Determination of citalopram enantiomers in human plasma by liquid chromatographic separation on a Chiral-AGP column

A liquid chromatographic method for the quantitative analysis of S-(+)- and R-(−)-citalopram in human plasma has been developed and validated. The enantiomers of citalopram and the internal standard, R-(+)-propranolol, were extracted from alkaline plasma with 2% n-butanol in n-hexane. After a clean-up step, the organic phase was evaporated and the residues dissolved in 50–100 μl of 0.001 M HCl. The separation was performed on a Chiral-AGP column with 3.0 mM N-dodecyl-N,N-dimethylammonio-3-propanesulfonate and 10 mM hexanoic acid in phosphate buffer pH 6.5 as the mobile phase. The limit of detection was estimated to be 1 ng/ml (S/N≈3) for each enantiomer monitoring UV absorption at 240 nm. In the range studied, 2.31–191 ng/ml, the recoveries were quantitative and the coefficients of variations were between 2.47% and 11.5%.     

Effect of temperature on the reversal of the calcium ion pump in sarcoplasmic reticulum.

Sarcoplasmic-reticulum vesicles were actively loaded with Ca2+ in the presence of phosphate, and the ADP-induced Ca2+ efflux and ATP synthesis were measured as a function of temperature. Arrhenius plots show break points for both processes at about 18 and 37 degrees C. Between 18 and 37 degrees C, Ca2+ efflux and ATP synthesis occur with an activation energy of 67.2-71.4 kJ/mol, whereas it is about 189-210 kJ/mol for temperatures below 18 degrees C. Above 37 degrees C, the rates of ADP-induced Ca2+ release and of ATP synthesis sharply decline until the temperature reaches about 42 degrees C. Above this temperature, the Ca2+ efflux increases again even in absence of ADP, although the synthesis of ATP is inhibited, which reflects leakiness of the vesicles. The results show that the transition temperatures for ADP-induced Ca2+ efflux and for ATP synthesis resemble those for active Ca2+ uptake, which indicates that the same coupling mechanism is involved during the inward and outward Ca2+ translocations across the membrane.     

Effect of temperature on glycerol retention in the halotolerant algae dunaliella and asteromonas

Algae of the genera Dunaliella and Asteromonas can maintain extremely high concentration gradients (>10⁴) of glycerol between the intracellular space and the medium. This unique ability is highly temperature-dependent. Treating the algae for several minutes at temperatures exceeding 60 C causes complete release of all the internally held glycerol; 50% release occurs around 50 C, but essentially none is released below 40 C. A similar behavior was observed in several species of Dunaliella, and one of Asteromonas and is independent of the salt concentration of the medium. The underlying mechanism may involve a temperature-dependent conformational transition of a component of the cellular membrane which is essential for glycerol impermeability.     

Reversal of elution order for profen acid enantiomers in packed-column SFC on Chiralpak AD

Enantiomeric separations of four 2-substituted propionic acid drugs have been studied using packed-column supercritical fluid chromatography (SFC) with amylose tris(3,5-dimethylphenylcarbamate) coated on silica as support (Chiralpak AD). Under standard conditions (i.e., flow rate, 1.5 ml/min; column temperature, 30 degrees C; back-pressure, 150 bar), the order of elution could be reversed when the polar alcohol modifier methanol in carbon dioxide was replaced by 2-propanol for ibuprofen, ketoprofen, and naproxen. For flurbiprofen, with the highest selectivity factor, no reversal was observed, although selectivity was reduced significantly with higher alcohols. Naproxen and flurbiprofen were also investigated with 2-butanol and 2-pentanol. The former showed reversal of elution order but not the latter. For higher alcohol modifiers, including 2-propanol, the peak symmetry was poor but could be improved by addition of citric acid in the alcohol modifier. These results stress the importance to investigate enantiomer elution order during the development of enantioselective methods and when chromatographic conditions are optimized. Preliminary experiments with column temperatures over the range of -15 to 45 degrees C revealed that, in a few cases, reversal took place with a change in temperature only.     

Effect of butaclamol enantiomers on hypothalamic tyrosine hydroxylase in the rat brain

The authors demonstrate stereospecificity of the action of butaclamol enantiomers on substrate inhibition of hypothalamic tyrosine hydroxylase (TH) and regulation of the tyrosine hydroxylase response by the presynaptic membrane (presynaptic receptors) of rat hypothalamus synaptosomes under membrane activation with dopamine. The effect of (+)-butaclamol on the substrate inhibition of TH was noticeable at a concentration of 10(-8)M, reaching a maximum at 10(-5)M. (-)-Butaclamol administered at the same concentrations did not influence the substrate inhibition of the enzyme. (+)-Butaclamol added to the incubation medium containing hypothalamic synaptosomes concurrently with dopamine (10(-5)M) completely blocked the regulatory action of the latter on TH, with this action mediated via presynaptic receptors. (-)-Butaclamol (10(-5)M) antagonized the action of dopamine under the same conditions. The data obtained indicate high stereo-specificity of butaclamol enantiomers as regards their effect on presynaptic regulation of TH, suggesting that elimination of the substrate inhibition of hypothalamic TH is a stereoselective effect of neuroleptics and can be a prognostically important criterion in the appraisal of compounds with potential neuroleptic activity.     

Dominance order reversal between two groups of free-ranging rhesus monkeys

A rank reversal between two groups of free-ranging rhesus monkeys (Macaca mulatta) was observed.The group which became dominant through the reversal had been transplanted from another free-ranging colony eight months before. The actions of peripheral adult males of this group were important in effecting the reversal. Support of the peripheral males by the rest of the group was crucial in effecting the reversal.The development of the reversal indicates that social tradition and individual relationships between monkeys are important in maintaining and establishing intergroup rank.     

Induced reversal of order of cell division and DNA replication in Tetrahymena.

The sequence of macronuclear DNA replication (S) and cell division (D) in two normal or synchronized cell cycles in (amicronucleate) Tetrahymena can be symbolized S₁, D₁, S₂, D₂. Using heat shock synchronized cells, the final heat shock (33.8 °C) has been extended from 20 min to 5 h. This leads to an altered sequence: S₁, S₂, D₁, D₂, characterized by reversal of D₁ and S₂. S₂ occurs during the extended shock, D₁ comes in standard time after the shock has been discontinued. Thus prolonged stay at elevated temperature can dissociate two cell cycles into a subcycle of DNA replication followed by a subcycle of cell division. S₁ and S₂ charge the cells with four times the amount of DNA in newly divided cells, and D₁ and D₂ partition this DNA to four cells. Synthesis of DNA between D₁ and D₂ is not required, and it occurs in only few cells.     

Optimizing the sample size and the retention parameters to achieve maximum production rates for enantiomers in chiral chromatography

The optimum experimental conditions (sample size and mobile phase composition) are calculated for maximum production rate of either one of two enantiomers contained in feeds of different compositions (1/1, 1/10, and 10/1). The products are obtained at 99% purity. The calculations use the equilibrium-dispersive model of chromatography and the equilibrium isotherms determined experimentally from the rear, diffuse boundary of overloaded elution profiles. The production rate measured experimentally under the optimum conditions calculated agree with 4% of the calculated values. There is an optimum value for the retention factor which is higher than predicted by a model assuming constant separation factor, because both separation factor and retention decrease with increasing organic solvent concentration in the mobile phase. (c) 1992 John Wiley & Sons, Inc.     

Geographical and taxonomic variation in the pattern of natural sex reversal in the teleost fish order Synbranchiformes

Two types of males have been found in all members of the Synbranchidae. The primary males are born as males and possess single, but bilobular, testes with two gonadal sinuses. Secondary males evolve by sex reversal of functional females and possess a lamellar organization of the testes with three gonadal sinuses and often ovarian residua. Undifferentiated races lack primary males and are recorded only from certain geographical areas. Rudimentary hermaphroditism is classified as a derived evolutionary stage and gonochorism, which is represented in the Amphipnoidae, a primitive evolutionary condition. The alternative of primary versus secondary male is decided by a genetic shift mechanism instead of by environmental conditions. The precarious poise of sexual identity in the Synbranchidae is defined as a switch mechanism to compensate for the vicissitudes of the natural habitat, allowing reproduction in spite of severe population depletions. Based on an examination of its gonadal structure, Alabes is considered to have no phylogenetic affinities with the Synbranchiformes. The occurrence of natural hermaphroditism in phylogenetically unrelated groups of fishes in radically different environments is regarded as the results of chance evolution, rather than convergence.     

Effect of check size on the pattern reversal visual evoked potential

Modifications of the components of pattern-reversal visual evoked potentials (PR-VEP) with changes in check size of the stimulating pattern were studied in 11 healthy subjects. We made use of 8 different check size ranging between 10 and 90 min of arc. Changes in the check size modified in different manners the latencies and amplitudes of N75, P100 and N145. Two-step statistical analyses using the polynomial regression analysis method revealed significant modifications of latencies of the 3 components, but non-significant modifications of the amplitudes, except for N75. The latency and amplitude of N75 showed a significant inverse linear relationship with the logarithm of the check size, while the P100 and N145 latencies showed significant curvilinear relationships, with minimal latencies at check sizes around 35 min. These findings suggest different physiological properties of N75 from those of P100 and N145, and hence, the necessity to establish normal values for each check size of stimulation, for application in clinical studies.     

Effect of L-arginine on the retention of macrophage tumoricidal activity

It has been reported that the tumoricidal activity of macrophages (M phi) depends on L-arginine and that L-arginine metabolites such as reactive nitrogen intermediates alter M phi physical capacities. The aim of this report is to investigate the dose-related effect of L-arginine on the expression and retention of M phi tumoricidal activity. Cytotoxicity of M phi activated by IFN-gamma plus LPS was detected in the presence of about 0.1 mM or more of L-arginine. This paralleled the NO2- production in the presence, but not in the absence, of L-arginine. On the other hand, activated M phi were destined to die and lost their tumoricidal activity with time in the presence of 0.3 mM or more L-arginine. They retained, however, considerable activity in the absence or presence of 0.15 mM L-arginine. This retention of M phi cytotoxicity was longer when M phi were preactivated by 100 ng/ml than 10 ng/ml of LPS in combination with IFN-gamma. Addition of indomethacin, an inhibitor of prostaglandin production, did not prevent the decay of M phi cytotoxicity but rather facilitated it even in the absence of L-arginine. Regardless of indomethacin, consecutive stimulation with LPS or LPS plus IFN-gamma during culture was effective in maintaining the tumoricidal activity at a high level. In addition, we found that M phi which had lost tumoricidal activity during culture in L-arginine deficient medium could be reactivated by LPS to attack tumor target cells.     

Effect of change in ambient temperature on core temperature during the daytime

In this study, the hypothesis is tested that continuous increases in ambient temperature (T_a) during daytime would give elevated core and skin temperatures, and consequently better thermal sensation and comfort. Rectal temperature (T_re), skin temperatures and regional dry heat losses at 7 sites were continuously measured for 10 Japanese male subjects in three thermal conditions: cond. 1, stepwise increases in T_a from 26 °C at 9 h00 to 30 °C at 18 h00; cond. 2, steady T_a at 28 °C from 9 h00 to 18 h00 and cond. 3, stepwise decreases in T_a from 30 °C at 9 h00 to 26 °C at 18 h00. Oxygen consumption was measured and thermal sensation and comfort votes were monitored at 15 min intervals. Body weight loss was measured at 1 h intervals. While T_re increased continuously in the morning period in any condition, it increased to a significantly greater (p?<?0.05) 36.9?±?0.3 °C at 18 h00 in cond. 1 relative to 36.7?±?0.28 °C in Cond. 2 and 36.5?±?0.37 °C in cond. 3. Better thermal comfort was observed in the afternoon and the evening in Cond.1 as compared with the other 2 conditions. Thus, a progressive and appropriate increase in T_a may induce optimal cycle in core temperature during daytime, particularly for a resting person.     

Effect of raised alveolar pressure on leukocyte retention in the human lung

To determine whether an increase in alveolar pressure delays the passage of leukocytes (WBCs) through the lung by compressing the lung capillaries, we measured the concentration of WBC across the lung in response to a forced expiratory maneuver. In 20 human subjects, blood was sampled from catheters placed in the pulmonary artery (PA) and left ventricle (LV) before, during, and after a forced expiratory maneuver held for greater than or equal to 20 s against an occluded airway. Pressures were recorded at the mouth and from both catheters. A significant fall in LV WBC (P less than 0.01) but not in PA WBC occurred during or immediately after the maneuver in 18 subjects, with a mean maximum decrease of 26 +/- 12% (SD) from base line (range 9-46%). Between 1 and 3 min after the maneuver, there was an increase in LV and PA WBC (P less than 0.01) above base line. The neutrophil and lymphocyte counts showed similar changes, but erythrocyte and platelet counts remained unchanged. The degree of fall in LV WBC correlated closely (r = 0.68, P less than 0.01) with the changes from lung zone 3 to zone 2 and 1 conditions, as determined from the pressure changes. We conclude that WBCs are retained in the lung during a forced expiratory maneuver because of alveolar capillary compression.