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Reversal of morphine-induced catalepsy by naloxone microinjections into brain regions with high opiate receptor binding: a preliminary report.

The relationship between opiate binding density and morphine-induced catalepsy was estimated via dose-response analysis of the brain sites in which naloxone microinjections reversed the catalepsy induced by intraperitoneal morphine. One-hundred forty-one experimentally naive male Long-Evans rats were implanted with chemical microinjection guide cannulae aimed for various high-to-moderate binding density areas within caudate nucleus, central gray matter, thalamus, hypothalamus, amygdala, and frontal cortex as well as low density sites in pyriform cortex and various fiber tracts. Overall, 48 out of 91 animals microinjected with naloxone in brain sites having high-to-moderate density of opiate binding showed reversal of the cataleptic response. Dose-response effects were found in all 6 high-to-moderate density sites: ranging from 85% reversals at 100 mcg naloxone over all sites to 34% reversals at 0.01 mcg naloxone. There were no reversals out of 38 naloxone microinjection in brain sites having a low density of opiate binding and no reversals out of 18 saline microinjections in either high-to-moderate or low opiate binding density loci. These results suggest a role for limbic and basal ganglia portions of the opiate system in a motor aspect of narcotic action. We speculate that these loci may also play a role in the motor expression of the response to the analgesic and euphoric actions of morphine to supplement actions mediated through periventricular structures.

Animals↗

Amphetamine and the overtraining reversal effect.

Rats were trained in a Y-maze on a two choice simultaneous brightness discrimination with light as S+ and dark as S- (position irrelevant). Animals in the Mastery group were trained until they reached criterion and were then switched to reversal, where the reinforcement contingencies of the original training were reversed. Animals in the Overtraining group received a further 110 trials before being switched to reversal. The administration of 1 mg/kg d-amphetamine facilitated dramatically reversal learning in Mastery group. Overtraining improved reversal in saline injected animals and slowed down reversal in amphetamine-treated animals. The drug also facilitated the acquisition of the initial brightness discrimination.

Amphetamine↗

Extraction of lysozyme, alpha-chymotrypsin, and pepsin into reverse micelles formed using an anionic surfactant, isooctane, and water.

The extraction of lysozyme, alpha-chymotrypsin, and pepsin from buffered salt solutions into reverse micelles was examined at different pH values and surfactant concentrations. The reverse micelles was formed by mixing aqueous buffer supplemented with KCl and an organic phase of isooctane(2,2,4-trimethylpentane), containing the anionic surfactant, Aerosol O. T. (dioctyl ester of sodium sulfosuccinic acid). The technique of dynamic laser scattering was used to measure the size of reverse micelles which were in equilibrium with the aqueous phase. It was found that the size of the reverse micelles decreased with increasing ionic strength but increased with increasing AOT concentration. In the process of extraction, the reverse micelles might have rearranged themselves to host the protein. The sizes of protein-filled and -unfilled reverse micelles were different, and an open equilibrium could be reached between them. Under the extraction conditions, only a small number of micelles were found to contain protein.

Chymotrypsin↗

A new strategy for the study of oligomeric enzymes: gamma-glutamyltransferase in reversed micelles of surfactants in organic solvents.

A heterodimeric enzyme (gamma-glutamyltransferase) was studied in the reversed micellar medium of Aerosol OT (AOT) in octane. As was shown earlier, the size (radius) of inner cavity of the AOT-reversed micelles is determined by their hydration degree, i.e., [H2O]/[AOT] molar ratio, in the system. Owing to this, the dependence of hydrolytic, transpeptidation and autotranspeptidation activities of the enzyme on the hydration degree was investigated using L- and D-isomers of gamma-glutamyl(3-carboxy-4-nitro)anilide and glycylglycine as substrates. For all of the reaction types, the observed dependences are curves with three optima. The optima are found at the hydration degrees, [H2O]/[AOT] = 11, 17 and 26 when the inner cavity radii of reversed micelles are equal to the size of light (Mr 21,000) and heavy (Mr 54,000) subunits of gamma-glutamyltransferase, and to their dimer (Mr 75,000), respectively. Ultracentrifugation experiments showed that a change of the hydration degree resulted in a reversible dissociation of the enzyme to light and heavy subunits. The separation of light and heavy subunits of gamma-glutamyltransferase formed in reversed micelles was carried out and their catalytic properties were studied. The two subunits catalyze hydrolysis and transpeptidation reactions; autotranspeptidation reaction is detected only in the case of the heavy subunit. These findings imply that the reversed micelles of surfactants in organic solvents function as the matrices with adjustable size permitting to regulate the supramolecular structure and the catalytic activity of oligomeric enzymes.

Catalysis↗

Fluorescence dynamics of staphylococcal nuclease in aqueous solution and reversed micelles.

The dynamical fluorescence properties of the sole tryptophan residue (Trp-140) in Staphylococcus aureus nuclease (EC 3.1.31.1) have been investigated in aqueous solution and reversed micelles composed of either sodium bis(2-ethylhexyl)sulfosuccinate (AOT) in isooctane or cetyltrimethylammonium chloride (CTAC) in isooctane/hexanol (12:1 by volume). The fluorescence decay of nuclease in the different environments can be described by a trimodal distribution of fluorescence lifetimes at approx. 0.5, 1.5 and 5.0 ns. The relative amplitudes depend on the environment. For pH 9.0 solutions the contribution of the two shortest lifetime components in the distribution is largest for AOT and smallest for CTAC reversed micelles. There is reasonable agreement between the average fluorescence lifetime and the fluorescence quantum efficiency confirming a significant fluorescence quenching in AOT reversed micelles. Fluorescence anisotropy decay revealed that the tryptophan environment in aqueous nuclease solutions is rigid on a nanosecond timescale. When nuclease was entrapped into reversed micelles the tryptophan gained some internal flexibility as judged from the distinct presence of a shorter correlation time. The longer correlation time reflected the rotational properties of the protein-micellar system. Modulation of the overall charge of nuclease (isoelectric point pH 9.6) by using buffer of pH 9.0 and pH 10.4, respectively, and of the size of empty micelles by selecting two values of the water to surfactant molar ratio, had only a minor effect on the rotational properties of nuclease in the positively charged reversed micelles. Encapsulation of nuclease in anionic reversed micelles resulted in the development of protein bound to aggregated structures which are immobilised on a nanosecond timescale. According to far UV circular dichroism results the secondary structure of nuclease only followed the already published pH-dependent changes. Encapsulation had no major effect on the overall secondary structure.

Cetrimonium↗

Vasopressin-sensitive adenylate cyclase. Reversibility of hormonal activation.

The reversibility of adenylate cyclase activation induced by vasopressin was studied by reducing the concentration of active peptide in contact with kidney medullo-papillary membranes. Reversibility of hormonal activation was only partial. The use of antagonists failed to demonstrate the reversibility of an adenylate cyclase activation induced by high affinity agonists. When antagonist was added after the agonist to membranes, a non-competitive inhibition was apparent. Active peptide was also eliminated from the incubation medium by treatment with agents capable of reducing the disulfide bridge of the hormonal molecule. Direct effects of reducers on adenylate cyclase activity were measured on enzyme activation induced by peptides lacking a disulfide bridge. There was no apparent correlation between the abilities of different reducers to inactivate free peptide in solution and their abilities to promote the reversibility of hormone-induced enzyme activation. Upon the addition of dithiothreitol, enzyme activity could be lowered to basal value and adenylate cyclase was again fully stimulatable. However, when dithiothreitol addition to stiumlated enzyme was combined with a 60-fold dilution of the incubation medium, no reversibility of hormonal activation occurred. These results illustrate that the processes involved in adenylate cyclase activation are only partially reversible.

Adenylyl Cyclases↗

Reversible depression of the reticuloendothelial system by liposomes.

The effect of various doses of different types (reverse phase evaporation vesicles and small unilamellar vesicles) of intravenously injected liposomes on reticuloendothelial activity, as measured by the blood clearance rate of intravenously injected carbon, was investigated. Also the effect of pretreatment with reverse phase evaporation vesicles on blood clearance and tissue distribution of a second dose of similar vesicles was determined. For all concentrations used reverse phase evaporation vesicles caused a reduction in reticuloendothelial activity at least up to 4 h after injection. 24 h after administration the rate of carbon clearance returned to the control level. On the contrary small unilamellar vesicles did not block reticuloendothelial activity. Pretreatment with reverse phase evaporation vesicles (250 mumol/kg) caused an increased blood level and a decreased hepatic uptake of a second dose of the vesicles, injected 1 h after the first dose. This seems to be due to a depression of reticuloendothelial activity and not to a depletion of opsonins. Pretreatment with small unilamellar vesicles (250 mumol/kg) had no significant influence on the tissue distribution of a second dose of vesicles. Our results clearly indicate that reverse phase evaporation vesicles cause a reversible depression of reticuloendothelial activity and this depression seems to be induced by a saturation of reticuloendothelial cells with liposomes.

Animals↗

Effects of noradrenergic DSP4 lesion on the effectiveness of pilocarpine in reversing scopolamine-induced amnesia.

We studied the effectiveness of pilocarpine in reversing the scopolamine-induced water maze learning deficit (increase in escape latencies, decrease in spatial bias) in control and DSP4- (a noradrenergic neurotoxin) lesioned rats. The water maze acquisition deficit (escape latency, first spatial bias) induced by scopolamine 0.8 mg/kg was augmented by DSP4 treatment. The water maze performance deficit induced by scopolamine was reversed by pilocarpine 4 mg/kg in both DSP4-lesioned and control rats. A smaller dose of pilocarpine (1 mg/kg) did not reverse scopolamine-induced acquisition deficit in either control or DSP4-lesioned rats. Analysis of the second spatial bias test measured 2 weeks after training revealed that pilocarpine 4 mg/kg reversed scopolamine-induced retention deficit in control and DSP4-lesioned rats. Pilocarpine 1 mg/kg reversed scopolamine-induced retention performance deficit during the second spatial bias test in control but not in DSP4-lesioned rats. The present results suggest that 1) noradrenergic and cholinergic systems may interact in the regulation of spatial acquisition and retention and 2) the effectiveness of cholinergic drugs in reversing scopolamine-induced spatial retention deficit may be affected by noradrenergic lesioning.

Amnesia↗

Reversal of diastolic cerebral blood flow in infants without brain death.

The reversal of diastolic cerebral blood flow has been regarded as a characteristic waveform of brain death and a useful confirming sign. We report 2 patients who had diastolic flow reversal but survived. One, a 1-month-old boy with status epilepticus, had reversal of diastolic cerebral blood flow detected by Doppler ultrasound soon after admission. Reversal disappeared after medical management for increased intracranial pressure and seizure control. He recovered without sequelae. The other, a 6-month-old girl with choroid plexus papilloma, had reversal of diastolic flow during abrupt clinical deterioration. Emergent surgical removal of the tumor was performed and she survived with hemiparesis and psychomotor retardation. Our patients demonstrated that even in the presence of diastolic reversal of cerebral blood flow, prompt and effective treatment can avoid a fatal outcome.

Blood Flow Velocity↗

Accelerated decline in lung function and impaired reversibility with salbutamol in asthmatic patients with chronic hepatitis C virus infection: a 6-year follow-up study.

PURPOSE: Chronic hepatitis C virus (HCV) infection may have adverse effects on pulmonary function in patients with chronic obstructive pulmonary disease. This prospective study was designed to determine whether chronic HCV infection affects decline in lung function and airway responses to salbutamol in asthmatic patients. METHODS: Interferon alpha (6 MIU three times a week for 6 months) was given to 55 HCV-positive asthmatic patients, 18 of whom had a virologic response to interferon. Pre- and postbronchodilator forced expiratory volume in 1 second (FEV(1)) and reversibility with salbutamol or oxitropium at years 1, 3, and 6 after interferon therapy were examined. RESULTS: We found a significant decrease in pre- and postbronchodilator FEV(1) from year 1 to years 3 and 6 only in interferon nonresponders. Reversibility with salbutamol at years 3 and 6 was significantly lower in interferon nonresponders than in HCV-negative patients (P <0.0001) and interferon responders (P <0.0001). Moreover, there was a steep decline in reversibility with salbutamol during the follow-up period only in interferon nonresponders. In contrast, reversibility with oxitropium at years 3 and 6 was significantly higher in interferon nonresponders than in HCV-negative patients and interferon responders, and there was a steep increase in reversibility with oxitropium only in interferon nonresponders. In addition, declines in the diffusing capacity of the lung for carbon monoxide during follow-up were significantly greater in interferon nonresponders than in HCV-negative patients and interferon responders. CONCLUSION: Chronic HCV infection is associated with an accelerated decline in lung function and impaired reversibility with salbutamol among asthmatic patients who do not respond to interferon therapy.

Albuterol↗

PI3 kinase is involved in cocaine behavioral sensitization and its reversal with brain area specificity.

Phosphatidylinositol 3-kinase (PI3K) is an important signaling molecule involved in cell differentiation, proliferation, survival, and phagocytosis, and may participate in various brain functions. To determine whether it is also involved in cocaine sensitization, we measured the p85alpha/p110 PI3K activity in the nuclear accumbens (NAc) shell, NAc core, and prefrontal cortex (PFC) following establishment of cocaine sensitization and its subsequent reversal. Naïve rats were rank-ordered and split into either daily cocaine or saline pretreatment group based on their locomotor responses to an acute cocaine injection (7.5 mg/kg, i.p.). These two groups were then injected with cocaine (40 mg/kg, s.c.) or saline for 4 consecutive days followed by 9-day withdrawal. Cocaine sensitization was subsequently reversed by 5 daily injections of the D1/D2 agonist pergolide (0.1 mg/kg, s.c.) in combination with the 5-HT3 antagonist ondansetron (0.2 mg/kg, s.c., 3.5h after pergolide injection). After another 9-day withdrawal, behavioral cocaine sensitization and its reversal were confirmed with an acute cocaine challenge (7.5 mg/kg, i.p.), and animals were sacrificed the next day for measurement of p85alpha/p110 PI3K activity. Cocaine-sensitized animals exhibited increased PI3K activity in the NAc shell, and this increase was reversed by combined pergolide/ondansetron treatment, which also reversed behavioral sensitization. In the NAc core and PFC, cocaine sensitization decreased and increased the PI3K activity, respectively. These changes, in contrast to that in the NAc shell, were not normalized following the reversal of cocaine-sensitization. Interestingly, daily injections of pergolide alone in saline-pretreated animals induced PI3K changes that were similar to the cocaine sensitization-associated changes in the NAc core and PFC but not the NAc shell; furthermore, these changes in saline-pretreated animals were prevented by ondansetron given 3.5h after pergolide. The present study suggests that selective enhancement of the PI3K activity in the NAc shell may be one of key alterations underlying the long-term cocaine sensitization. To the extent cocaine sensitization is an important factor in human cocaine abuse, pharmacological interventions targeted toward the NAc shell PI3K alteration may be useful in cocaine abuse treatment.

Animals↗

Higher order structure of proteins solubilized in AOT reverse micelles.

The higher order structure of proteins solubilized in an bis(2-ethylhexyl) sulfosuccinate sodium (AOT) reverse micellar system was investigated. From circular dichroic (CD) measurement, CD spectra of cytochrome c, which is solubilized at the interface of reverse micelles, markedly changed on going from buffer solution to the reverse micellar solution, and the ellipticity values in the far- and near-UV regions decreased with decreasing the water content (W0: molar ratio of water to AOT), indicating that the secondary and tertiary structures of cytochrome c changed with the water content. The ellipticity of ribonuclease A, which is solubilized in the center of micellar water pool, in the near-UV region was dependent on W0 and became minimum when W0 of ca. 8 while the ellipticity in the far-UV region was almost constant, indicating that the tertiary structure of ribonuclease A was affected by the water content, but the secondary structure was conserved. The degree of curvature of the micellar interface appears to influence the protein structure because the reverse micelle size is linearly proportional to the W0 value. As evidence of this, when the micelle size was comparable to the protein's dimensions, the structures were more affected by the water content. Judging from the dependence of the factor influencing the protein structure on the protein species, the location of solubilized protein in reverse micelles is significantly related to whether the protein structure in the system is affected by the micellar interface. In the cases of cytochrome c and lysozyme, the ellipticity against W0 was dependent on the AOT concentration. In contrast, ribonuclease A gave very similar ellipticity values whatever the AOT concentration. In the n-hexane micellar system, cytochrome c exhibited lower ellipticity values and ribonuclease A in the lower W0 range (W0<ca. 8) higher ellipticity values. These results indicated that the interaction between the protein and the micellar interface is a dominant factor influencing the protein structure in reverse micelles, and that it is governed by the location of solubilized proteins and the state of the micellar interface.

Circular Dichroism↗

Necker cube reversals during long-term EEG recordings: sub-bands of alpha activity.

Reversible figures, such as the Necker cube, make up a well-known class of visual phenomena in which an invariant stimulus pattern gives rise to at least two different perceptual interpretations. A better understanding of the neurophysiological processes underlying perceptual reversals might help to disentangle bottom-up from top-down influences on multistable perception. Recently, we reported alpha activity decrease during multistable visual perception. The aim of the present study was to define more specifically the functional roles of the EEG alpha band during the perception of Necker cube reversals by subdividing the extended alpha band into three sub-bands (lower-1 alpha, lower-2 alpha, upper alpha). We employed a long-term recording condition, during which 10 healthy participants observed the Necker cube for approximately 60 min and responded by pressing a button to any perceived reversal. The results showed a reversal induced alpha desynchronization for the lower alpha bands, with the lower-2 alpha desynchronization differing across the time course of the experiment. The upper alpha band demonstrated no reliable effects. It is concluded that the lower-1 alpha desynchronization reflects an automatic arousal reaction which triggers attentional processing in a bottom-up manner, whereas the lower-2 alpha desynchronization is related to attentional processes that are achieved by top-down control with limited resources. The lack of reliable effects in the upper alpha band is presumably due to the relatively low semantic task demands in figure reversal.

Adult↗

A new electrocardiographic algorithm to differentiate upper loop re-entry from reverse typical atrial flutter.

OBJECTIVES: This study was performed to differentiate upper loop re-entry (ULR) from reverse typical atrial flutter (AFL). BACKGROUND: Right atrial ULR and reverse typical AFL have different mechanisms and ablation strategies, but similar electrocardiographic characteristics. METHODS: This study included 26 patients with reverse typical AFL and 20 patients with ULR. The noncontact mapping system (EnSite-3000, Endocardial Solutions, St. Paul, Minnesota) was used to confirm diagnosis and guide successful radiofrequency ablation. Flutter wave polarity and amplitude in the 12-lead surface electrocardiogram were determined by two independent electrophysiologists. RESULTS: The flutter wave polarity in leads I and aVL was significantly different between the reverse typical AFL and ULR groups (p < or = 0.001). Voltage measurement revealed significant differences between reverse typical AFL and ULR in leads I, II, aVR, aVF, V1, and V2 (p < 0.001). A new diagnostic algorithm based on negative or isoelectric/flat flutter wave polarity and amplitude < or =0.07 mV in lead I was useful for diagnosis of ULR, with an accuracy of 90% to 97%, a sensitivity of 82% to 100%, and a specificity of 95%. CONCLUSIONS: Polarity and voltage measurement of flutter wave in lead I can differentiate reverse typical AFL from ULR.

Adult↗

Fractional flow reserve of infarct-related arteries identifies reversible defects on noninvasive myocardial perfusion imaging early after myocardial infarction.

OBJECTIVES: We hypothesized that fractional flow reserve (FFR) of an infarct-related artery (IRA) early after myocardial infarction (MI) identifies inducible ischemia on noninvasive imaging. BACKGROUND: Early after MI, IRAs frequently have angiographically indeterminant lesions. Whether FFR can detect reversible perfusion defects early after MI when dynamic microvascular abnormalities are present is not known. METHODS: Rest and dipyridamole (DP)-stress 99mTc sestamibi single-photon emission computed tomography (SPECT) were performed in 48 patients 3.7 +/- 1.3 days after MI, with 23 patients undergoing concurrent myocardial contrast echocardiography (MCE). Angiography, FFR, and percutaneous coronary intervention (PCI) of the IRA (as necessary) were subsequently performed. Follow-up SPECT was performed 11 weeks after PCI to identify true reversibility on baseline SPECT. RESULTS: The sensitivity, specificity, positive and negative predictive value, and concordance of FFR < or =0.75 for detecting reversibility on SPECT were 88%, 50%, 68%, 89%, and 71% (chi-square <0.001), respectively; which improved to 88%, 93%, 88%, 93%, and 91% (chi-square <0.001), respectively, for the detection of true reversibility. The corresponding values of FFR < or =0.75 for detecting reversibility on DP-MCE were 90%, 100%, 100%, 75%, and 93% (chi-square <0.001), respectively, and on either SPECT or MCE were 88%, 93%, 91%, 91%, and 91% (chi-square <0.001), respectively. The optimal FFR value for discriminating inducible ischemia on noninvasive imaging was 0.78. CONCLUSIONS: Fractional flow reserve of the IRA accurately identifies reversibility on noninvasive imaging early after MI. These findings support the utility of FFR early after MI.

Aged↗

Effect of hexanol on the reversed micelles of Span 85 modified with Cibacron Blue F-3GA for protein solubilization.

Sorbitan trioleate (Span 85) modified with Cibacron Blue F-3GA (CB) was used as an affinity surfactant (CB-Span 85) to form affinity-based reversed micelles in n-hexane. It was found that the addition of hexanol to the reversed micellar system resulted in a significant increase in water content and hydrodynamic radius of the affinity-based reversed micelles. Moreover, the reversed micelles with hexanol revealed broader aggregation number distribution and larger average aggregation number than the reversed micelles without hexanol addition. This is considered to be due to the decreases in the micellar curvature and rigidity of the micellar interfacial layer and the increase in the micellar interfacial fluidity. Consequently, the solubilization capacity of lysozyme increased about 70% in the reversed micellar solution with 3 vol% hexanol. On the other hand, the capacity of BSA was only 30% increased under the same conditions due to its larger molecular size than lysozyme. Kinetic analysis revealed that the increase in the micellar interfacial fluidity in the presence of hexanol resulted in faster release of lysozyme from the micelles, thus leading to an increase of the overall volumetric mass transfer coefficient in the back extraction.

Hexanols↗

Reversing cell polarity: evidence and hypothesis.

The long, rod-shaped cells of myxobacteria are polarized by their gliding engines. At the rear, A-engines push while pili pull the front end forward. An hypothesis is developed whereby both engines are partially dis-assembled, then re-assembled at the opposite pole when cells reverse their movement direction. Reversals are induced by an Mgl G-protein switch that controls engine polarity. The switch is driven by an oscillatory circuit of Frizzy proteins. In growing cells, the circuit gives rise to an occasional reversal that makes swarming possible. Then, as myxobacteria begin fruiting body development, a rising level of C-signal input drives the oscillator and changes the reversal pattern. Cells reverse regularly every eight minutes in traveling waves, the reversal period is then prolonged enabling cells to form streams that enlarge tiny random aggregates into fruiting bodies.

Bacterial Proteins↗

Gestural recovery and the role of forward and reversed syllabic repetitions as stuttering inhibitors in adults.

Instead of being the core stuttering 'problem', syllabic repetitions may be a biological mechanism, or 'solution', to the central involuntary stuttering block. Simply put, stuttering is an endogenous transitory state of 'shadowed speech', a choral speech derivative that allows for a neural release of the central block. To investigate this possibility, 14 adults who stutter read while listening to forward fluent speech, reversed fluent speech, forward stuttered speech, and reversed stuttered speech. All conditions induced significant degrees of stuttering inhibition when compared to a control condition. However, the reversed fluent condition was less powerful than the other three conditions ( approximately 42% vs. approximately 65%) for inhibiting stuttering. Stuttering inhibition appears to proceed by 'gestural recovery', made possible by the presence of an exogenous or 'second' set of speech gestures and engagement of mirror neurons. When reversed fluent speech was used, violations in normal gesture-time relationships (i.e., normal speech entropy) resulted in gestural configurations that apparently were inadequately recovered, and therefore, were not as conducive to high levels of stuttering inhibition. In contrast, high levels of encoding found in the simple syllabic structures of stuttered speech allowed its forward and reversed forms to be equally effective for gestural recovery and stuttering inhibition. The reversal of repeated syllables did not appear to significantly degrade the natural gesture-time relationships (i.e., they were perceptually recognizable). Thus, exogenous speech gestures that displayed near normal gestural relationships allowed for easy recovery and fluent productions via mirror systems, suggesting a more choral-like nature. The importance of syllabic repetitions is highlighted: both their perceived (exogenous) and produced (endogenous) forms appear to be fundamental, surface acoustic manifestations for central stuttering inhibition via the engagement of mirror neurons.

Adult↗