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Biomedical subjects

S Rouhani

Publications and source records attributed to S Rouhani.

18 recordsLinked to original sources
PubMed2001

Crystal structure of the D85S mutant of bacteriorhodopsin: model of an O-like photocycle intermediate.

Crystal structures are reported for the D85S and D85S/F219L mutants of the light-driven proton/hydroxyl-pump bacteriorhodopsin. These mutants crystallize in the orthorhombic C222(1) spacegroup, and provide the first demonstration that monoolein-based cubic lipid phase crystallization can support the growth of well-diffracting crystals in non-hexagonal spacegroups. Both structures exhibit similar and substantial differences relative to wild-type bacteriorhodopsin, suggesting that they represent inherent features resulting from neutralization of the Schiff base counterion Asp85. We argue that these structures provide a model for the last photocycle intermediate (O) of bacteriorhodopsin, in which Asp85 is protonated, the proton release group is deprotonated, and the retinal has reisomerized to all-trans. Unlike for the M and N photointermediates, where structural changes occur mainly on the cytoplasmic side, here the large-scale changes are confined to the extracellular side. As in the M intermediate, the side-chain of Arg82 is in a downward configuration, and in addition, a pi-cloud hydrogen bond forms between Trp189 NE1 and Trp138. On the cytoplasmic side, there is increased hydration near the surface, suggesting how Asp96 might communicate with the bulk during the rise of the O intermediate.

Amino Acid Substitution↗
PubMed2001

Protective effects of cardiotrophin-1 adenoviral gene transfer on neuromuscular degeneration in transgenic ALS mice.

Amyotrophic lateral sclerosis (ALS) is mainly a sporadic neurodegenerative disorder characterized by loss of cortical and spinal motoneurons. Some familial ALS cases (FALS) have been linked to dominant mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). Transgenic mice overexpressing a mutated form of human SOD1 with a Gly93Ala substitution develop progressive muscle wasting and paralysis as a result of spinal motoneuron loss and die at 5 to 6 months. We investigated the effects of neurotrophic factor gene delivery in this FALS model. Intramuscular injection of an adenoviral vector encoding cardiotrophin-1 (CT-1) in SOD1G93A newborn mice resulted in systemic delivery of CT-1, supplying motoneurons with a continuous source of trophic factor. CT-1 delayed the onset of motor impairment as assessed in the rotarod test. Axonal degeneration was slowed and skeletal muscle atrophy was largely reduced by CT-1 treatment. By monitoring the amplitude of the evoked motor response, we showed that the time-course of motor impairment was significantly decreased by CT-1 treatment. Thus, adenovirus-mediated gene transfer of neurotrophic factors might delay neurogenic muscular atrophy and progressive neuromuscular deficiency in ALS patients.

Adenoviridae↗
PubMed2001

Structure of an early intermediate in the M-state phase of the bacteriorhodopsin photocycle.

The structure of an early M-intermediate of the wild-type bacteriorhodopsin photocycle formed by actinic illumination at 230 K has been determined by x-ray crystallography to a resolution of 2.0 A. Three-dimensional crystals were trapped by illuminating with actinic light at 230 K, followed by quenching in liquid nitrogen. Amide I, amide II, and other infrared absorption bands, recorded from single bacteriorhodopsin crystals, confirm that the M-substate formed represents a structure that occurs early after deprotonation of the Schiff base. Rotation about the retinal C13-C14 double bond appears to be complete, but a relatively large torsion angle of 26 degrees is still seen for the C14-C15 bond. The intramolecular stress associated with the isomerization of retinal and the subsequent deprotonation of the Schiff base generates numerous small but experimentally measurable structural changes within the protein. Many of the residues that are displaced during the formation of the late M (M(N)) substate formed by three-dimensional crystals of the D96N mutant (Luecke et al., 1999b) are positioned, in early M, between their resting-state locations and the ones which they will adopt at the end of the M phase. The relatively small magnitude of atomic displacements observed in this intermediate, and the well-defined positions adopted by nearly all of the atoms in the structure, may make the formation of this structure favorable to model (simulate) by molecular dynamics.

Bacteriorhodopsins↗
PubMed2001

Multiple-point variance analysis for optimal adjustment of a monitoring network.

The selection of sampling sites is one of the major tasks in the design of a monitoring network. Many environmental networks suffer from either insufficient information or redundant information. This study presents a new, effective algorithm that addresses the issues of insufficient and reduction information. The new algorithm is denoted as Multiple-Point Variance Analysis (MPV). MPV includes both Multiple-Point Variance Reduction Analysis (MPVR) for adding information-effectives sites, and Multiple-Point Variance Increase Analysis (MPVI) for deleting information-redundant sites. The MPVR and MPVI equations are verified under two hypothetical cases. The optimal procedures of this new algorithm include determination of simultaneous additions or deletions of groups of sampling points. These proposed optimization procedures eliminate the need for any spatial discretizations or sequential selections. The efficiency of these optimal procedures is tested under actual field conditions. The results show that the optimal MPV is an effective tool for adjustment of existing sampling networks.

Algorithms↗
PubMed2001

Cobalt(II)-selective membrane electrode based on a recently synthesized benzo-substituted macrocyclic diamide.

A PVC-membrane electrode based on a recently synthesized 18-membered macrocyclic diamide is presented. The electrode reveals a Nernstian potentiometric response for Co2+ over a wide concentration range (2.0 x 10(-6)-1.0 x 10(-2) M). The electrode has a response time of about 10 s and can be used for at least 2 months without any divergence. The proposed sensor revealed very good selectivities for Co2+ over a wide variety of other metal ions, and could be used over a wide pH range (3.0-8.0). The detection limit of the sensor is 6.0 x 10(-7) M. It was successfully applied to the direct determination and potentiometric titration of cobalt ion.

Amides↗
PubMed2000

Characterization of conditions required for X-Ray diffraction experiments with protein microcrystals.

The x-ray exposure at which significant radiation damage occurs has been quantified for frozen crystals of bacteriorhodopsin. The maximum exposure to approximately 11-keV x-rays that can be tolerated for high-resolution diffraction experiments is found to be approximately 10(10) photons/microm(2), very close to the value predicted from limits that were measured earlier for electron diffraction exposures. Sample heating, which would further reduce the x-ray exposure that could be tolerated, is not expected to be significant unless the x-ray flux density is well above 10(9) photons/s-microm(2). Crystals of bacteriorhodopsin that contain approximately 10(11) unit cells are found to be large enough to give approximately 100 high-resolution diffraction patterns, each covering one degree of rotation. These measurements are used to develop simple rules of thumb for the minimum crystal size that can be used to record x-ray diffraction data from protein microcrystals. For work with very small microcrystals to be realized in practice, however, it is desirable that there be a significant reduction in the level of background scattering. Background reduction can readily be achieved by improved microcollimation of the x-ray beam, and additional gains can be realized by the use of helium rather than nitrogen in the cold gas stream that is used to keep the protein crystals frozen.

Bacteriorhodopsins↗
PubMed1998

Effects of muscimol or homotaurine on sleep-wake states in alcohol-dependent rats during withdrawal.

Sleep-wake states were studied following withdrawal in 36 adult male wistar alcohol-dependent rats, after chronic administration of ethanol (10 g/kg/24 h) for 13 days. In the light phase of the withdrawal day, 12 alcohol-dependent rats received muscimol (0.25 mg/kg), 12 received homotaurine (140 mg/kg), and 12 received 0.9% physiological saline (10 ml/kg). The results have been compared with a control group of 36 rats that received water during the treatment phase of the experiment, and the 14th day received intraperitoneal muscimol or homotaurine. Muscimol significantly improves the alterations of sleep-wake states in alcohol-withdrawn rats, decreasing the percentage of active wakefulness and increasing the percentage of REMS, but without any action on the latency of appearance of REMS, which remains shortened. The effects of homotaurine are less important on the wakefulness, but it also increases the percentage of REMS without influencing its latency of appearance. The influence of these GABA(A) agonists is not identical during the whole period of survey in the light phase, as there are important differences in the temporal sequences for each of them. We conclude that the stimulation of GABA(A) receptors, of which the activity is decreased during alcohol withdrawal, significantly improves the disturbances in the sleep-wake states in the alcohol-dependent rats, in a time-related manner, and there are significant pharmacodynamic differences between muscimol and homotaurine.

Alcoholic Intoxication↗
PubMed1997

Stability of brain content of magnesium in experimental hypomagnesemia.

Magnesium is important in cerebral function. If there is a deficiency and neurological symptoms accrue, we hypothesised that Mg2+ deficiency causes neurological symptoms by decreasing the level of Mg2+ in cerebral tissue. The content of magnesium was determined in 12 brain structures in magnesium-deficient rats. Experiments were carried out for 40 days in two groups of Wistar male rats made magnesium-deficient (MD) by a well-controlled diet (50 mg of Mg2+/kg of food), and a control group (CG) rats fed normal diet (1 g of Mg2+/kg of food). At the end of the 40 days, the clinical signs of hypomagnesemia were sought in the MD rats and Mg2+ concentration levels were measured in the blood and brain. The results showed variable distribution of Mg2+ in the different brain structures, both in CG and MD rats; in the MD rats there is an important stability of global Mg2+ content of the brain. Although the global values for Mg2+ in the brain did not decline in MD rats, there was a significant decrease in Mg2+ in the brainstem. We conclude that the brain is able to maintain a stable concentration of Mg2+ during chronic hypomagnesemia, but its topographic variations could account for some of neurological signs accompanying this condition.

Animals↗
PubMed1996

Secondary structure of the outer membrane proteins OmpA of Escherichia coli and OprF of Pseudomonas aeruginosa.

When purified without the use of ionic detergents, both OmpA and OprF proteins contained nearly 20% alpha-helical structures, which disappeared completely upon the addition of sodium dodecyl sulfate. This result suggests that the proteins fold in a similar manner, with an N-terminal, membrane-spanning beta-barrel domain and a C-terminal, globular, periplasmic domain.

Bacterial Outer Membrane Proteins↗
PubMed1992

Effects of alcohol dependence on shock-induced fighting: action of muscimol and homotaurine.

We have applied the electroshock-induced fighting behavior to the study of experimental alcohol dependence. Adult Wistar rats were intoxicated chronically with ethanol (10 g/kg/24 h) for 13 days. Electroshock-induced fighting behavior was studied during chronic intoxication and withdrawal in comparison with normal rats receiving a water-carbohydrate solution isocaloric to ethanol. Rats were divided into groups receiving respectively muscimol (0.25 mg/kg), a GABAA agonist; homotaurine (140 mg/kg) a GABA mimetic; and physiological saline (10 ml/kg), intraperitoneally. During chronic intoxication, rats showed an increase in defensive-fighting behavior. Withdrawal accentuated the aggressive behavior and muscimol and homotaurine inhibited it. These results confirm the relevance of the electroshock-induced defensive fighting behavior test in chronic intoxication with alcohol, but to show the involvement of GABAergic transmission in the behavioral effects of alcohol withdrawal, additional experiments with other GABA mimetics and with GABA antagonists should be considered.

Aggression↗
PubMed1990

Circadian variations in vigilance states in the alcohol-dependent rat.

Waking and sleep states were studied in the alcohol-dependent rat after administration of ethanol (416 mg/kg/hr) by indwelling intragastric catheter (IGC) for 13 days. Electropolygraphic recordings performed for a total of 24 hr from the start of withdrawal were compared with those of control rats receiving water by IGC and showed 1) that rapid eye movement sleep was the most sensitive of the four vigilance states studied. A decrease was noted both for the total duration of recording and for the light period; 2) that nonactive wakefulness was the only vigilance state to show an inversion of percentages between the light and dark period; 3) that the light period was the best time for studying changes in vigilance states. Changes included increased percentages of active and nonactive wakefulness and decreased percentages of slow-wave and rapid eye movement sleep. This was due to a change in the number of episodes rather to a change in their mean duration. No significant change occurred during the dark period.

Alcohol Withdrawal Delirium↗
PubMed1987

The frequency of shifts between alternative equilibria.

We derive a formula giving the frequency with which random drift shifts a population between alternative equilibria. This formula is valid when such shifts are rare (Ns much greater than 1), and applies over a wide range of mutation rates. When the number of mutations entering the population is low (4 N mu much less than 1), the rate of stochastic shifts reduces to the product of the mutation rate and the probability of fixation of a single mutation. However, when many mutations enter the population in each generation (4 N mu much greater than 1), the rate is higher than would be expected if mutations were established independently, and converges to that given by a gaussian approximation. We apply recent results on bistable systems to extend this formula to the general multidimensional case. This gives an explicit expression for the frequency of stochastic shifts, which depends only on the equilibrium probability distribution near the saddle point separating the alternative stable states. The plausibility of theories of speciation through random drift are discussed in the light of these results.

Adaptation, Biological↗
PubMed1984

Vigilance states and cerebral monoamine metabolism in experimental magnesium deficiency.

Correlations between cerebral monoamine metabolism and electrophysiological parameters were compared in 18 male Wistar rats subjected to a magnesium-restricted diet and in 14 normal rats. During the 40-day experimental period, plasma and erythrocyte Mg2+ levels and plasma Ca2+ and phosphorus levels were measured, and electroencephalographic tracings as well as clinical data were recorded at regular intervals. At the end of the study, the animals were killed and cerebral concentrations of monoamines and their metabolites were determined. Cerebral monoamine disorders characterized by a rise of dopamine and 5-hydroxyindole-3-acetic acid levels were observed in the magnesium-deficient group. This rise was accompanied by an increase in wakefulness and a decrease in sleep percent. The role played by magnesium in cerebral monoamine metabolism and sleep cycles is discussed.

3,4-Dihydroxyphenylacetic Acid↗
PubMedDate not supplied

EEG effects of a single low dose of ethanol on afternoon sleep in the nonalcohol-dependent adult.

Three polygraphic recordings (PGR) of afternoon sleep (ANS) related to the duration of one sleep cycle, i.e., 90 min, were performed in 14 healthy adult volunteers (7 men and 7 women): two reference PGR, on two consecutive days (before ingestion of alcohol). Only the second being retained: reference PGR = P1; another recording, on day 3, 50 min after the start of single slow oral ingestion of the equivalent of 0.25 g 95% ethyl alcohol (ETOH) per kg body weight. Alcohol was ingested as 40 degrees whiskey, and the volume administered ranged from 34.5 to 66 ml (ETOH polygraphic recording = P2). Analysis of polygraphic traces was carried out according to the criteria of Rechtschaffen and Kales, and results were presented using the parameters adopted by Gross et al. (2). A single low dose of alcohol, leading to a low mean blood alcohol level (below 30 mg/100 ml, range 9 to 29 mg/100 ml), clearly perturbs sleep in the normal nonalcohol-dependent adult. In this context, ETOH does not appear to be a hypnotic since: a) the latencies to onset of sleep and the appearance of stages II, III, and IV of slow-wave sleep (SWS) are not shortened; b) the total duration of sleep, the percentage of delta sleep, and the duration (and percentage) of rapid eye movement sleep (REMS) are decreased; c) the number, duration, and percentages of intrasleep awakenings are increased, as are the number of stage changes. In addition, the study of afternoon sleep has shown itself to be a sensitive and reliable test for the analysis of the effects of a low dose of alcohol on nonalcohol-dependent subjects.

Adult↗