Uncovering disrupted day/night differences in circadian gene regulation, hippocampal physiology, and memory in AD mouse models may provide insight into possible chronotherapeutic strategies to ameliorate Alzheimer's disease symptoms or delay pathological onset.

Summary: Feeding rodents a high-fat diet (HFD) disrupts normal behavioral rhythms, particularly meal timing. Within the brain, mistimed feeding shifts molecular rhythms in the hippocampus and impairs memory. We hypothesize that altered meal timing induced by an HFD leads to cognitive impairment and that restricting HFD access to the “active period” (i.e., night) rescues the normal hippocampal function. In male mice, ad-lib access to an HFD for 20 weeks increased body weight and fat mass, increased daytime meal consumption, reduced hippocampal long-term potentiation (LTP), and eliminated day/night differences in spatial working memory. Importantly, two weeks of time-restricted feeding (TRF) at the end of the chronic HFD protocol rescued spatial working memory and restored LTP magnitude, even though there was no change in body composition and total daily caloric intake. These findings suggest that short-term TRF is an effective mechanism for rescuing HFD-induced impaired cognition and hippocampal function.

With the move away from safety testing assessment based on data generated in experimental animals the concept of Next Generation Risk Assessment (NGRA) has arisen which instead uses data from in silico and in vitro models. A key uncertainty in risk assessment is the actual dose of test chemical at the target site, and therefore surrogate dose metrics, such as nominal concentration in test media are used to describe in vitro effect (or no-effect) doses. The reliability and accuracy of the risk assessment therefore depends largely on our ability to understand and characterise the relationship between the dose metrics used and the actual biologically effective dose at the target site. The objective of this publication is to use 40 case study chemicals to illustrate how in vitro dose considerations can be applied to characterise the "true dose" and build confidence in the understanding of the biologically effective dose in in vitro test systems for the determination e.g. points of departure (PoDs) for NGRA. We propose a workflow that can be applied to assess whether the nominal test concentration can be considered a conservative dose metric for use in NGRA. The workflow examines the implications of volatility, stability, hydrophobicity, binding to plastic and serum, solubility, and the potential use of in silico models for some of these parameters. For the majority of the case study chemicals we found that the use of nominal concentrations in risk assessment would result in conservative decision making. However, for serval chemicals a potential for underestimation of the risk in humans in vivo based on in vitro nominal effect concentrations was identified, and approaches for refinement by characterisation of the actual effect concentration are proposed., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Cardiopulmonary exercise testing (CPET) identifies high-risk patients before major surgery. In addition to using oxygen uptake and ventilatory efficiency to assess functional capacity, CPET can be used to identify underlying myocardial dysfunction through the assessment of the oxygen uptake to heart rate response (oxygen pulse response). We examined the relationship of oxygen pulse response, in combination with other CPET variables and known cardiac risk factors, with mortality after colorectal cancer surgery., Methods: This work focused on a retrospective cohort study of patients who had CPET and underwent colorectal cancer surgery. The primary outcome was a composite of in-hospital and 30-day mortality. Ventilatory inefficiency (Ve/Vco 2 >34) and exercise-induced myocardial dysfunction (abnormal oxygen pulse response) were investigated for an association with mortality using bivariable analysis and multivariable Cox regression., Results: A total of 1214 patients who underwent colorectal cancer surgery were included, and the primary outcome occurred in 26 patients (2.1%). Multivariable Cox regression showed abnormal oxygen pulse response was independently associated with the primary outcome (odds ratio [OR]=2.75; 95% confidence interval [CI], 1.17-6.47). Bivariable analysis showed that Ve/Vco 2 >34 was associated with the primary outcome (OR=3.43; 95% CI, 1.47-8.01). Combining Ve/Vco 2 >34 and abnormal oxygen pulse response conferred an increased risk for the primary outcome (OR=4.47; 95% CI, 1.62-12.34), compared with Ve/Vco 2 >34 and normal oxygen pulse response., Conclusion: Ventilatory inefficiency and an abnormal oxygen pulse response were independently associated with short- (30-day) and long-term (2-yr) mortality. Oxygen pulse response may provide additional information when considering perioperative risk stratification., (Copyright © 2020 British Journal of Anaesthesia. Published by Elsevier Ltd. All rights reserved.)

Psoriasis increases the risk of cardiovascular (CV) disease. Secukinumab, a fully human monoclonal antibody against IL-17A, shows significant efficacy in psoriasis, but effects on CV markers are unknown. CARIMA (Evaluation of Cardiovascular Risk Markers in Psoriasis Patients Treated with Secukinumab) was a 52-week, randomized, double-blind, placebo-controlled, exploratory trial in patients with moderate to severe plaque psoriasis without clinical CV disease. Patients were randomly assigned to receive 300 mg or 150 mg secukinumab until week 52 or to receive placebo until week 12 and then 300 mg or 150 mg secukinumab until week 52. The primary outcome was endothelial function measured by flow-mediated dilation (FMD). Baseline FMD was significantly lower in psoriasis patients than healthy volunteers (4.4 ± 3.9% vs. 6.1 ± 3.3%, P = 0.01). At week 12, baseline-adjusted mean FMD was numerically higher in patients receiving secukinumab versus those receiving placebo, but this difference (300-mg group, +1.2%; 150-mg group, +0.76%; P = 0.223 and P = 0.403 by analysis of covariance) did not reach significance. At week 52, FMD increased across groups. FMD was significantly higher than baseline in patients receiving the label dose of 300 mg secukinumab for 52 weeks (+2.1%, 95% confidence interval = 0.8-3.3; P = 0.0022). Other relevant CV markers were unchanged. CARIMA indicates that secukinumab might have a beneficial effect on CV risk by improving the endothelial function of patients with plaque psoriasis., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

Competing Interests: The authors declare that they have no conflicts of interest.

Objective: To compare the clinical effects of alfaxalone, ketamine and propofol in dogs following premedication with medetomidine and methadone., Study Design: Prospective, 'blinded' and randomized clinical study., Animals: A total of 75 male dogs presented for neutering at a charity clinic., Methods: Dogs were allocated to be administered alfaxalone, ketamine or propofol following premedication with medetomidine (20 μg kg -1 ) and methadone (0.2 mg kg -1 ). Dogs were temperament scored prior to premedication. Quality of sedation, induction of anaesthesia, recovery and recovery environment were scored by simple descriptive scales. Physiological variables during anaesthesia were recorded. Continuous numerical data were analysed using analysis of variance with repeated measures as necessary. Nonparametric data were analysed using Kruskal-Wallis tests and multiple comparisons using Dunn's test. Statistical significance was set at p < 0.05., Results: The mean (± standard deviation) dose of alfaxalone was 0.6 ± 0.2 mg kg -1 , that for ketamine was 1.5 ± 0.7 mg kg -1 and that for propofol was 0.8 ± 0.3 mg kg -1 . Alfaxalone inductions were significantly smoother compared to ketamine but not to propofol. Only one of 75 of the inductions was deemed poor. There were no differences in cardiopulmonary variables between groups except immediately after induction of anaesthesia. There were no differences in quality of recovery between groups., Conclusions and Clinical Relevance: All three induction agents provided reliable, predictable anaesthesia conditions that were clinically indistinguishable and ideal for teaching anaesthesia skills. The medetomidine and methadone premedication resulted in profound, heavy sedation and the quality of induction of anaesthesia was better with alfaxalone compared to ketamine. No significant difference in induction quality was detected between alfaxalone and proprofol or propofol and ketamine, and these findings are likely to be of limited clinical significance when choosing an induction agent., (Copyright © 2017 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

I.V. fluid therapy plays a fundamental role in the management of hospitalized patients. While the correct use of i.v. fluids can be lifesaving, recent literature demonstrates that fluid therapy is not without risks. Indeed, the use of certain types and volumes of fluid can increase the risk of harm, and even death, in some patient groups. Data from a recent audit show us that the inappropriate use of fluids may occur in up to 20% of patients receiving fluid therapy. The delegates of the 12th Acute Dialysis Quality Initiative (ADQI) Conference sought to obtain consensus on the use of i.v. fluids with the aim of producing guidance for their use. In this article, we review a recently proposed model for fluid therapy in severe sepsis and propose a framework by which it could be adopted for use in most situations where fluid management is required. Considering the dose-effect relationship and side-effects of fluids, fluid therapy should be regarded similar to other drug therapy with specific indications and tailored recommendations for the type and dose of fluid. By emphasizing the necessity to individualize fluid therapy, we hope to reduce the risk to our patients and improve their outcome., (© The Author 2014. Published by Oxford University Press on behalf of the British Journal of Anaesthesia. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Study Objective: To compare stroke volume (SV) and preload responsiveness measurements from different technologies with the esophageal Doppler monitor (EDM)., Design: Prospective measurement study., Setting: Operating room., Patients: 20 ASA physical status 3 patients undergoing vascular, major urological, and bariatric surgery., Interventions: Subjects received fluids using a standard Doppler protocol of 250 mL of colloid administered until SV no longer increased by >10%, and again when the measured SV decreased by 10%., Measurements: Simultaneous readings of SV, stroke volume variation (SVV) and pulse pressure variation (PPV) from the LiDCOrapid, and SVV from the FloTrac/Vigileo were compared with EDM measurements. The pleth variability index (PVI) also was recorded., Main Results: No correlation was seen in percentage SV change as measured by either the LiDCOrapid (r=0.05, P=0.616) or FloTrac (r=0.09, P= 0.363) systems compared with the EDM. Correlation was present between the LiDCOrapid and FloTrac (r=0.515, P<0.0001). Percentage error compared with the EDM was 81% for the FloTrac and 90% for the LiDCOrapid. SVV as measured by LiDCOrapid differed for fluid responders and nonresponders (10% vs 7%; P=0.021). Receiver operator curve analysis to predict a 10% increase in SV from the measured variables showed an area under the curve of 0.57 (95% CI 0.43-0.72) for SVV(FloTrac), 0.64 (95% CI 0.52-0.78) for SVV(LiDCO), 0.61 (95% CI 0.46 -0.76) for PPV, and 0.59 (95% CI 0.46 -0.71) for PVI., Conclusions: Stroke volume as measured by the FloTrac and LiDCOrapid systems does not correlate with the esphageal Doppler, has poor concordance, and a clinically unacceptable percentage error. The predictive value of the fluid responsiveness parameters is low, with only SVV measured by the LiDCOrapid having clinical utility., (© 2013 Elsevier Inc. All rights reserved.)

Background: Studies of preoperative cardiopulmonary exercise testing (CPET) have shown that a reduced oxygen uptake at anaerobic threshold (AT) and elevated ventilatory equivalent for carbon dioxide (VE/VCO(2)) were associated with reduced short- and medium-term survival after major surgery. The aim of this study was to determine the relative values of these, and also clinical risk factors, in identifying patients at risk of death after major intra-abdominal, non-vascular surgery., Methods: Patients aged >55 yr, undergoing elective colorectal resection, radical nephrectomy, or cystectomy between June 2004 and May 2009 had CPET during their routine pre-assessment clinic visit. We performed a retrospective analysis of known clinical risk factors and data from CPET to assess their relationship to all-cause mortality after surgery., Results: Eight hundred and forty-seven patients underwent surgery, of whom 18 (2.1%) died. A clinical history of ischaemic heart disease (RR 3.1, 95% CI 1.3-7.7), a VE/VCO(2) >34 (RR 4.6, 95% CI 1.4-14.8), and an AT < or =10.9 ml kg(-1) min(-1) (RR 6.8, 95% CI 1.6-29.5) were all significant predictors of all-cause hospital and 90 day mortality. The effect of reduced AT was most pronounced in patients with no history of cardiac risk factors (RR 10.0, 95% CI 1.7-61.0)., Conclusions: The routine measurement of AT and VE/VCO(2) using CPET for patients undergoing high-risk surgery can accurately identify the majority of high-risk patients, while the use of clinical risk factors alone will only identify a relatively small proportion of at-risk patients.

Ractopamine HCl and zilpaterol HCl, beta-agonists recently approved for use in feedlot cattle to improve performance traits and carcass leanness, were examined for their effects on fecal shedding of Escherichia coli O157:H7 in feedlot cattle. Fecal samples (n = 2,454) were obtained from four experiments (one ractopamine HCl, three zilpaterol HCl) over the course of a 3-year period, either by rectal palpation (ractopamine HCl experiment) or from pen-floor fecal pats. Samples were cultured quantitatively and qualitatively for E. coli O157:H7. No significant treatment differences were detected for fecal prevalence of E. coli O157:H7 in the ractopamine HCl experiment. Zilpaterol HCl feeding had no effect (P > 0.20) on fecal shedding in the first or second experiments, with overall E. coli O157:H7 prevalence relatively low (<7%). In the third zilpaterol HCl experiment, the percentage of fecal samples that were E. coli O157:H7 positive following qualitative culture was higher (P < 0.05) in the zilpaterol HCl treatment (10.3%) than for the control (6.1%). The current research showed minimal effects of beta-agonists on fecal shedding of E. coli O157:H7 and indicated that these compounds (fed immediately prior to slaughter) are not a cause for concern from a food safety standpoint.

Background: Body mass loss is inevitable with chronic hypoxic exposure. However, the exact body-composition changes, their causes, and possible treatments remain unknown., Objective: The objective was to investigate body composition during a high-altitude expedition by using non-empirically derived methods, experimentally manipulating energy intake, and investigating the influence of initial body composition., Design: Forty-one participants completed a 21-d expedition in the Himalayas. Energy intake was manipulated with a double-blind, placebo-controlled, randomized trial of carbohydrate energy supplementation. Body composition was assessed before and after the expedition by using a 4-component model including fat mass, total body water, bone mineral mass, and residual mass (principally protein and glycogen). Data were analyzed by repeated-measures analysis of variance., Results: Participants allocated to receive carbohydrate were given an additional 15,058 +/- 6211 kcal over the 21-d expedition (>6 kcal x kg(-1) x d(-1)). Nevertheless, the functionally important residual mass decreased in both groups by 6% (main effect of time: P = 0.021), with no effect of allocation (interaction effect: P = 0.116). Similar decreases were observed for fat mass (11%) and total body water (3%), which were also unabated by allocation. Furthermore, high initial fat mass (by median split) did not preserve residual mass (high-fat compared with low-fat participants: residual loss = 5% compared with 8%; P = 0.990)., Conclusions: High-altitude exposure decreased body mass, including the functionally important residual component. These losses were not abated by increasing energy intake or an initially high fat mass. Factors other than negative energy balance must contribute to body-composition changes with chronic hypoxia. This trial was registered at clinicaltrials.gov as NCT00731510.

Introduction: minimally invasive placement of intracardiac (IC) ECG leads in monkeys has greatly improved signal quality and the ability to interpret these ECGs. However, information on characteristics of the ECGs recorded using the IC lead is not available in the literature. There are concerns about the potential impact of IC lead placement on the ECG waveform and cardiac function as a result of potential irritation or trauma resulting from the placement and/or long term residence of the IC lead. The purposes of this study were to characterize IC ECG morphology, to obtain information on the recovery processes after IC ECG lead implantation, and to evaluate the IC ECG model application to safety pharmacology studies., Methods: the telemetry transmitter, arterial blood pressure catheter and IC ECG lead were implanted in 40 cynomolgus monkeys, two of which were also implanted with subcutaneous (SC) ECG leads. The data of IC ECG, heart rate (HR) and mean arterial blood pressure (MABP) were collected telemetrically for a period of 1-12 months after implantation, and measured using computer softwares., Results: the IC ECG waveforms varied greatly from those of SC ECG. There was no clearly identifiable S-T segment, and T waves were biphasic in the majority of IC ECGs. The morphology of IC ECG was diversified among animals, progressively changed in the first 2 weeks post-surgery and stabilized approximately 3 weeks post-surgery. MABP and HR were elevated after implant surgery, but recovered to the levels comparable to those of SC in approximately 1 and 4 weeks, respectively. The IC ECG values obtained during week 8 to 10 (HR=134+/-25 bpm, PR interval=87+/-13 ms, QRS interval=40+/-7 ms, and QT interval=246+/-30 ms, QTcF=318+/-28 ms) were comparable to those from SC ECG., Discussion: the IC ECG provides a clear ECG signal with values comparable to, and waveforms different from, SC recordings. The complicated surgical procedure with long substantial recovery time, high incidence of IC lead malfunction, and high costs for IC leads may limit application of the IC ECG model in safety pharmacology studies.

It is routine practice to auscultate the chest for equal bilateral breath sounds after tracheal intubation to verify that the tracheal tube tip is properly positioned. Signs of bronchial intubation also include desaturation, an increase or decrease in end-tidal carbon dioxide, and an increase in peak airway pressures. We report a case in which, during the course of endovascular coiling of a cerebral aneurysm during general anesthesia, desaturation occurred despite seemingly equal air entry on chest auscultation. The clinical picture was confused further by the presence of normal airway pressures and capnography. Incidental detection of bronchial intubation by the radiologist while performing fluoroscopy prevented any adverse sequelae to the patient and also presented an unusual method of diagnosis.

MRC CRASH is a randomised controlled trial (ISRCTN74459797) of the effect of corticosteroids on death and disability after head injury. We randomly allocated 10,008 adults with head injury and a Glasgow Coma Scale score of 14 or less, within 8 h of injury, to a 48-h infusion of corticosteroid (methylprednisolone) or placebo. Data at 6 months were obtained for 9673 (96.7%) patients. The risk of death was higher in the corticosteroid group than in the placebo group (1248 [25.7%] vs 1075 [22.3%] deaths; relative risk 1.15, 95% CI 1.07-1.24; p=0.0001), as was the risk of death or severe disability (1828 [38.1%] vs 1728 [36.3%] dead or severely disabled; 1.05, 0.99-1.10; p=0.079). There was no evidence that the effect of corticosteroids differed by injury severity or time since injury. These results lend support to our earlier conclusion that corticosteroids should not be used routinely in the treatment of head injury.

Background: Corticosteroids have been used to treat head injuries for more than 30 years. In 1997, findings of a systematic review suggested that these drugs reduce risk of death by 1-2%. The CRASH trial--a multicentre international collaboration--aimed to confirm or refute such an effect by recruiting 20000 patients. In May, 2004, the data monitoring committee disclosed the unmasked results to the steering committee, which stopped recruitment., Methods: 10008 adults with head injury and a Glasgow coma score (GCS) of 14 or less within 8 h of injury were randomly allocated 48 h infusion of corticosteroids (methylprednisolone) or placebo. Primary outcomes were death within 2 weeks of injury and death or disability at 6 months. Prespecified subgroup analyses were based on injury severity (GCS) at randomisation and on time from injury to randomisation. Analysis was by intention to treat. Effects on outcomes within 2 weeks of randomisation are presented in this report. This study is registered as an International Standard Randomised Controlled Trial, number ISRCTN74459797., Findings: Compared with placebo, the risk of death from all causes within 2 weeks was higher in the group allocated corticosteroids (1052 [21.1%] vs 893 [17.9%] deaths; relative risk 1.18 [95% CI 1.09-1.27]; p=0.0001). The relative increase in deaths due to corticosteroids did not differ by injury severity (p=0.22) or time since injury (p=0.05)., Interpretation: Our results show there is no reduction in mortality with methylprednisolone in the 2 weeks after head injury. The cause of the rise in risk of death within 2 weeks is unclear.

BslI restriction endonuclease cleaves the symmetric sequence CCN(7)GG (where N=A, C, G or T). The enzyme is composed of two subunits, alpha and beta, that form a heterotetramer (alpha(2)beta(2)) in solution. The alpha subunit is believed to be responsible for DNA recognition, while the beta subunit is thought to mediate cleavage. Here, for the first time, we provide experimental evidence that BslI binds Zn(II). Specifically, using X-ray absorption spectroscopic analysis we show that the alpha subunit of BslI contains two Zn(Cys)(4)-type zinc motifs similar to those in the DNA-binding domain of the glucocorticoid receptor. This conclusion is supported by genetic analysis of the zinc-binding motifs, whereby amino acid substitutions in the zinc finger motifs are demonstrated to abolish or impair cleavage activity. An additional putative zinc-binding motif was identified in the beta subunit, consistent with the X-ray absorption data. These data were corroborated by proton induced X-ray emission measurements showing that full BslI contains at least three fully occupied Zn sites per alpha/beta heterodimer. On the basis of these data, we propose a role for the BslI Zn motifs in protein-DNA as well as protein-protein interactions.

Transepidermal water loss (TEWL) describes the total amount of water lost through the skin, a loss that occurs constantly by passive diffusion through the epidermis. Although TEWL is a normal physiological phenomenon, if it rises too high, the skin can become dehydrated, disrupting form and function and potentially leading to infection or transepidermal passage of deleterious agents. We have validated the use of the Evaporimeter EP-2 for the accurate assessment of TEWL in the canine. We have identified a requirement for the subject to be completely still during measurements, a requirement that can be fulfilled by training. It was found that, following training of the subject, the mean TEWL value dropped, on average, 47% compared to that of untrained animals. A significant effect on TEWL of shaving the coat from the area to be measured was identified. Using the refined protocol we observed that TEWL tended to be higher in adult (2-7 y) than senior (8-11 y) dogs, suggesting that aging processes may be occurring in canine skin that impact barrier function. The implications of poorer barrier function with age could be increased susceptibility to certain skin conditions. The overall poorer skin and coat condition seen for many older dogs may also be related to an increased TEWL.

The People's Republic of China has significantly improved the general health of its people by a concerted effort in primary health care but trauma care and its prevention remains a problem. This paper provides an overview of the strengths and weaknesses of the trauma-care system in China and proposes a strategy for its future development. This includes public-health legislation, the integration of military and civilian practice to provide comprehensive care from the scene of the incident through to rehabilitation, medical audit, the introduction of postgraduate trauma-management training courses and international academic exchanges.

Nitric oxide (NO) gas is produced by various cells within the lower respiratory tract, including inflammatory and epithelial cells, and is detectable in the exhaled air of normal human subjects. We have measured exhaled NO in patients with asthma, since several cell types that are activated in asthma can produce NO after induction. NO was measured reproducibly by a slow vital capacity manoeuvre and an adapted chemiluminescence analyser. NO was detectable in exhaled air of 67 control subjects (mean peak concentration 80.2 [SE 4.1] ppb) and was significantly reduced by inhalation of the specific NO synthase inhibitor NG-monomethyl-L-arginine. 61 non-steroid-treated asthmatic subjects had significantly higher peak expired NO concentrations than controls (283 [16] ppb, p < 0.001) but 52 asthmatic patients receiving inhaled corticosteroids had levels similar to controls (101 [7] ppb). High exhaled NO concentrations in asthmatic patients may reflect induction of NO synthase, which is known to be inhibited by steroids. Measurement of exhaled NO concentrations may be clinically useful in detection and management of cytokine-mediated inflammatory lung disorders.