Increase in Blood-Brain Barrier (BBB) Permeability Is Regulated by MMP3 via the ERK Signaling Pathway.

BACKGROUND: The blood-brain barrier (BBB) regulates the exchange of molecules between the brain and peripheral blood and is composed primarily of microvascular endothelial cells (BMVECs), which form the lining of cerebral blood vessels and are linked via tight junctions (TJs). The BBB is regulated by components of the extracellular matrix (ECM), and matrix metalloproteinase 3 (MMP3) remodels the ECM's basal lamina, which forms part of the BBB. Oxidative stress is implicated in activation of MMPs and impaired BBB. Thus, we investigated whether MMP3 modulates BBB permeability. METHODS: Experiments included in vivo assessments of isoflurane anesthesia and dye extravasation from brain in wild-type (WT) and MMP3-deficient (MMP3-KO) mice, as well as in vitro assessments of the integrity of monolayers of WT and MMP3-KO BMVECs and the expression of junction proteins. RESULTS: Compared to WT mice, measurements of isoflurane usage and anesthesia induction time were higher in MMP3-KO mice and lower in WT that had been treated with MMP3 (WT+MMP3), while anesthesia emergence times were shorter in MMP3-KO mice and longer in WT+MMP3 mice than in WT. Extravasation of systemically administered dyes was also lower in MMP3-KO mouse brains and higher in WT+MMP3 mouse brains, than in the brains of WT mice. The results from both TEER and Transwell assays indicated that MMP3 deficiency (or inhibition) increased, while MMP3 upregulation reduced barrier integrity in either BMVEC or the coculture. MMP3 deficiency also increased the abundance of TJs and VE-cadherin proteins in BMVECs, and the protein abundance declined when MMP3 activity was upregulated in BMVECs, but not when the cells were treated with an inhibitor of extracellular signal related-kinase (ERK). CONCLUSION: MMP3 increases BBB permeability following the administration of isoflurane by upregulating the ERK signaling pathway, which subsequently reduces TJ and VE-cadherin proteins in BMVECs.

Effects of Volatile Anesthetics on Postoperative Ischemic Stroke Incidence.

Background Preclinical studies suggest that volatile anesthetics decrease infarct volume and improve the outcome of ischemic stroke. This study aims to determine their effect during noncardiac surgery on postoperative ischemic stroke incidence. Methods and Results This was a retrospective cohort study of surgical patients undergoing general anesthesia at 2 tertiary care centers in Boston, MA, between October 2005 and September 2017. Exclusion criteria comprised brain death, age <18 years, cardiac surgery, and missing covariate data. The exposure was defined as median age-adjusted minimum alveolar concentration of all intraoperative measurements of desflurane, sevoflurane, and isoflurane. The primary outcome was postoperative ischemic stroke within 30 days. Among 314 932 patients, 1957 (0.6%) experienced the primary outcome. Higher doses of volatile anesthetics had a protective effect on postoperative ischemic stroke incidence (adjusted odds ratio per 1 minimum alveolar concentration increase 0.49, 95% CI, 0.40-0.59, P<0.001). In Cox proportional hazards regression, the effect was observed for 17 postoperative days (postoperative day 1: hazard ratio (HR), 0.56; 95% CI, 0.48-0.65; versus day 17: HR, 0.85; 95% CI, 0.74-0.99). Volatile anesthetics were also associated with lower stroke severity: Every 1-unit increase in minimum alveolar concentration was associated with a 0.006-unit decrease in the National Institutes of Health Stroke Scale (95% CI, -0.01 to -0.002, P=0.002). The effects were robust throughout various sensitivity analyses including adjustment for anesthesia providers as random effect. Conclusions Among patients undergoing noncardiac surgery, volatile anesthetics showed a dose-dependent protective effect on the incidence and severity of early postoperative ischemic stroke.

End-tidal to Arterial Gradients and Alveolar Deadspace for Anesthetic Agents.

BACKGROUND: According to the "three-compartment" model of ventilation-perfusion ((Equation is included in full-text article.)) inequality, increased (Equation is included in full-text article.)scatter in the lung under general anesthesia is reflected in increased alveolar deadspace fraction (VDA/VA) customarily measured using end-tidal to arterial (A-a) partial pressure gradients for carbon dioxide. A-a gradients for anesthetic agents such as isoflurane are also significant but have been shown to be inconsistent with those for carbon dioxide under the three-compartment theory. The authors hypothesized that three-compartment VDA/VA calculated using partial pressures of four inhalational agents (VDA/VAG) is different from that calculated using carbon dioxide (VDA/VACO2) measurements, but similar to predictions from multicompartment models of physiologically realistic "log-normal" (Equation is included in full-text article.)distributions. METHODS: In an observational study, inspired, end-tidal, arterial, and mixed venous partial pressures of halothane, isoflurane, sevoflurane, or desflurane were measured simultaneously with carbon dioxide in 52 cardiac surgery patients at two centers. VDA/VA was calculated from three-compartment model theory and compared for all gases. Ideal alveolar (PAG) and end-capillary partial pressure (Pc'G) of each agent, theoretically identical, were also calculated from end-tidal and arterial partial pressures adjusted for deadspace and venous admixture. RESULTS: Calculated VDA/VAG was larger (mean ± SD) for halothane (0.47 ± 0.08), isoflurane (0.55 ± 0.09), sevoflurane (0.61 ± 0.10), and desflurane (0.65 ± 0.07) than VDA/VACO2 (0.23 ± 0.07 overall), increasing with lower blood solubility (slope [Cis], -0.096 [-0.133 to -0.059], P < 0.001). There was a significant difference between calculated ideal PAG and Pc'G median [interquartile range], PAG 5.1 [3.7, 8.9] versus Pc'G 4.0[2.5, 6.2], P = 0.011, for all agents combined. The slope of the relationship to solubility was predicted by the log-normal lung model, but with a lower magnitude relative to calculated VDA/VAG. CONCLUSIONS: Alveolar deadspace for anesthetic agents is much larger than for carbon dioxide and related to blood solubility. Unlike the three-compartment model, multicompartment (Equation is included in full-text article.)scatter models explain this from physiologically realistic gas uptake distributions, but suggest a residual factor other than solubility, potentially diffusion limitation, contributes to deadspace.

Phenylpiperidine opioid effects on isoflurane minimum alveolar concentration in cats.

Different structurally related phenylpiperidine opioids exhibit different isoflurane-sparing effects in cats. Because minimum alveolar concentration (MAC) in cats is affected only by very high plasma concentrations of some phenylpiperidine opioids, we hypothesized these effects are caused by actions on nonopioid receptors. Using a prospective, randomized, crossover design, six cats were anesthetized with isoflurane, intubated, ventilated, and instrumented. Isoflurane MAC was measured in triplicate using a tail-clamp and bracketing technique. A computer-controlled intravenous infusion using prior pharmacokinetic models targeted plasma concentrations of 60 ng/ml fentanyl, 10 ng/ml sufentanil, or 500 ng/ml alfentanil, and isoflurane MAC was measured in duplicate. Next, naltrexone 0.6 mg/kg was administered to cats hourly during the opioid infusion, and isoflurane MAC was measured in duplicate. Blood was collected during MAC determinations to measure opioid concentrations. Responses were analyzed using repeated measures ANOVA with significance at p < .05. Alfentanil and sufentanil decreased isoflurane MAC by 16.4% and 6.4%, respectively, and these effects were completely reversed by naltrexone. Fentanyl had no significant effect on isoflurane MAC. Alfentanil and sufentanil modestly reduce isoflurane MAC via agonist effects on opioid receptors. However, these effects are too small to justify clinical use of phenylpiperidine opioids as single agents to reduce MAC in cats.

Minimum anesthetic concentration of isoflurane and sparing effect of midazolam in Quaker parrots (Myiopsitta monachus).

OBJECTIVE: To determine the effects of midazolam on the minimum anesthetic concentration (MAC) reduction of end-tidal isoflurane concentration (Fe'Iso) measured using an electrical stimulus in Quaker parrots (Myiopsitta monachus). STUDY DESIGN: Randomized crossover experimental study. ANIMALS: A group of six adult Quaker parrots, weighing 98-124 g. METHODS: Birds were anesthetized with isoflurane in oxygen delivered by mask, then tracheally intubated and mechanically ventilated. Three treatments were applied with a 4 day interval between anesthetic events. Each anesthetized bird was administered midazolam (1 mg kg-1; treatment MID1), midazolam (2 mg kg-1; treatment MID2) or electrolyte solution (control) intramuscularly. The treatments were administered using a replicated Latin square design and the observers were blinded. Based on a pilot bird, the starting Fe'Iso was 1.8%. After equilibration for 10 minutes, a supramaximal stimulus was delivered using an electrical current (20 V and 50 Hz for 10 ms) and birds were observed for non-reflex movement. The Fe'Iso was titrated by 0.1% until a crossover event was observed. The MAC was estimated using logistic regression. RESULTS: The MAC of isoflurane (MACISO) was estimated at 2.52% [95% confidence interval (CI), 2.19-2.85] with a range of 1.85-2.65%. MACISO in MID1 was 2.04% (95% CI, 1.71-2.37) and in MID2 was 1.81% (95% CI, 1.48-2.14); reductions in MACISO from control of 19% (p = 0.001) and 28% (p < 0.001), respectively. Heart rate, temperature, sex and anesthetic time were not different among treatments. CONCLUSIONS: Midazolam (1-2 mg kg-1) intramuscularly resulted in a significant isoflurane-sparing effect in response to a noxious stimulus in Quaker parrots without observable adverse effects. CLINICAL RELEVANCE: Midazolam can be used as part of a balanced anesthetic approach using isoflurane in Quaker parrots, and potentially in other psittacine species.

Plasma dopamine concentrations following dopamine infusion to isoflurane-anesthetized New Zealand White rabbits.

OBJECTIVE: To determine the pharmacokinetics of dopamine following a short infusion in isoflurane-anesthetized rabbits. STUDY DESIGN: Prospective, descriptive pharmacokinetic study. ANIMALS: A group of six adult female New Zealand White rabbits weighing 4.4 ± 0.2 kg. METHODS: Rabbits were anesthetized with isoflurane in oxygen and maintained at 1.2 × minimum alveolar concentration of isoflurane (2.3% atmosphere). Dopamine (30 µg kg-1 minute-1) was infused for 10 minutes. Arterial blood was sampled prior, during and following the infusion at various intervals for 1 hour. RESULTS: A one-compartment model with baseline concentration best fitted the time-plasma dopamine concentration data. Estimated typical population value (interindividual variability) for volume of distribution and clearance were 10.3 (232%) L kg-1 and 9.9 (508%) L minute-1 kg-1, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: There was a large degree of interindividual variation in the disposition of dopamine. The large volume of distribution and high metabolic clearance rate reported for dopamine in this study likely explains the lack of clinical efficacy of dopamine in rabbits at doses up to 30 µg kg-1 minute-1.

Effects of midazolam and nitrous oxide on the minimum anesthetic concentration of isoflurane in the ball python (Python regius).

OBJECTIVE: To evaluate the effects of midazolam and nitrous oxide (N2O) on the minimum anesthetic concentration of isoflurane (MACISO) in ball pythons. STUDY DESIGN: Prospective, crossover, randomized, semi-blinded study. ANIMALS: A total of nine healthy adult female ball pythons (Python regius) weighing 2.76 ± 0.73 kg. METHODS: In each snake, three protocols were evaluated with 2 week washouts: treatment MID-O2, midazolam (1 mg kg-1) administered intramuscularly (IM) and anesthesia induced with isoflurane-oxygen; treatment SAL-O2, saline (0.2 mL kg-1) IM and anesthesia with isoflurane-oxygen; and treatment SAL-N2O, saline IM and anesthesia with isoflurane and 50% nitrous oxide (N2O):50% oxygen. In each treatment, isoflurane was administered by face mask immediately after premedication. Snakes were endotracheally intubated and inspired and end-tidal isoflurane concentrations were monitored. The study design followed a standard bracketing technique, and the MACISO was determined using logistic regression. Electrical stimulation using a Grass stimulator connected to the base of the tail (50 V, 50 Hz, 6.5 ms pulse-1) was used as the supramaximal stimulus. Blood-gas analysis was performed on cardiac blood collected immediately following intubation and after the last stimulation. Blood-gas variables were compared over time and between treatments using linear mixed models. RESULTS: MACISO at a body temperature of 30.1 ± 0.4 °C was 1.11% (95% confidence interval, 0.94-1.28%) in SAL-O2 and was significantly decreased to 0.48% (0.29-0.67%) in MID-O2 (p < 0.001) and to 0.92% (0.74-1.09%) in SAL-N2O (p = 0.016). PO2 was significantly lower in MID-O2 and SAL-N2O than in SAL-O2. CONCLUSIONS AND CLINICAL RELEVANCE: Midazolam significantly decreased the MACISO by 57% in ball pythons, whereas addition of N2O resulted in a modest, although significant, decrease (17%). MACISO in ball pythons was lower than those previously reported in reptiles.

Most Hemodynamically Stable Method for Change From High to Low Anesthesia Flow: A Randomized Controlled Trial Comparing State Entropy, High Fresh Gas Flow for 10 Minutes, and 0.8 Ratio of End-Expired Agent Concentration to Inspired Agent Concentration.

This study was undertaken to determine the most hemodynamically stable method to low-flow anesthesia (LFA) between 10-minute administration of high fresh gas flow, 0.8 equilibration ratio (Fe/Fi), and state entropy (SE) between 40 and 60, a marker for adequate depth of anesthesia. Change from high fresh gas flow to LFA was done in 3 groups of 30 patients each: group T (time): 10 minutes; group R (ratio): Fe/Fi = 0.8, and group SE: SE = 40 to 50. A decrease in mean blood pressure or heart rate was treated with ephedrine or atropine, with study termination at more than 2 boluses of either. In group SE, no patient required ephedrine or atropine. The requirement for ephedrine was statistically higher in groups R and T than group SE. Atropine requirement was statistically higher in group R vs groups T and SE. In group R, the mean (SD) time to LFA was 43.9 (20.37) minutes, and in group SE was 151.9 (74.4) seconds. Hypotension or bradycardia did not occur when LFA was started at SE of 40 to 50 after anesthesia induction compared with LFA at 10 minutes, which caused hypotension, and Fe/Fi of 0.8, which caused hypotension and bradycardia.

Effect of oral administration of gabapentin on the minimum alveolar concentration of isoflurane in dogs.

OBJECTIVE: To determine the effect of oral administration of gabapentin (20 mg/kg) on the minimum alveolar concentration (MAC) of isoflurane in dogs. ANIMALS: 6 healthy adult dogs (3 males and 3 females with a mean ± SD body weight of 24.8 ± 1.3 kg). PROCEDURES: Each dog was anesthetized twice. Dogs were initially assigned to 1 of 2 treatments (gabapentin [20 mg/kg, PO] followed 2 hours later by anesthesia maintained with isoflurane or anesthesia maintained with isoflurane alone). A minimum of 7 days later, dogs received the other treatment. The MAC of isoflurane was determined by use of an iterative bracketing technique with stimulating electrodes placed in the maxillary buccal mucosa. Hemodynamic variables and vital parameters were recorded at the lowest end-tidal isoflurane concentration at which dogs did not respond to the stimulus. Effect of treatment on outcome variables was analyzed by use of a paired t test. RESULTS: Mean ± SD MAC of isoflurane was significantly lower when dogs received gabapentin and isoflurane (0.71 ± 0.12%) than when dogs received isoflurane alone (0.91 ± 0.26%). Mean reduction in MAC of isoflurane was 20 ± 14%. Hemodynamic variables did not differ significantly between treatments. Mean time to extubation was significantly less when dogs received gabapentin and isoflurane (6 ± 4 minutes) than when dogs received isoflurane alone (23 ± 15 minutes). CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of gabapentin 2 hours before anesthesia maintained with isoflurane had a MAC-sparing effect with no effect on hemodynamic variables or vital parameters of dogs.

Effect of α2-adrenoceptor antagonism on the minimum alveolar concentration of isoflurane in cats.

OBJECTIVE: To characterize the effect of α2-adrenoceptor antagonism on the minimum alveolar concentration of isoflurane (MACISO) in cats. STUDY DESIGN: Prospective experimental study. ANIMALS: A group of five healthy adult male neutered cats. METHODS: Cats were anesthetized with isoflurane in oxygen and instrumented. MACISO was determined in duplicate in five cats, before and during administration of atipamezole (250 µg kg-1 followed by 250 µg kg-1 hour-1) using the bracketing technique and tail clamping. Estimates of MACISO obtained before and during administration of atipamezole were compared using a two-tailed paired t test. RESULTS: MACISO during atipamezole administration (mean ± standard deviation 2.73% ± 0.07%) was significantly larger than before atipamezole administration (1.95% ± 0.13%; p < 0.0001). CONCLUSION AND CLINICAL RELEVANCE: The role of α2-adrenoceptors in inhaled anesthetic-induced immobility may be larger than previously thought. Antagonism of an α2-adrenoceptor agonist during inhalation anesthesia may result in an increase in MAC disproportionate to the MAC reduction induced by the agonist.

Effect of heart rate on the pharmacokinetics of fentanyl in dogs anesthetized with isoflurane and hydromorphone.

OBJECTIVE: To compare the pharmacokinetics of fentanyl at lower (LHR) or higher heart rate (HHR) in dogs anesthetized with isoflurane. STUDY DESIGN: Prospective, randomized, crossover controlled trial. ANIMALS: A group of six healthy 13-month-old male Beagle dogs weighing 9.9 ± 0.7 kg (mean ± standard deviation). METHODS: Dogs were allocated to two treatments: LHR (HR: 45-75 beats minute-1) and HHR (HR: 100-130 beats minute-1). Anesthesia was maintained with isoflurane and hydromorphone (0.1 mg kg-1 followed by 0.02-0.10 mg kg-1 hour-1) for both treatments. Glycopyrrolate was administered in HHR to maintain HR within the desired range. Afterwards, fentanyl (20 µg kg-1) was intravenously administered over 5 minutes. Arterial blood samples were collected for plasma fentanyl concentration measurement by liquid chromatography/mass spectrometry. The pharmacokinetics of fentanyl were compared between treatments and the differences were considered significant at p < 0.05. RESULTS: A three-compartment model best fitted the changes in plasma fentanyl concentration. Clearance (CL; mL minute-1 kg-1) was 33.2 (24.0-48.0) and 61.3 (44.5-72.7), maximum concentration (ng mL-1) 33.6 (23.4-36.6) and 20.0 (16.7-28.0), apparent volume of the rapid peripheral compartment (mL kg-1) 436 (352-723) and 925 (499-1887), apparent volume at steady state (mL kg-1) 4064 (3453-6546) and 7195 (5077-8601), cardiac index (CI; mL minute-1 m-2) 2.83 (1.98-3.67) and 4.91 (3.22-6.09) and HR (beats minute-1) 68 (49-72) and 120 (102-129) for LHR and HHR, respectively, with significant differences between treatments. Significant correlations (0.92 and 0.90) were found between CI and CL, and between HR and CL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The increase in HR and the resultant improvement in cardiac output increased fentanyl CL and volume of distribution, which resulted in a decrease in plasma fentanyl concentration in isoflurane-anesthetized dogs.

Sex effects on behavioral markers of emergence from propofol and isoflurane anesthesia in rats.

Clinical studies have demonstrated sex-related differences in recovery from surgical anesthesia. This study aimed to characterize the emergence pattern following two anesthesia regimens in both sexes of rats. We considered six different markers of emergence from anesthesia: sigh, eye blinking, forelimb movement, mastication, neck extension, and recovery of the righting reflex (RORR). Spontaneous motor activity 24 h after the anesthesia induction was also examined. Our results showed that the rank order of the emergence latency after intraperitoneal propofol, PRO, exposure was forelimb movement < sigh < blink < mastication < neck extension < RORR, while after inhaled isoflurane, ISO, anesthesia the sequence was changed as sigh < blink < mastication < forelimb movement < neck extension < RORR in both male and female rats. Moreover, the latency to emergence after PRO in female rats was significantly higher than male rats, although following ISO there was no difference between the sexes (P < 0.001; P > 0.05, respectively). Open-field testing revealed no difference in PRO and ISO spontaneous locomotor activity due to drug administration (P > 0.05). These two anesthetics presented different emergence sequences. Although clinical data suggests that females arouse faster than males from anesthesia with propofol, our intraperitoneal technique in a rodent model had the opposite effect. Pharmacokinetic analysis demonstrated increased absorption of injected propofol for the female rats in our study, emphasizing the role of sexual dimorphism in drug distribution in rodents. Despite these pharmacokinetic differences, the pharmacodynamic effects of the drugs were remarkably consistent among both sexes through emergence.

Enantioselective capillary electrophoresis for pharmacokinetic analysis of methadone and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine in equines anesthetized with ketamine and isoflurane.

An enantioselective assay for the determination of methadone and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine in equine plasma based on capillary electrophoresis with highly sulfated γ-cyclodextrin as chiral selector and electrokinetic analyte injection is described. The assay is based on liquid/liquid extraction of the analytes at alkaline pH from 0.1 mL plasma followed by electrokinetic sample injection of the analytes from the extract across a buffer plug without chiral selector. Separation occurs cationically at normal polarity in a pH 3 phosphate buffer containing 0.16% (w/v) of highly sulfated γ-cyclodextrin. The developed assay is precise (intra- and interday RSD < 4% and < 7%, respectively), is capable to determine enantiomer levels of methadone and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine in plasma down to 2.5 ng/mL, and was successfully applied to monitor enantiomer drug and metabolite levels in plasma of a pony that was anesthetized with racemic ketamine and isoflurane and received a bolus of racemic methadone and a bolus followed by constant rate infusion of racemic methadone. The data suggest that the assay is well suited for pharmacokinetic purposes.

Effects of transdermal fentanyl solution application and subsequent naloxone hydrochloride administration on minimum alveolar concentration of isoflurane in dogs.

OBJECTIVE To assess the isoflurane-sparing effect of a transdermal formulation of fentanyl solution (TFS) and subsequent naloxone administration in dogs. DESIGN Experiment. ANIMALS 6 healthy mixed-breed dogs. PROCEDURES Minimum alveolar concentration (MAC) of isoflurane was determined in each dog with a tail clamp method (baseline). Two weeks later, dogs were treated with TFS (2.7 mg/kg [1.23 mg/lb]), and the MAC of isoflurane was determined 4 and 24 hours later. After the 4-hour MAC assessment, saline (0.9% NaCl) solution was immediately administered IV and MAC was reassessed. After the 24-hour MAC assessment, naloxone hydrochloride (0.02 mg/kg [0.01 mg/lb], IV) was immediately administered and MAC was reassessed. Heart rate, respiratory rate, arterial blood pressure, end-tidal partial pressure of CO2, and oxygen saturation as measured by pulse oximetry were recorded for each MAC assessment. RESULTS Mean ± SD MAC of isoflurane at 4 and 24 hours after TFS application was 45.4 ± 4.0% and 45.5 ± 4.5% lower than at baseline, respectively. Following naloxone administration, only a minimal reduction in MAC was identified (mean percentage decrease from baseline of 13.1 ± 2.2%, compared with 43.8 ± 5.6% for saline solution). Mean heart rate was significantly higher after naloxone administration (113.2 ± 22.2 beats/min) than after saline solution administration (76.7 ± 20.0 beats/min). No significant differences in other variables were identified among treatments. CONCLUSIONS AND CLINICAL RELEVANCE The isoflurane-sparing effects of TFS in healthy dogs were consistent and sustained between 4 and 24 hours after application, and these effects should be taken into consideration when anesthetizing or reanesthetizing TFS-treated dogs.

Matrix-Assisted Laser Desorption Ionization Imaging Mass Spectrometry of Drug Distribution in Mouse Brain Tissue by High-Resolution Time-of-Flight Mass Spectrometry.

Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry provides the opportunity to visualize the distributions of drugs and metabolites in tissue specimens without requiring radioisotopes, as are used for whole-body autoradiography. However, the analysis of low-molecular-weight compounds is often difficult using the common reflectron-type MALDI time-of-flight mass spectrometers. Insufficient mass resolving power causes overlapping of the target drug peak with matrix compound or surface contaminant peaks. To solve this issue, we describe the procedure for imaging mass spectrometry using a high-mass-resolution mass spectrometer that can separate isobaric peaks.

Genomics and Proteomic Techniques.

Although general anesthesia induced by inhaled anesthetics produces definitive phenotypes (e.g., loss of mobility, amnesia, analgesia), the underlying targets of these drugs are still not clear. Genomics and proteomic techniques are discussed for measurement of global transcriptional and translational changes after inhaled anesthetic exposures. The current discussion focuses primarily on the genomic and proteomic technical methodology. We also include a discussion of network and pathway analyses for data interpretation after identification of the targets.

Genetic variability affects absolute and relative potencies and kinetics of the anesthetics isoflurane and sevoflurane in Drosophila melanogaster.

Genetic variability affects the response to numerous xenobiotics but its role in the clinically-observed irregular responses to general anesthetics remains uncertain. To investigate the pharmacogenetics of volatile general anesthetics (VGAs), we developed a Serial Anesthesia Array apparatus to expose multiple Drosophila melanogaster samples to VGAs and behavioral assays to determine pharmacokinetic and pharmacodynamic properties of VGAs. We studied the VGAs isoflurane and sevoflurane in four wild type strains from the Drosophila Genetic Reference Panel, two commonly used laboratory strains (Canton S and w 1118 ), and a mutant in Complex I of the mitochondrial electron transport chain (ND23 60114 ). In all seven strains, isoflurane was more potent than sevoflurane, as predicted by their relative lipid solubilities, and emergence from isoflurane was slower than from sevoflurane, reproducing cardinal pharmacokinetic and pharmacodynamic properties in mammals. In addition, ND23 60114 flies were more sensitive to both agents, as observed in worms, mice, and humans carrying Complex I mutations. Moreover, we found substantial variability among the fly strains both in absolute and in relative pharmacokinetic and pharmacodynamic profiles of isoflurane and sevoflurane. These data indicate that naturally occurring genetic variations measurably influence cardinal pharmacologic properties of VGAs and that flies can be used to identify relevant genetic variations.

EFFECTS OF TRAMADOL ON THE MINIMUM ANESTHETIC CONCENTRATION OF ISOFLURANE IN WHITE-EYED PARAKEETS (PSITTACARA LEUCOPHTHALMUS).

The aim of this study was to determine the minimum anesthetic concentration (MAC) of isoflurane, and to investigate if tramadol changes the isoflurane MAC in white-eyed parakeets (Psittacara leucophthalmus). Ten adult birds weighing 157 ± 9 g were anesthetized with isoflurane in oxygen under mechanical ventilation. Isoflurane concentration for the first bird was adjusted to 2.2%, and after 15 min an electrical stimulus was applied in the thigh area to observe the response (movement or nonmovement). Isoflurane concentration for the subsequent bird was increased by 10% if the previous bird moved, or decreased by 10% if the previous bird did not move. This procedure was performed serially until at least four sequential crossover events were detected. A crossover event was defined as a sequence of two birds with different responses (positive or negative) to the electrical stimulus. Isoflurane MAC was calculated as the mean isoflurane concentration value at the crossover events. After 1 wk, the same birds were reanesthetized with isoflurane and MAC was determined at 15 and 30 min after intramuscular administration of 10 mg/kg of tramadol using the same method. A paired t-test (P < 0.05%) was used to detect significant differences for MAC between treatments. Isoflurane MAC in this population of white-eyed parakeets was 2.47 ± 0.09%. Isoflurane MAC values 15 and 30 min after tramadol administration were indistinguishable from each other (pooled value was 2.50 ± 0.18%); they were also indistinguishable from isoflurane MAC without tramadol. The isoflurane MAC value in white-eyed parakeets is higher than reported for other bird species. Tramadol (10 mg/kg, i.m.) does not change isoflurane MAC in these birds.

Temporal accumulation and localization of isoflurane in the C57BL/6 mouse and assessment of its potential contamination in 19 F MRI with perfluoro-crown-ether-labeled cardiac progenitor cells at 9.4 Tesla.

PURPOSE: To assess the uptake, accumulation, temporal stability, and spatial localization of isoflurane (ISO) in the C57BL/6 mouse, and to identify its potential interference with the detection of labeled cardiac progenitor cells using 19 F MRI/MR spectroscopy (MRS). MATERIALS AND METHODS: Objectives are demonstrated using (a) in vitro ISO tests, (b) in vivo temporal accumulation/spatial localization C57BL/6 studies (n = 3), and (c) through injections of perfluoro-crown-ether (PFCE) labeled cardiac progenitor cells into femoral muscle areas of the murine hindlimb post-mortem (n = 1) using 1 H/19 F MRI/MRS at 9.4 Tesla. Data were acquired using double-gated spoiled gradient echo images and pulse-acquire spectra. For the in vivo study, the temporal stability of ISO resonances was quantified using coefficient of variability (CV) (5 min) estimates. RESULTS: Two ISO resonances were observed in vivo that correspond to the -CF3 and -OCHF2 moieties. CV values ranged between 3.2 and 6.4% (-CF3 ) and 6.4 and 11.2% (-OCHF2 ). Reductions of the ISO dose (2.0 to 1.7%) at 80 min postinduction had insignificant effects on ISO signals (P = 0.23; P = 0.71). PFCE-labeled cells exhibited a resonance at -16.25 ppm in vitro that did not overlap with the ISO resonances, a finding that is confirmed with MRS post-mortem using injected, labeled cells. Based on 19 F MRI, similar in vivo/post-mortem ISO compartmentalization was also confirmed in peripheral and thoracic skeletal muscles. CONCLUSION: Significant ISO accumulation was observed by 19 F MRS in vivo with temporally stable signals over 90 min postinduction. ISO effects on PFCE labels are anticipated to be minimal but may be more prominent for perfluoropolyether or perfluorooctyl bromide labels. LEVEL OF EVIDENCE: 1 Technical Efficacy: Stage 1 J. MAGN. RESON. IMAGING 2017;45:1659-1667.

Awakening arterial blood and end-tidal concentrations of isoflurane in female surgical patients.

Delayed extubation occurs after isoflurane anesthesia, especially following prolonged surgical duration. We aimed to determine the arterial blood concentrations of isoflurane and the correlation with end-tidal concentrations for predicting emergence from general anesthesia.Thirty-four American Society of Anesthesiologists physical status class I-II gynecologic patients were included. General anesthesia was maintained with a fixed 2% inspiratory isoflurane in 6 L/minute oxygen, which was discontinued after surgery. One milliliter of arterial blood was obtained for the determination of isoflurane concentration by gas chromatography at 20 and 10 minutes before and 0, 5, 10, 15, and 20 minutes after discontinuation, in addition to the time of eye opening to verbal command, defined as awakening. Inspiratory and end-tidal concentrations were simultaneously detected by an infrared analyzer.The mean awakening arterial blood concentration of isoflurane was 0.20%, which was lower than the simultaneous end-tidal concentration 0.23%. The differences between arterial and end-tidal concentrations during emergence fell into an acceptable range (±1.96 standard deviation). After receiving a mean time of 108-minute general anesthesia, the time to eye opening after discontinuing isoflurane was 18.5 minutes (range 11-30, median 18 minutes), without statistical significance with anesthesia duration (P = 0.078) and body mass index (P = 0.170).We demonstrated the awakening arterial blood concentration of isoflurane in female patients as 0.20%. With well-assisted ventilation, the end-tidal concentration could be an indicator for the arterial blood concentration to predict emergence from shorter duration of isoflurane anesthesia.