Different Concentrations Of Isoflurane Research Articles

Isoflurane Preconditioning Alleviates Hypoxia/Reoxygenation-Induced Cardiomyocyte Injury by Inhibiting miR-195-3p Expression.

To investigate the role of microRNA-195-3p (miR-195-3p) in hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury. AC16 human cardiomyocyte cells were cultured and pretreated with different concentrations of isoflurane (ISO) (1%, 2%, and 3%), followed by 6h each of hypoxia and reoxygenation to construct H/R cell models. The optimum ISO concentration was assessed based on the cell viability. miR-195-3p expression was regulated by in vitro celltransfection. Cell viability was determined by MTT assay, and apoptosis was evaluated by flow cytometry. The levels of myocardial injury and inflammation were determined by enzyme-linked immunosorbent assay. Compared with the control group, the cell viability of the H/R group had significantly decreased and that of ISO pretreatment had increased in a dose-dependent manner. Therefore, we selected a 2% ISO concentration for pretreatment. MiR-195-3p expression had significantly increased in the H/R group and decreased after 2% ISO pretreatment. Additionally, the number of apoptotic cells and the levels of lactate dehydrogenase, creatine kinase-myoglobin binding, cardiactroponin I, interleukin (IL)-1β, IL-6, and tumor necrosis factor-α had increased significantly. ISO preconditioning inhibited H/R-induced AC16 cell damage, whereas miR-195-3p overexpression reversed the protective effects of ISO on cardiomyocytes. The expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) was reduced in the H/R-induced AC16 cells, and PTEN is a downstream target gene of miR-195-3p. Preconditioning with 2% ISO plays a protective role in H/R-induced AC16 cell damage by inhibiting miR-195-3p expression.

Isoflurane Promotes Cell Proliferation, Invasion, and Migration by Regulating BACH1 and miR-375 in Prostate Cancer Cells In Vitro.

The aim of this study was to investigate the mechanism of isoflurane in proliferation, invasion, and migration in prostate cancer (PC) cells in vitro by regulating BACH1 and miR-375. The effect of different concentrations of isoflurane (0%, 0.5%, 1%, and 2%) on PC cell proliferation (PC3 and 22RV1) was measured. After PC cells and normal human prostate stromal immortalized WPMY-1 cells were treated with isoflurane, BACH1 and miR-375 expression was measured. Subsequently, PC3 and 22RV1 cells underwent gain- and loss-of-function assays with or without 4-h 2% isoflurane pretreatment. The levels of miR-375, BACH1, and PTEN were assessed. The binding of BACH1 to miR-375 promoter was detected by ChIP assay. Dual-luciferase reporter assay detected the targeting relationship of miR-375 with BACH1 and PTEN. Isoflurane promoted PC3 and 22RV1 cell proliferation. In addition, isoflurane elevated the levels of BACH1 and miR-375 in a dosage-dependent manner in PC cells. Transfection with miR-375 inhibitor or sh-BACH1 repressed PC cell proliferation, invasion, and migration, while exposure to 2% isoflurane for 4 h before transfection counteracted the inhibitory effects of sh-BACH1 or miR-375 inhibitor on PC cells. PTEN expression was suppressed after 2% isoflurane treatment, but the transfection with miR-375 inhibitor partly abrogated this suppressive effect in PC cells. Moreover, BACH1 bound to miR-375 and miR-375 negatively targeted PTEN. miR-375 mimic could partially reverse the inhibitory effects of sh-BACH1 on the proliferation, invasion, and migration of isoflurane-treated PC cells. Isoflurane facilitated PC cell proliferation, migration, and invasion by activating BACH1 to upregulate miR-375.

Sevoflurane and isoflurane inhibit KCl-induced, Rho kinase-mediated, and PI3K-participated vasoconstriction in aged diabetic rat aortas

BackgroundThe mechanism of volatile anesthetics on vascular smooth muscle (VSM) contraction in the setting of diabetes mellitus (DM) remains unclear. The current study was designed to determine the effects of sevoflurane (SEVO) and isoflurane (ISO) on phosphoinositide 3-kinase (PI3K) and Rho kinase (ROCK) mediated KCl-induced vasoconstriction in aged type 2 diabetic rats.MethodsKCl-induced (60 mM) contractions were examined in endothelium-denuded aortic rings from aged T2DM Otsuka Long-Evans Tokushima Fatty (OLETF) rats (65–70 weeks old), control age-matched nondiabetic Long-Evans Tokushima Otsuka (LETO) rats and young Wistar rats (6–8 weeks old). The effects of SEVO or ISO (1–3 minimum alveolar concentration, MAC) on KCl-induced vasoconstriction, as well as those of LY294002 (PI3K inhibitor) and Y27632 (ROCK inhibitor) were measured in aortic rings from the three groups using an isometric force transducer.ResultsKCl induced rapid and continuous contraction of aortic smooth muscle in the three groups, and the contraction was more obvious in OLETF rats.SEVO and ISO inhibited KCl-induced vasoconstriction in a concentration-dependent manner and were suppressed by LY294002 (10 µM) and Y27632 (1 µM). SEVO had a stronger inhibitory effect on the aortas of young Wistar rats than ISO, especially at 2 MAC and 3 MAC (P < 0.05). In aged rats, the inhibitory effect of ISO was stronger than that of SEVO, especially OLETF rats. There was no significant difference in the effects of different concentrations of ISO on arterial contraction among the three groups (P > 0.05). The effects of 1 MAC SEVO on Wistar rats and 3 MAC SEVO on OLETF rats, however, were noticeably and significantly different (P < 0.05).Compared with the control condition, LY294002 and Y27632 had the most noticeable effect on the KCl-induced contraction of aortic rings in OLETF rats (P < 0.01).ConclusionSEVO (3 MAC), ISO (1, 2, 3 MAC), LY294002 and Y27632 have more significant inhibitory effect on the contraction of vascular smooth muscle in aged T2MD rats. The mechanism of SEVO and ISO in vascular tension in T2DM is partly due to changes in PI3K and/or Rho kinase activity.

The Effect of General Anaesthesia on Circadian Rhythms in Behaviour and Clock Gene Expression of Drosophila melanogaster.

General anaesthesia (GA) is implicated as a cause of postoperative sleep disruption and fatigue with part of the disturbance being attributed to a shift of the circadian clock. In this study, Drosophila melanogaster was used as a model to determine how Isoflurane affects the circadian clock at the behavioural and molecular levels. We measured the response of the clock at both of these levels caused by different durations and different concentrations of Isoflurane at circadian time 4 (CT4). Once characterized, we held the duration and concentration constants (at 2% in air for 6 h) and calculated the phase responses over the entire circadian cycle in both activity and period expression. Phase advances in behaviour were observed during the subjective day, whereas phase delays were associated with subjective night time GA interventions. The corresponding pattern of gene expression preceded the behavioural pattern by approximately four hours. We discuss the implications of this effect for clinical and research practice.

Short or moderate-time exposure to the inhalational anesthetics isoflurane and sevoflurane does not alter the marble-burying behavior in mice

Several studies suggest the involvement of glutamatergic neurotransmission in obsessive-compulsive disorder (OCD). Some NMDA glutamatergic receptor antagonists, such as the general anesthetic ketamine, have shown anti-OCD effects in preclinical and clinical studies. Therefore, we investigated whether the inhalational anesthetics isoflurane and sevoflurane, which are general anesthetics acting as NMDA receptor antagonists, would induce the same effects. To test our hypothesis, adult male Swiss mice were exposed to different concentrations of isoflurane (0.5, 1.5 or 3 %) or sevoflurane (0.8, 2.5 or 4 %) for 20 min (short-time exposure) or 1 h (moderate-time exposure) and submitted to the open field test (OFT) and the marble-burying test (MBT) in the same day (acute effect) or 7 days (long-lasting effect) after anesthetics administration. We found that single short or moderate-time exposure to isoflurane or sevoflurane, at sub-anesthetic or anesthetic concentrations, did not affect marble-burying behavior acutely or even 7 days after their administration. The same treatment schedules with isoflurane or sevoflurane did not impair total distance travelled in the OFT. A single moderate-time exposure to isoflurane (3 %) reduced, acutely, the central exploration of the open field, suggesting an anxiogenic-like effect of isoflurane in mice. Our results suggest that isoflurane and sevoflurane may not be promising anti-compulsive drugs.

Anesthesia differentially modulates neuronal and vascular contributions to the BOLD signal

Most studies involving BOLD fMRI in basic neuroscience research are conducted with anesthetized animals. This study investigates neural and hemodynamic activity through a combination of experiments comprising BOLD fMRI, optical calcium recordings and ASL in vivo. Patch clamp experiments of neurons were conducted to evaluate electrophysiological correlates of neural activity in vitro. Various anesthetic conditions embracing numerous anesthetic depths evoked by different concentrations of isoflurane (ISO) and different degrees of hypercapnia under a constant stimulus were investigated. We observed that different anesthetic conditions had major impact on the results obtained, particularly that anesthesia could cause a massive divergence of different experimental modalities.In ventilated animals, robust BOLD responses were detectable even with relatively deep anesthesia, while in non-ventilated animals, BOLD responses were not detectable under these conditions. This was most likely due to hypercapnia caused by respiratory depression, as in ventilated animals administered CO2 had the same effect. This observation agreed with measurements of perfusion, which showed that inhaled CO2 increased perfusion significantly, while ISO did not.In optical calcium measurements, higher concentrations of ISO decreased spontaneous neural activity, but not stimulus-evoked responses. This observation was explained by a generally lower excitability of neurons under ISO, which suppressed spontaneous activity, and consequently left more neurons available to fire synchronously in response to a stimulus. Interpreting this phenomenon as an integrated signal of independent single neurons was supported by patch clamp experiments as the number of action potentials (APs) per stimulus was decreased by addition of CO2. Addition of ISO on the other hand had no significant effect.Our results provide an explanation on the cellular level for anesthesia-dependent observations in previous studies of task-induced BOLD and resting state connectivity. They further inform selection of the adequate anesthetic regimen for a given combination of modalities.

Effects of different concentrations of isoflurane pretreatment on respiratory mechanics, oxygenation and hemodynamics in LPS-induced acute respiratory distress syndrome model of juvenile piglets

ABSTRACTPurpose: This study was prospectively designed to investigate the effects of different concentrations of isoflurane (ISO) pretreatment on respiratory mechanics, oxygenation, and hemodynamics in lipopolysaccharide (LPS)-induced acute respiratory distress syndrome (ARDS) model of juvenile piglets.Methods: Twenty-four piglets (9–14 kg, 5–6 weeks old) were randomly assigned to four groups (n = 6): LPS group, which was injected with LPS (20 μg/kg) to induce ARDS; 0.5 ISO-LPS, 1.0 ISO-LPS, and 1.3 ISO-LPS groups, which were pretreated with 0.5, 1.0, and 1.3 minimum alveolar concentrations (MAC) ISO for 30 min before immediate LPS infusion, respectively. After establishment of ARDS, respiratory mechanism, oxygenation and hemodynamics parameters were measured at baseline, and 0, 1, 2, 3, and 4 hours after induction of ARDS.Results: After induction of ARDS, there were increases in alveolar-arterial oxygen difference (A-aDO2), oxygenation index (OI), mean airway pressure (MAP), dead space-to-tidal volume ratio, heart rate (HR), dP/dtmax, extravascular lung water index, pulmonary vascular permeability index, and PaCO2, and decreases in PaO2/FIO2, respiratory rate (RR), dynamic lung compliance (Cdyn), mean arterial blood pressure (MABP) and systemic vascular resistance (SVR) compared with baseline (Ptime < 0.05). Pretreatment with 1.0 and 1.3 MAC ISO alleviated changes in dP/dtmax and PaCO2 at ARDS 0–2 hours, SVR at 0–3 hours, PaO2/FIO2, RR, and MABP at 1–2 hours, HR at 2–3 hours, A-aDO2 at 3–4 hours, and OI at 4 hours (Pgroup < 0.05). Conclusion: Pretreatment with 1.0 and 1.3 MAC ISO had protective effects on respiratory mechanics, oxygenation, and hemodynamics in piglets with LPS-induced ARDS.

2-deoxy-D-glucose enhances anesthetic effects in mice.

The mechanisms of general anesthesia by volatile drugs remain largely unknown. Mitochondrial dysfunction and reduction in energy levels have been suggested to be associated with general anesthesia status. 2-Deoxy-D-glucose (2-DG), an analog of glucose, inhibits hexokinase and reduces cellular levels of adenosine triphosphate (ATP). 3-Nitropropionic acid is another compound which can deplete ATP levels. In contrast, idebenone and L-carnitine could rescue deficits of energy. We therefore sought to determine whether 2-DG and/or 3-nitropropionic acid can enhance the anesthetic effects of isoflurane, and whether idebenone and L-carnitine can reverse the actions of 2-DG. C57BL/6J mice (8 months old) received different concentrations of isoflurane with and without the treatments of 2-DG, 3-nitropropionic acid, idebenone, and L-carnitine. Isoflurane-induced loss of righting reflex (LORR) was determined in the mice. ATP levels in H4 human neuroglioma cells were assessed after these treatments. Finally, 31P-magnetic resonance spectroscopy was used to determine the effects of isoflurane on brain ATP levels in the mice. 2-DG enhanced isoflurane-induced LORR (P = 0.002, N = 15). 3-Nitropropionic acid also enhanced the anesthetic effects of isoflurane (P = 0.005, N = 15). Idebenone (idebenone + saline versus idebenone + 2-DG: P = 0.165, N = 15), but not L-carnitine (L-carnitine + saline versus L-carnitine + 2-DG: P < 0.0001, N = 15), inhibited the effects of 2-DG on enhancing isoflurane-induced LORR in the mice, as evidenced by 2-DG not enhancing isoflurane-induced LORR in the mice pretreated with idebenone. Idebenone (idebenone + saline versus idebenone + 2-DG: P = 0.177, N = 6), but not L-carnitine (L-carnitine + saline versus L-carnitine + 2-DG: P = 0.029, N = 6), also mitigated the effects of 2-DG on reducing ATP levels in cells, as evidenced by 2-DG not decreasing ATP levels in the cells pretreated with idebenone. Finally, isoflurane decreased ATP levels in both cultured cells and mouse brains (β-ATP: P = 0.003, N = 10; β-ATP/phosphocreatine: P = 0.006, N = 10; β-ATP/inorganic phosphate: P = 0.001, N = 10). These results from our pilot studies have established a system and generated a hypothesis that 2-DG enhances anesthetic effects via reducing energy levels. These findings should promote further studies to investigate anesthesia mechanisms.

Brain-state dependent uncoupling of BOLD and local field potentials in laminar olfactory bulb

The neural activities of the olfactory bulb (OB) can be modulated significantly by internal brain states. While blood oxygenation level dependent functional MRI (BOLD-fMRI) has been extensively applied to study OB in small animals, the relationship between BOLD signals and electrophysiological signals remains to be elucidated. Our recent study has revealed a complex relationship between BOLD and local field potentials (LFP) signals in different OB layers during odor stimulation. However, no study has been performed to compare these two types of signals under global brain states. Here, the changes of BOLD and LFP signals in the glomerular, mitral cell, and granular cell layers of the OB under different brain states, which were induced by different concentrations of isoflurane, were sequentially acquired using electrode array and high-resolution MRI. It was found that under deeper anesthesia, the LFP powers in all layers were decreased but the BOLD signals were unexpectedly increased. Furthermore, the decreases of LFP powers were layer-independent, but the increases of BOLD signal were layer-specific, with the order of glomerular>mitral cell>granular cell layer. The results provide new evidence that the direct neural activity levels might not be correlated well with BOLD signals in some cases, and remind us that cautions should be taken to use BOLD signals as the index of neural activities.

Echocardiographic assessment of β-adrenoceptor stimulation-induced heart failure with reduced heart rate in mice

1. Chronic injection with the β-adrenoceptor (β-AR) agonist isoproterenol (ISO) has been commonly used as an animal model of β-AR-induced cardiac remodelling and heart failure. This ISO-treated model usually exhibits significantly decreased conscious heart rate (HR). However, the HR in treatment groups is usually adjusted to the same levels by anaesthesia to assess cardiac geometry and function. In the present study, we report a method of echocardiographic assessment that represents the true cardiac geometry and function under conditions of ISO withdrawal. 2. Briefly, C57BL/6 mice were treated with 5 mg/kg per day ISO for 12 weeks. Cardiac geometry and function were assessed by high-resolution echocardiography in vehicle (saline) - and ISO-treated mice that were either conscious or anaesthetized using different concentrations of isoflurane. 3. The cardiac β-AR response was decreased in ISO-treated mice, as evidenced by markedly decreased conscious HR. Vehicle- and ISO-treated mice did not differ in terms of cardiac geometry or function when HR was adjusted to the same level (400 b.p.m.) in both treatment groups, but cardiac geometry and function did differ when a low (1%) rather than high (1.5% or 2%) isoflurane concentration was used to adjust HR. Furthermore, 3 day ISO withdrawal eliminated the difference in conscious HR between the two groups. In addition, the groups differed in cardiac geometry and function regardless of the isoflurane concentration used. 4. In conclusion, using isoflurane to decrease the HR of treated groups to the same level may mask left ventricular dysfunction in ISO-treated mice. Withdrawal of ISO eliminated the difference in basal HR between the ISO-treated and control groups on echocardiography, allowing a more accurate assessment of cardiac pathological and functional changes.

Myocardial glucose uptake and metabolic rate measurements in mice with microPET imaging

Objective To establish a practical microPET imaging procedure for the measurement of myocardial metabolic rate of glucose (MRGlu) in mice.Methods Twenty wild-type BKS mice were divided into 4 groups by random number table method.The mice were anesthetized with different concentrations of isoflurane (1.3％,1.5％,1.8％,2.0％) at the temperature between 30 ℃ to 34 ℃.The respiratory rate and the physiologic condition were monitored for adjusting the most appropriate isoflurane concentration.Then,different volumes of saline were injected to the anesthetized mice and blood glucose concentrations were measured to test the optimal injection volume.Under the optimal operating condition,18 F-FDG were injected in a group of six mice and followed by microPET imaging.Left ventricular TAC was obtained by drawing ROI and myocardial glucose SUV was also calculated.Meanwhile,the TAC from venous sampling at different time points after 18F-FDG injection was generated.Kinetic Imaging System was used to estimate the coefficients and calculate the MRGlu(Ki ×Glu/LC; Ki =k1 ×k3/(k2+k3).One-way analysis of variance and q test were used to analyze the data.Results No movement was observed in non-fasted mice anesthetized with (1.5-1.8)％ isoflurane,and their respiratory rates were all over 80 per minute.The plasma glucose concentration showed no difference at each time point between the experimental group injected with 75 μl saline and the control group (F=1.215,P＞0.05).The plasma glucose concentration of mice injected with 150 μl saline exhibited statistically significant difference at 30 min (q =2.485,P=0.024),45 min (q =2.287,P=0.036) and 60 min (q =2.709,P =0.015).When the injection volume reached 300 μl,the blood glucose concentration increased remarkably at 45 min (q =2.435,P=0.027).Mice were maintained in good condition after injected with 18F-FDG ranging from 7.4 to 11.1 MBq within 75 μl volume,meanwhile clear and stable myocardial microPET images could also be obtained.The median myocardial SUV value was 11.88(9.71-14.93),Ki value was 0.19 (0.10-0.54) ml · min-1 · g-1and MRGlu value was 19.64 (5.55-23.28) mg · kg-1 · min-1at 45-55 min after 18F-FDG injection.Conclusion The microPET imaging may be a reliable,practical method to evaluate myocardial glucose uptake rate and metabolic rate in mice under the precondition of optimal isoflurane anesthetization (1.5％-1.8％)and small volume of 18F-FDG injection (≤＜75 μl). Key words: Myocardium; Deoxyglucose ; Metabolism ; Tomography, emission-computed ; Mice

Halogenated volatile anesthetics alter brain metabolism as revealed by proton magnetic resonance spectroscopy of mice in vivo

Halogenated volatile anesthetics (HVA) are widely used in medicine and research but their effects on brain metabolism in intact organisms are still largely unknown. Here, localized proton magnetic resonance spectroscopy (MRS) of anesthetized mice was applied to evaluate HVA effects on cerebral metabolites in vivo. Experimental protocols combined different concentrations of isoflurane, halothane, sevoflurane, and desflurane with known modulators of adrenergic, GABAergic, and glutamatergic neurotransmission. As a most striking finding, brain lactate increased in individual mice from 1.0 ± 0.6 mM (awake state) to 6.2 ± 1.5 mM (1.75% isoflurane). In addition, relative to total creatine, there were significant isoflurane-induced increases of alanine by 111%, GABA by 20%, choline-containing compounds by 20%, and myo-inositol by 10% which were accompanied by significant decreases of glucose by 51% and phosphocreatine by 9%. The elevation of lactate was most pronounced in the striatum. The HVA effects correlated with the respective minimal alveolar concentrations and were mostly reversible within minutes. The observed alterations are best explained by an HVA-induced stimulation of adrenergic pathways in conjunction with an inhibition of the respiratory chain. Apart from casting new light on cerebral energy metabolism, the present results challenge brain studies of HVA-anesthetized animals.

Effects of Isoflurane Concentration on Basic Echocardiographic Parameters of the Left Ventricle in Rats

Transthoracic echocardiography (TTE) has become an important modality for the assessment of cardiac structure and function in animal experiments. The acquisition of echocardiographic images in rats requires sedation/anesthesia to keep the rats immobile. Commonly used anesthetic regimens include intraperitoneal or inhalational application of various anesthetics. Several studies have compared the effects of anesthetic agents on echocardiographic parameters in rats; however, none of them examined the effects of different concentrations of inhalational anesthetics on echocardiographic parameters. Accordingly, the aim of this study was to examine the effects of different concentrations of isoflurane used for anesthesia during TTE examination in rats on basic echocardiographic parameters of left ventricular (LV) anatomy and systolic function. TTE examinations were performed in adult male Wistar rats (n=10) anesthetized with isoflurane at concentrations of 1.5-3 %. Standard echocardiograms were recorded for off-line analysis. An absence of changes in basic echocardiographic parameters of LV anatomy and systolic function was found under isoflurane anesthesia using concentrations between 1.5-2.5 %. An isoflurane concentration of 3 % caused a small, but statistically significant, increase in LV chamber dimensions without a concomitant change in heart rate or fractional shortening. For the purpose of TTE examination in the rat, our results suggest that isoflurane concentrations </= 2.5 % can be safely recommended.

Isoflurane enhances the expression of cytochrome C by facilitation of NMDA receptor in developing rat hippocampal neurons in vitro

This study examined the effects of clinically relevant concentrations of isoflurane on the amplitude of NMDA receptor current (I(NMDA)) and the expression of cytochrome C in cultured developing rat hippocampal neurons. The hippocampi were dissected from newborn Sprague-Dawley rats. Hippocampal neurons were primarily cultured for 5 days and then treated with different concentrations of isoflurane [(0.25, 0.5, 0.75, 1 minimum alveolar concentration (MAC))]. The peak of I(NMDA) was recorded by means of the whole cell patch clamp technique. The cytochrome C level was detected by Western blotting and quantitative real-time PCR. Our results showed that isoflurane (0.25, 0.5, 0.75 and 1 MAC) potentiated the amplitude of I(NMDA) by (116 ± 8.8)%, (122 ± 11.7)%, (135 ± 14.3)% and (132 ± 14.6)%, respectively, and isoflurane increased the mRNA expression of cytochrome C in a concentration-dependent manner. The cytochrome C mRNA expression reached a maximum after 0.5 MAC isoflurane stimulation for 6 h (P<0.05). It was concluded that isoflurane enhances the expression of cytochrome C in cultured rat hippocampal neurons, which may be mediated by facilitation of NMDA receptor.

The Potential Dual Effects of Anesthetic Isoflurane on Hypoxia-Induced Caspase-3 Activation and Increases in β-Site Amyloid Precursor Protein-Cleaving Enzyme Levels

β-Amyloid protein (Aβ) accumulation, caspase activation, apoptosis, and hypoxia-induced neurotoxicity have been suggested to be involved in Alzheimer disease neuropathogenesis. Aβ is produced from amyloid precursor protein through proteolytic processing by the aspartyl protease β-site amyloid precursor protein-cleaving enzyme (BACE) and γ-secretase. Inhaled anesthetics have long been considered to protect against neurotoxicity. However, recent studies have suggested that the inhaled anesthetic isoflurane may promote neurotoxicity by inducing caspase activation and apoptosis, and by increasing levels of BACE and Aβ. We therefore sought to determine whether isoflurane can induce concentration-dependent dual effects on hypoxia-induced caspase-3 activation and increases in BACE levels: protection versus promotion. H4 human neuroglioma cells were treated with hypoxia (3% O(2)) alone, different concentrations of isoflurane (0.5% and 2%), and the combination of hypoxia and 0.5% or 2% isoflurane. The levels of caspase-3 cleavage (activation), BACE, and Bcl-2 were determined by Western blot analysis. We show for the first time that treatment with 0.5% isoflurane for 8 hours attenuated, whereas treatment with 2% isoflurane for 8 hours enhanced, hypoxia-induced caspase-3 activation and increases in BACE levels. The 2% isoflurane treatment also enhanced a hypoxia-induced decrease in Bcl-2 levels. These results suggest a potential concept that isoflurane has dual effects (protection versus promotion) on hypoxia-induced toxicity, which may act through Bcl-2 family proteins. These findings could lead to more systematic studies to determine the potential dual effects of anesthetics on Alzheimer disease-associated neurotoxicity.

Lower Isoflurane Concentration Affects Spatial Learning and Neurodegeneration in Adult Mice Compared with Higher Concentrations

Volatile anesthetics such as isoflurane are widely used in clinical and research contexts. Concerns have been raised that the effects of these drugs on the central nervous system may result in long-term impairment after surgery or general anesthesia. Hence, this study aimed to detect how different isoflurane concentrations influence spatial learning and cell death in adult mice. Fifty-two C57BL/6 mice were randomly divided in four groups. Mice in three groups were exposed to different concentrations of isoflurane (1, 1.5, and 2%) for 1 h; the control group was not exposed to anesthesia. Five mice per group were killed 3 h after anesthesia to perform histopathologic and immunohistochemical analyses (hematoxylin-eosin staining; caspase-3 activation). Eight mice per group were used for behavioral tests (open field, T-maze spontaneous alternation, and water maze) on subsequent days. There were no differences between groups in the T-maze spontaneous alternation test or in the open field (no confounding effects of stress or locomotion). The group anesthetized with 1% isoflurane performed worse in the water maze task on day 1 (550.4 ±162.78 cm) compared with the control group (400.1 ± 112.88 cm), 1.5% isoflurane (351.9 ± 150.67 cm), and 2% isoflurane (364.5 ± 113.70 cm; P ≤ 0.05) and on day 3 (305.0 ± 81.75 cm) compared with control group (175.13 ± 77.00 cm) and 2% isoflurane (204.11 ± 85.75 cm; P ≤ 0.038). In the pyramidal cell layer of the region cornu ammonis 1 of the hippocampus, 1% isoflurane showed a tendency to cause more neurodegeneration (apoptosis) (61.4 ± 26.40, profiles/mm) than the group with 2% of isoflurane (20.6 ± 17.77, profiles/mm; P = 0.051). Low isoflurane concentration (1%) caused spatial learning impairment and more neurodegeneration compared with higher isoflurane concentrations. Results for mice receiving the latter concentrations were similar to those of control mice.

Effects of inhalation of different concentrations of isoflurane on hippocampal cytochrome c expression in aged rats

Objective To investigate the effects of inhalation of different concentrations of isoflurane on the expression of cytochrome c （ Cyt c） in hippocampus in aged rats. Methods Sixty-three aged male SD rats （20 months） weighing 500-600 g were randomly divided into 3 groups （ n = 21 each） ： control group inhaling 30% 02 for 2 h （group C） and 2 isoflurane groups anesthetized with 0.75% and 1.5% isoflurane in 30% Oz for 2 h respectively （group I1 and I2 ）. Arterial blood samples were obtained from 5 rats at 30 min, 1 and 2 h of anesthesia for blood gas analysis. Eight animals were killed at 24 h after anesthesia in each group. Their hippocampi were immediately removed for determination of Cyt c expression by immuno-histochcmistry and Western blot analysis. Cognitive function was assessed by Morris water maze test the day before experiment and once a day for 6 consecutive days starting from the 1st postoperative day. Results The Cyt c expression in hippocampus was significantly increased in I1 and I2 groups in a concentration-dependent manner as compared with group C. The escape latency was significantly prolonged and the frequency of crossing the original platform and the duration of staying at the original platform quadrant were decreased in group I1 and I2 compared with group C. Conclusion Inhalation of isoflurane anesthesia can decrease cognitive function through up-regulating the Cyt c expression in hippocampus in aged rats. Key words: Isoflurance ; Cytochromes c ; Hippocampus

Effects of isoflurane on neuronal apoptosis In aged rats

Objective To investigate the effects of different concentrations of isoflurane on neuronal apoptosis in hippocampus and cerebral cortex in the aged rats. Methods Ninety female pathogen-free SD rats 22-24 months old weighing 497-593 g were randomly divided into 3 groups: group Ⅰ oxygen group inhaled air enriched with 40% O2,groupⅡ and Ⅲ were anesthetized with 1.2% and 1.8% isoflurane for 3 h respectively. Morris water maze was used to assess the cognitive function. The animals were killed at 24 and 72 h and on the 7th day after emergence from anesthesia and their brains were removed. Neuronal apoptosis in hippocampus and cerebral cortex was detected by TUNEL. Results The average escape latency was significantly longer on the 2nd and 3rd day after isoflurane anesthesia in isoflurane group than in oxygen group (P < 0.05 ). The neuronal apoptesis rate was significantly higher in the 2 isoflurane group than in oxygen group (P < 0.05 or 0.01 ). Conclusion Isoflurane anesthesia can result in transient cognitive dysfunction, and the mechanism may be involved in apoptosis in cerebral cortex in the aged rats. Key words: Isoflurane; Aged; Cognition disorders; Neurons; Apoptosis

This Month in Anesthesiology

The authors examined the epidemiology of perioperative AIS in three common surgeries in adult patients: hemicolectomy, total hip replacement, and lobectomy/segmental lung resection. Data were extracted from a national database in the United States for the years 2000 to 2004. Patients with perioperative AIS were identified as were comorbid conditions that may be risk factors for perioperative AIS. Those who developed perioperative AIS were as follows: 0.7% of hemicolectomy patients, 0.2% of total hip replacement patients, and 0.6% of lobectomy/segmental lung resection patients. For patients older than 65, AIS rose to 1.0% for hemicolectomy, 0.3% for hip replacement, and 0.8% for pulmonary resection. Perioperative AIS is an important source of morbidity and mortality associated with noncardiac, nonvascular surgery particularly in elderly patients. See the accompanying Editorial View on page 209 Anesthetic requirements of the fruit fly mutant minisleeper and other Drosophila mutants were determined based on the response of the flies at different concentrations of isoflurane and sevoflurane. The average amount of daily sleep in wild-type Drosophila was 965 minutes but only 584 minutes in minisleeper mutant flies. The EC50values in wild-type Drosophila were 0.706 for isoflurane and 1.298 for sevoflurane; however, EC50values in the minisleeper mutant flies for isoflurane and sevoflurane were 1.306 and 2.013, respectively. A single-gene mutation in Drosophila that causes an extreme reduction in daily sleep is responsible for a significant increase in the requirement of volatile anesthetics. This study has implications for sleep and mechanisms of anesthesia and sedation.Hyperglycemia in perioperative patients has been identified as a risk factor for morbidity and mortality. Intensive insulin therapy (IIT) has been shown to reduce morbidity and mortality among the critically ill, decrease infection rates and improve survival after cardiac surgery, and improve outcomes in acute neurological injury and acute myocardial infarction. However, severe hypoglycemia and adverse events associated with IIT are a concern. The authors summarize the mechanisms and rationale of hyperglycemia and IIT, and review the evidence behind the use of IIT in the perioperative period. While avoidance of hyperglycemia is clearly beneficial, the appropriate glucose target and specific subpopulations that might benefit from IIT have yet to be identified. See the accompanying Editorial View on page 204 The authors hypothesized that the use of factor XIII would delay the decrease in clot firmness in high-risk patients with high fibrin monomer levels. This concept was tested in a prospective, randomized, double-blind, placebo-controlled trial in elective cancer surgery. Patients were randomized to receive factor XIII (30 U/kg) or placebo and standard therapy. During the interim analysis, clot firmness was evaluated. Patients receiving placebo lost 38% firmness whereas those receiving factor XIII did not. This proof of concept study confirmed that patients at high risk for intraoperative blood loss showed reduced loss of clot firmness when factor XIII was administered. Further clinical trials are needed to assess this treatment on blood loss and use of blood products. See the accompanying Editorial View on page 212 An updated report by the American Society of Anesthesiologists Task Force on Neuraxial Opioids.In multivariate models, body mass index is a weak but significant predictor of difficult and failed intubation.Tighter glucose control may improve renal function in potential organ donors. See the accompanying Editorial View on page 207 Early transfusion is an independent predictor of adult respiratory distress syndrome in adult trauma. See the accompanying Editorial View on page 216

Comparison of the effects of isoflurane and sevoflurane on protein tyrosine phosphorylation‐mediated vascular contraction

Isoflurane induces greater effects on vasodilation and decreasing blood pressure than sevoflurane. Tyrosine kinase-catalyzed protein tyrosine phosphorylation plays an important role in regulating vascular smooth muscle contraction. The aim of the present study was to compare the effects of isoflurane and sevoflurane on tyrosine phosphorylation-mediated vascular constriction, by assessing the degree of sodium orthovanadate (Na(3)VO(4), tyrosine phosphatase inhibitor)-induced contraction and protein tyrosine phosphorylation of rat aortic smooth muscle. Na(3)VO(4)-induced contraction and protein tyrosine phosphorylation of rat aortic smooth muscle were measured in the presence of genistein, a tyrosine kinase inhibitor, and different concentrations of isoflurane and sevoflurane, using isometric force measurement and Western blot, respectively. Na(3)VO(4) (10(-4) M) induced sustained contraction and tyrosine phosphorylation of substrates that were both markedly attenuated in the presence of genistein (5 x 10(-5) M). Isoflurane and sevoflurane dose-dependently (1, 2, 3 MAC) attenuated the Na(3)VO(4)-induced contraction (P < 0.05-0.005, n = 8), with a greater degree of inhibition by isoflurane than sevoflurane at 2 MAC (P < 0.01) and 3 MAC (P < 0.05). Both anesthetics also attenuated the total band density of the Na(3)VO(4)-induced, tyrosine-phosphorylated substrates in a concentration-dependent manner (P < 0.05-0.005, n = 4), with much greater attenuation by isoflurane than sevoflurane at 1 and 2 MAC (P < 0.05), respectively. The results of the present study demonstrate that isoflurane exhibits a greater degree of inhibition on the Na(3)VO(4)-stimulated contraction and protein tyrosine phosphorylation of rat aortic smooth muscle compared with sevoflurane. These findings suggest that isoflurane depresses the protein tyrosine phosphorylation-mediated contraction of vascular smooth muscle to a greater degree than sevoflurane.