Mass-zero constrained dynamics for simulations based on orbital-free density functional theory.

A new algorithm for efficient and fully time-reversible integration of first-principles molecular dynamics based on orbital-free density functional theory (OFDFT) is presented. The algorithm adapts to this nontrivial case, the recently introduced Mass-Zero (MaZe) constrained dynamics. The formalism ensures that full adiabatic separation is enforced between nuclear and electronic degrees of freedom and, consequently, that the exact Born-Oppenheimer probability for the nuclei is sampled. Numerical integration of the MaZe dynamics combines standard molecular dynamics algorithms, e.g., Verlet or velocity Verlet, with the SHAKE method to impose the minimum conditions on the electronic degrees of freedom as a set of constraints. The developments presented in this work, which include a bespoke adaptation of the standard SHAKE algorithm, ensure that the quasilinear scaling of OFDFT is preserved by the new method for a broad range of kinetic and exchange-correlation functionals, including nonlocal ones. The efficiency and accuracy of the approach are demonstrated via calculations of static and dynamic properties of liquid sodium in the constant energy and constant temperature ensembles.

Active site-labeled prothrombin inhibits prothrombinase in vitro and thrombosis in vivo.

Mouse and human prothrombin (ProT) active site specifically labeled with D-Phe-Pro-Arg-CH(2)Cl (FPR-ProT) inhibited tissue factor-initiated thrombin generation in platelet-rich and platelet-poor mouse and human plasmas. FPR-prethrombin 1 (Pre 1), fragment 1 (F1), fragment 1.2 (F1.2), and FPR-thrombin produced no significant inhibition, demonstrating the requirement for all three ProT domains. Kinetics of inhibition of ProT activation by the inactive ProT(S195A) mutant were compatible with competitive inhibition as an alternate nonproductive substrate, although FPR-ProT deviated from this mechanism, implicating a more complex process. FPR-ProT exhibited ∼10-fold more potent anticoagulant activity compared with ProT(S195A) as a result of conformational changes in the ProT catalytic domain that induce a more proteinase-like conformation upon FPR labeling. Unlike ProT and ProT(S195A), the pathway of FPR-ProT cleavage by prothrombinase was redirected from meizothrombin toward formation of the FPR-prethrombin 2 (Pre 2)·F1.2 inhibitory intermediate. Localization of ProT labeled with Alexa Fluor® 660 tethered through FPR-CH(2)Cl ([AF660]FPR-ProT) during laser-induced thrombus formation in vivo in murine arterioles was examined in real time wide-field and confocal fluorescence microscopy. [AF660]FPR-ProT bound rapidly to the vessel wall at the site of injury, preceding platelet accumulation, and subsequently to the thrombus proximal, but not distal, to the vessel wall. [AF660]FPR-ProT inhibited thrombus growth, whereas [AF660]FPR-Pre 1, lacking the F1 membrane-binding domain did not bind or inhibit. Labeled F1.2 localized similarly to [AF660]FPR-ProT, indicating binding to phosphatidylserine-rich membranes, but did not inhibit thrombosis. The studies provide new insight into the mechanism of ProT activation in vivo and in vitro, and the properties of a unique exosite-directed prothrombinase inhibitor.

Non-stroke admissions to a hyperacute stroke unit.

A significant proportion of patients presenting to hyperacute stroke units (HSUs) are diagnosed with non-stroke (NS). This study aimed to assess the rate and diagnoses of NS patients admitted to a HSU and the implications for clinical service provision. Admissions to the HSU at the Southern General Hospital, Glasgow, were retrospectively assessed (March 2007-September 2007). NS patients were identified by two parallel ascertainment methods and NS diagnosis was confirmed by case-note and discharge letter review. Of 375 presentations, 116 (31%) were due to NS. NS diagnosis was more likely for local referrals than from regional hospitals (41% versus 19%; P = 0.0002). Compared with stroke/transient ischaemic attack patients, NS patients were significantly younger, more likely to have an magnetic resonance imaging (MRI) scan and had a shorter length of hospital stay. Common NS diagnoses were migraine (22%), functional neurological disorder (14%), syncope (12%) and seizure (6%). NS patients who had an MRI scan were more likely to have a length of stay ≥2 days (75% versus 53%; P = 0.03). NS makes up one-third of acute stroke-like presentations with a high frequency of neurological conditions. NS patients tend to be younger and require significant investigation. The increased use of MRI and neurological services has implications for providing a hyperacute stroke service.

Opioid-like adverse effects of tianeptine in male rats and mice.

RATIONALE: Tianeptine is a mu-opioid receptor (MOR) agonist with increasing reports of abuse in human populations. Preclinical data regarding the abuse potential and other opioid-like adverse effects of tianeptine at supratherapeutic doses are sparse. OBJECTIVES: The present study evaluated tianeptine in a rat model of abuse potential assessment and in mouse models of motor, gastrointestinal, and respiratory adverse effects. METHODS: Abuse potential was assessed in adult male Sprague-Dawley rats using an intracranial self-stimulation (ICSS) procedure to determine effects of acute and repeated tianeptine on responding for electrical brain stimulation. Male ICR mice were used to determine the effects of tianeptine in assays of locomotor behavior and gastrointestinal motility. Male Swiss-Webster mice were monitored for respiratory changes using whole-body plethysmography. RESULTS: In rats, acute tianeptine produced weak and delayed evidence for abuse-related ICSS facilitation at an intermediate dose (10 mg/kg, IP) and pronounced, naltrexone-preventable ICSS depression at a higher dose (32 mg/kg, IP). Repeated 7-day tianeptine (10 and 32 mg/kg/day, IP) produced no increase in abuse-related ICSS facilitation, only modest tolerance to ICSS depression, and no evidence of physical dependence. In mice, tianeptine produced dose-dependent, naltrexone-preventable locomotor activation. Tianeptine (100 mg/kg, SC) also significantly inhibited gastrointestinal motility and produced naloxone-reversible respiratory depression. CONCLUSIONS: Tianeptine presents as a MOR agonist with resistance to tolerance and dependence in our ICSS assay in rats, and it has lower abuse potential by this metric than many commonly abused opioids. Nonetheless, tianeptine produces MOR agonist-like acute adverse effects that include motor impairment, constipation, and respiratory depression.

Multidrug-resistant tuberculosis in Queensland, Australia: an ongoing cross-border challenge.

SETTINGp: Multidrug-resistant tuberculosis (MDR-TB) is a growing concern worldwide. In Australia, although the incidence of MDR-TB remains low, Queensland is at an increased risk due to its proximity to Papua New Guinea (PNG). OBJECTIVE: To examine the epidemiology, clinical features and outcomes of MDR-TB in Queensland, with a comparison between cross-border PNG and non-cross-border patients. DESIGN: Retrospective case series of all MDR-TB patients in Queensland between 1 January 2000 and 31 December 2014. RESULTS: Ninety-six patients were diagnosed with MDR-TB in Queensland between 2000 and 2014. The majority were cross-border PNG nationals diagnosed within the Torres Straight Protected Zone (n = 73, 76%). Cross-border patients were younger (27.4 vs. 36.3 years, P = 0.02), had spent less time in Australia before diagnosis (<1 vs. 19 months, P < 0.01), had higher rates of smear positivity (67.1% vs. 40%, P = 0.04) and were less likely to have received a second-line injectable agent (45.8% vs. 71.4%, P = 0.05). Cross-border patients had significantly lower rates of treatment success than non-cross-border patients (47.9% vs. 85.7%; P < 0.01). CONCLUSION: MDR-TB cases in Queensland are largely a result of cross-border PNG nationals, with poorer outcomes seen in this cohort. Continued strengthening of the region's TB programmes, with a focus on cross-border patients, is required.

Perfusion thresholds in acute stroke thrombolysis.

BACKGROUND AND PURPOSE: Perfusion-weighted MRI has been shown to be useful in the early identification of cerebral tissue at risk of infarction during acute ischemia. Identification of threshold perfusion measures that predict infarction may assist in the selection of patients for thrombolysis. METHODS: Mean transit time (MTT), regional cerebral blood flow (rCBF), and regional cerebral blood volume (rCBV) maps were generated in 35 acute stroke patients (17 treated with tissue plasminogen activator and 18 control patients) imaged within 6 hours from symptom onset. Day 90 outcome infarcts (T2-weighted MRI) were superimposed on acute MTT, rCBF, and rCBV maps. Perfusion-weighted MRI measures were then calculated for 2 regions: infarcted and salvaged tissue. RESULTS: MTT was prolonged by 22% in infarcted regions relative to salvaged tissue (P<0.001). rCBF was 10% lower in infarcted tissue than in salvaged regions (P<0.01). rCBV did not differ significantly between infarcted and salvaged regions. When reperfusion occurred, tissue with more severely prolonged MTT was salvaged from infarction relative to patients with persistent hypoperfusion (P<0.05). In contrast, rCBF in salvaged regions did not differ between patients with and without reperfusion. In reperfused patients, an inverse correlation (R=0.93, P<0.001) was found between time of initial MRI scan and MTT delay in salvaged tissue. CONCLUSIONS: Both increases in MTT and decreases in rCBF predict infarction. Differences in MTT also predict salvage in more severely hypoperfused tissue after reperfusion, suggesting that it is the most clinically useful quantitative perfusion measure. Perfusion thresholds for infarction need to be assessed in the context of symptom duration.

Linkage analysis of angiotensin-converting enzyme (ACE) insertion/deletion polymorphism and systemic lupus erythematosus.

Previous studies have suggested an association between systemic lupus erythematosus (SLE) and an insertion/deletion polymorphism in the angiotensin-converting enzyme gene (ACE). This polymorphism consists of a 250-bp insertion/deletion of an alu repeat in the 16th intron of the ACE gene. Individuals homozygous for the deletion have a higher level of circulating enzyme. Due to the important role of this enzyme in regulating the renin--angiotensin and kallikrein--kininogen systems, it is possible that the ACE insertion/deletion may play a role in SLE, which can include vasculitis and vascular changes. Using primers flanking the insertion/deletion site, we have examined the ACE gene in lupus patients and family members using genomic DNA obtained from the Lupus Multiplex Registry and Repository (LMRR). We were unable to detect significant linkage or genetic association between the ACE gene and SLE.

Differential development of behavioral tolerance and the subsequent hedonic effects of alcohol in AA and ANA rats.

RATIONALE: There at least two ways in which tolerance development to alcohol's behavioral effects could interact with its subsequent intake: 1) tolerance to alcohol's reward or reinforcing effects per se could lead to increased consumption, and 2) tolerance to alcohol's aversive effects could unmask alcohol's rewarding effects. These two mechanisms may differentially interact with preexisting genetic traits underlying alcoholism. OBJECTIVES: Alcohol's subjective attributes were assessed in selectively bred AA and ANA rats after the development of tolerance to alcohol's behaviorally disruptive effects on lever-press performance. METHODS: Rats were trained to press a lever under an FR30 schedule of food presentations. Group-dependent differential access to intoxicated practice, using a typical pre-post drug administration design, was utilized to promote the development of alcohol tolerance in only the group receiving intoxicated practice sessions. Subsequently, rats were trained to associate alcohol with unique place and taste stimuli in order to assess the relative changes in the approach towards, or avoidance of alcohol-related cues in each group. RESULTS: Groups of AA and ANA rats given access to intoxicated practice demonstrated tolerance development. These groups subsequently conditioned place preferences and failed to develop conditioned taste aversions to alcohol. Passive alcohol exposure in the ANA rats set the occasion for the development of a place preference and delayed taste conditioning. AA rats exposed to passive alcohol exposure failed to condition place preferences and developed rapid taste aversions. Saline control rats failed to develop tolerance or place preferences but did condition a robust alcohol-induced taste aversion. CONCLUSIONS: AA and ANA rats differ in their behavioral and pharmacokinetic response to chronic alcohol exposure. Compensatory responses interacting with approach-avoidance behaviors appear to be learned during intoxicated practice in the AA rats and during both intoxicated practice and passive exposure in the ANA rat line.

Linear free energy relationships implicate three modes of binding for fluoroaromatic inhibitors to a mutant of carbonic anhydrase II.

[figure: see text] Linear free energy relationships between binding affinity and hydrophobicity for a library of fluoroaromatic inhibitors of F131V carbonic anhydrase II (CA) implicate three modes of interaction. X-ray crystal structures suggest that F131 interacts with fluoroaromatic inhibitors, while P202, on the opposite side of the active site cleft, serves as the site of the hydrophobic contact in the case of the F131V mutant. 2-Fluorinated compounds bind more tightly, perhaps due to the field effect of the nearby fluorine on the acidity of the amide proton.

Antalarmin blockade of corticotropin releasing hormone-induced hypertension in rats.

Central administration of CRH results in endocrinological, cardiovascular, and behavioral effects that suggest stress or anxiety. Among these is a marked pressor response. Parenteral administration of CRH, however, results in hypotension. We used parenteral administration of antalarmin, a novel, small molecule CRH1 receptor antagonist, and alpha-helical CRH(9-41), a peptidic CRHR1/CRHR2 antagonist to attempt to determine the receptor mechanisms through which CRH is acting in both of these situations. Our results suggest that the hypertension produced by central CRH administration is mediated through central CRHR1 receptors, whereas the hypotension produced by parenteral CRH administration is mediated through peripheral CRHR2 receptors.

Electronic reporting to improve patient safety.

BACKGROUND: Limited data are available on the experiences of voluntary event reporting systems to improve patient safety. OBJECTIVE: Development and implementation of educational initiatives to facilitate the use of an electronic reporting system (ERS) in an academic medical center to measure the impact on knowledge of the ERS on reporting behavior and safety attitudes and to evaluate the accuracy of the information being reported. METHODS: A voluntary internal confidential electronic system for reporting safety events was implemented which involved patients and visitors. A multifaceted educational program was developed to promote safety awareness and use of the ERS system. The safety event detail reported for the calendar year 2002 was tracked and trended and central event analyses were performed for five high event clinical areas. A survey was administered to assess safety knowledge and attitudes of patient care personnel. RESULTS: 2843 safety events were entered into the ERS during 2002 with an increase during the course of the year (p = 0.055, linear trend) for all events. Nurses entered 73% of the events and physicians only 2%. 453 events (16%) were unsafe conditions or near misses and 623 (22%) were associated with patient harm. System factors were considered by the reporter as contributing to the event in only a few cases (5%). Central event analysis revealed that 39% of events had coding errors either in event classification, level of impact, or location; significant underreporting was also present. Although survey response rates were low (10.3%), responders showed a high degree of knowledge on general questions of patient safety and an increase in knowledge on use of the ERS (p = 0.0015, linear trend). CONCLUSIONS: Knowledge on the use of the reporting system and the frequency of reported events increased over the first year of the study. More work is needed to involve physicians in reporting, to improve the accuracy of submitted information, and to better prioritize, organize, and streamline event analysis.

A catalytic antibody against cocaine attenuates cocaine's cardiovascular effects in mice: a dose and time course analysis.

The murine monoclonal antibody 15A10 (mAb 15A10), elicited by a transition-state analog for cocaine hydrolysis, has previously been shown to metabolize cocaine in vitro and in vivo. The present experiments were designed to evaluate further the in vivo effectiveness of mAb 15A10 in blocking cardiovascular effects of acute cocaine administration. Balb/c mice were implanted with a femoral artery catheter utilized for mean arterial pressure (MAP) monitoring, and administered intravenous (i.v.) pretreatments of either mAb 15A10 (10, 32, 100 and 300 mg/kg) or vehicle prior to cocaine injection (100 mg/kg, i.p.). A time course analysis for mAb 15A10's effect was also conducted, for which either vehicle or 100 mg/kg mAb 15A10 was infused 1, 3, 10 and 30 days prior to cocaine treatment. During the cardiovascular recording sessions, mice were awake and freely moving within a limited area. Increases in MAP (approximately 25 mm Hg) following cocaine injection were dose-dependently attenuated by mAb 15A10. The antibody-attenuated cocaine-induced increases in MAP at 1- and 3-day pretreatment times, and reduced mortality at some of the time points studied. With 100 mg/kg antibody, plasma cocaine levels were significantly decreased early in the recording session, whereas levels of ecgonine methyl ester increased significantly. Although 10-fold greater quantities of antibody are required to observe significant effects in mouse, compared to our previous studies in rats, the present mouse model provides a convenient paradigm for investigating catalytic and non-catalytic antibodies.

Effect of infant position on breath amplitude measured by transthoracic impedance and strain gauges.

Continuous monitoring of respiration by transthoracic electrical impedance gives a signal that has certain not well understood irregularities. Among them is a change in the amplitude of the signal when there is no apparent change in the infant's tidal respiration. One factor that could hypothetically account for alterations of the impedance signal is a change in current path through the thorax secondary to a change in body position. To test this hypothesis we have studied the relationships between breath amplitude measured by transthoracic impedance, one strain gauge on the chest and one on the abdomen, and tidal volume by integrated flow in four body positions. Median breath amplitude was found to vary significantly with body position according to the measuring device. The median impedance breath amplitude increased by 27% in the supine position compared with the prone position, with no associated change in tidal volume. Differences in the strain gauge signal amplitude for these positions were not statistically significant. Correlation between breath amplitude measured by impedance changes and tidal volume was minimal (r = 0.114). These results indicate that infant position affects impedance breath amplitude independently of changes in tidal volume.

Optimal electrode location for monitoring the ECG and breathing in neonates.

Continuous monitoring of breathing in infants is commonly performed using transthoracic impedance. This method employs skin surface electrodes measuring changes in electrical impedance and relates these changes to respiratory events. Typically, two electrodes on the infant's chest monitor both the ECG and breathing. We have attempted to identify separate electrode locations that give the best signal for breathing and ECG, and a single location that optimizes both of these signals. Thirty-seven infants were studied by placing 12 electrodes on the infant's chest and abdomen, and serially sampling pairwise combinations of electrodes. The optimal signal for breathing was obtained when electrodes spanned the diaphragm. Optimal ECG signal was seen with one electrode at the right mid-clavicle and one at the xyphoid. Clinicians should be aware of these locations in order to provide the best signal available.

The stimulus properties of two common over-the-counter drug mixtures: dextromethorphan + ephedrine and dextromethorphan + diphenhydramine.

Two groups of rats were trained in a two-choice drug discrimination procedure under a fixed-ratio 10 schedule of food reinforcement. One group of rats (n=12) was trained to discriminate the presence and absence of a drug mixture containing 10 mg/kg dextromethorphan + 10 mg/kg diphenhydramine. The other group of rats (n=12) was trained to discriminate the presence and absence of another drug mixture containing 10 mg/kg dextromethorphan + 10 mg/kg ephedrine. Cross-generalization tests conducted with each of the stimulus elements demonstrated that (1) the drug mixtures were not perceived as new entities distinct from their component elements and (2) the stimulus element saliency may be a factor determining the nature of discriminative control by drug mixtures. Cross-generalization tests conducted with the psychomotor stimulants, cocaine and amphetamine, engendered complete generalization to the training cues in both groups, whereas, pentobarbital engendered predominantly saline- or default-lever responding. These data suggest a potential abuse liability for both of these common over-the-counter drug mixtures and cautions against the use of such combinations in pediatric patients.

Familial heterogeneity in infantile autism.

The incidence of autism and cognitive disability was assessed in the biological siblings of 29 autistic probands subdivided on the basis of IQ. A significant clustering of autism and nonspecific intellectual retardation was found in the siblings of severely retarded autistic probands which was not present in the siblings of our higher-functioning autistic sample. These findings suggest that there may be etiological differences in autism, depending on the degree of associated mental retardation.

Characterization of cocaine self-administration and pharmacokinetics as a function of time of day in the rat.

Two experiments examined the influence of time of day on the intravenous self-administration of cocaine and its associated pharmacokinetic profile in male Sprague-Dawley rats. In both experiments, individual rats were randomly assigned to experimental groups (n = 6/group) according to four selected times of day, 0100, 0700, 1300, and 1900 h, during which experimental procedures were conducted. In both experiments, rats were maintained under a 12 L:12 D ambient lighting cycle, with lights on at 0600 h. Training and testing was thus conducted either 1 (0700, 1900) or 7 (1300, 0100 h) hours into the light and dark phases. In Experiment 1, characteristics of cocaine self-administration across a behaviorally active dose range were assessed. Statistically significant differences were observed in the rates and patterns of self-administration across the four experimental groups, most notably characterized by an apparent shift in the dose of cocaine, which engendered peak rates of responding. Specifically, groups tested at 0100 and 1300 h appeared to exhibit enhanced sensitivity to the reinforcing properties of low-dose cocaine relative to groups tested at 0700 and 1900 h. The observed differences in apparent sensitivity of experimental subjects to low-dose cocaine were not related in any simple way to ongoing patterns of general locomotor activity, and were not accompanied by corresponding variance in the pharmacokinetic profiles of cocaine when assessed over 1 h following an intravenous infusion (1.8 mg/kg) at each of the four sampling periods noted above.

Discriminative effects of compound drug stimuli: a focus on attention.

Discrimination research has increasingly used compound stimuli emerging from drugs acting through multiple neurotransmitter systems or from injections of drug mixtures that approximate "street-wise" drug-taking behaviors. Accompanying this trend has been an interest in the role of cognitive factors in drug discrimination learning. Accounts of multidimensional drug stimuli have focused mainly on specific neuronal mechanisms, and have largely ignored the contribution of stimulus information to the perception of internal events or to the selection processes, heretofore called attention mechanisms, which may underlie the observer's idiosyncratic response to drug administration. It is argued here that research in drug discrimination may benefit from a more detailed consideration of the processes by which an observer interacts with the emergent stimulus properties of drug administration. Therapeutic intervention initiatives may critically depend on knowing the interactions between the specific attributes of the drug experience that capture the attention of the individual and that may later acquire stimulus control over complex drug-taking behaviors.

Cross-generalization of an EtOH "hangover" cue to endogenously and exogenously induced stimuli.

Twenty male Sprague-Dawley rats were trained in a two-choice food-reinforced drug discrimination task (10 min sessions) using the state-dependent interoceptive stimulus attributes of ethanol's (EtOH) delayed or rebound effects (EDE) versus "normal" basal homeostasis. Cross-generalization tests were conducted with 0.18 mg/kg naloxone injected after three days of three injections per day of either SAL or 10 mg/kg morphine. Naloxone failed to generalize to the EDE-state after chronic saline; however, the precipitated morphine withdrawal state produced complete generalization to the EDE training cue. Daily tests were conducted after 8 h photoperiod phase-shifts. An 8 h phase-advance, equivalent to a west-to-east intercontinental night-time flight in humans, produced a biphasic, graded, increase in EDE-appropriate responding, which peaked on the second day after the phase-advance and recovered by the fourth day. The 8 h phase-delays failed to engender significant EDE-appropriate responding. These data provide evidence for the subjective similarity between EtOH hangover, opiate withdrawal states, and the physiological disruption induced by circadian phase-advances.

Cumulative vs. acute dose-response procedures produce differential BAC and behavioral functions for ethanol.

The discriminative stimulus attributes of ethanol (ETOH) were characterized in rats trained to discriminate between 1.25 g/kg ETOH and saline. The ETOH generalization functions were assessed using both acute and cumulative dosing procedures. The cumulative procedures differed in the individual incremented doses used to generate the functions. Acute dosing procedures produced discriminative functions that were significantly different from cumulative dose-response curves (DRCs). Similar cumulative DRC's were generated within each cumulative dosing procedure, whereas significant differences were produced between the two dosing incremented procedures. When blood alcohol concentrations (BACs) were quantified, a cumulative testing procedure produced significantly lower BACs than acute testing procedures at every dose above the initial or starting dose. Interestingly, response rate functions did not differ within or between cumulative and acute procedures. These data may suggest that differential ETOH dosing procedures may differentially influence the behavioral choice and BAC functions in rats, and cautions against the use of cumulative procedures to assess shifts in DRCs during chronic treatments without a concomitant assessment of BACs.