Dye-surfactant interaction: modulation of photophysics of an ionic styryl dye.

The interesting modulation of multibond rotation-induced intramolecular charge transfer photophysics of 2-(4-(dimethylamino) styryl)-1-methylpyridinium iodide in different micelles due to different contributions of twisted intramolecular charge transfer (TICT) and hydrogen bonding deactivation channels have been reported in this paper. 2-(4-(dimethylamino) styryl)-1-methylpyridinium iodide enters into all the micelles in different positions from the water solution due to active hydrophobic force and electrostatic field, as revealed from the shift and intensity of intramolecular charge transfer (ICT) band. The presence of mechanically trapped water with the addition of salt and inherent thermodynamic water controls the ICT emission. Analysis of spectral data before and after the addition of salt confirms the orientation of 2-(4-(dimethylamino) styryl)-1-methylpyridinium iodide in cationic and anionic micelles.

Orientational dynamics of a charge transfer complex in cyclodextrin cavity as receptor.

This paper reports the structure and dynamics of a twisted intermolecular charge transfer molecule 2-(4-(dimethylamino) styryl)-1-methylpyridinium iodide (o-DASPMI) included inside alpha-, beta- and gamma-cyclodextrin, investigated by using steady state and time-resolved emission spectroscopy and also theoretical modeling. A nice 1 : 1 inclusion complex with beta-CD in the excited state could be found with the dimethylamino group of the molecule sticking out as revealed from steady state and time-resolved emission. The inclusion complex has a longer decay time compared to that in neat water. Time-resolved anisotropy decay has been used to study the rotational dynamics of the molecule inside cyclodextrin cavity. The average angular structure of the inclusion complex as found from semiempirical PM3 calculations corroborates excellently the experimental results of angular orientation in beta-CD. The minimum energy of the complex is found to be nearly 5 A in the length of the molecule with the dimethylamino part sticking out in the bulk water. Hydrogen bonding at the rim hinders the inclusion complex of o-DASPMI in gamma-CD and instead it produces association at the rim. Hydrogen bond breaker urea breaks the bonding of o-DASPMI with the rim of gamma-CD and the formation of inclusion complex with gamma-CD ensues.

Uncomplicated falciparum malaria complicated by salmonella septicaemia: cause not coincidence.

A male patient of 10-year-old presented with fever, headache and vomiting for last few days. He was being treated with antimalarial drugs. On 14th day of illness he again showed rise of temperature. His blood culture showed growth of Salmonella typhi. He was treated with ceftriaxone and responded favourably. Here uncomplicated falciparum malaria developed a secondary infection with salmonella during hospital stay. This uncommon association was noted rather than a mere coincidence, which rarely reported in literature.

Stress response in infants undergoing cardiac surgery: a randomized study of fentanyl bolus, fentanyl infusion, and fentanyl-midazolam infusion.

UNLABELLED: There have been significant changes in the management of neonates and infants undergoing cardiac surgery in the past decade. We have evaluated in this prospective, randomized, double-blinded study the effect of large-dose fentanyl anesthesia, with or without midazolam, on stress responses and outcome. Forty-five patients < 6 mo of age received bolus fentanyl (Group 1), fentanyl by continuous infusion (Group 2), or fentanyl-midazolam infusion (Group 3). Epinephrine, norepinephrine, cortisol, adrenocortical hormone, glucose, and lactate were measured after the induction (T1), after sternotomy (T2), 15 min after initiating cardiopulmonary bypass (T3), at the end of surgery (T4), and after 24 h in the intensive care unit (T5). Plasma fentanyl concentrations were obtained at all time points except at T5. Within each group epinephrine, norepinephrine, cortisol, glucose and lactate levels were significantly larger at T4 (P values < 0.01), but there were no differences among groups. Within groups, fentanyl levels were significantly larger in Groups 2 and 3 (P < 0.001) at T4, and among groups, the fentanyl level was larger only at T2 in Group 1 compared with Groups 2 and 3 (P < 0.006). There were no deaths or postoperative complications, and no significant differences in duration of mechanical ventilation or intensive care unit or hospital stay. Fentanyl dosing strategies, with or without midazolam, do not prevent a hormonal or metabolic stress response in infants undergoing cardiac surgery. IMPLICATIONS: We demonstrated a significant endocrine stress response in infants with well compensated congenital cardiac disease undergoing cardiac surgery, but without adverse postoperative outcome. The use of large-dose fentanyl, with or without midazolam, with the intention of providing "stress free" anesthesia, does not appear to be an important determinant of early postoperative outcome.

Prolonged mivacurium infusion in young and elderly adults.

PURPOSE: This study was designed to evaluate pharmacodynamically and pharmacokinetically if the cis-cis isomer of mivacurium contributed to neuromuscular block during prolonged infusions lasting more than four hours in young adult and elderly (> 60 yr) patients. METHODS: The mechanomyogramic neuromuscular response of the adductor pollicis was recorded in 32 adults 18-59 yr. and 19 elderly (> 60 yr.) patients during N2O:O2:opioid anaesthesia. The mivacurium infusion rate was adjusted to maintain single twitch depression at 95 +/- 4% of control. Blood samples were taken every 30 min to determine the plasma concentration of cis-cis isomer of mivacurium. At the end of the surgical procedure, patients were allowed to recover spontaneously to at least 25% of control twitch response. RESULTS: The mean mivacurium infusion requirement to maintain 97 +/- 1 (mean +/- SD)% depression of the twitch response was 6.0 +/- 0.4 micrograms.kg-1.min-1 in young adults, and 4.3 +/- 0.3 micrograms.kg-1.min-1 in elderly patients (P < 0.001). The infusion requirement in patients with low plasma cholinesterase activity was the lowest 2.4 +/- 1.2 micrograms.kg-1.min-1. Plasma cis-cis isomer concentrations reached peak levels within one-two hours and remained relatively constant throughout the duration of infusion even in patients with low cholinesterase activity. There was no relationship between duration of infusion, plasma concentrations of cis-cis isomer and the early recovery indices of mivacurium (up to 25%). Neuromuscular transmission recovered adequately with or without antagonism in all patients. CONCLUSION: When the mivacurium infusion was titrated to maintain 95 +/- 4% twitch depression, the plasma concentration of the cis-cis isomer did not increase during prolonged infusions (four hours) and neuromuscular transmission recovers satisfactorily.

Recovery from doxacurium infusion administered to produce immobility for more than four days in pediatric patients in the intensive care unit.

Doxacurium was administered by titrated infusion to 14 pediatric patients for 4.7-12.3 days after laryngotracheal reconstruction to produce minimum spontaneous movement and less than five posttetanic movements of the first toe after stimulation of the posterior tibial nerve. Recovery was documented by stimulation of the ulnar nerve with 2 Hz for 2 s (train-of-four [TOF]) at intervals of 1 min and measurement of the ratio of the fourth to the first response (TOF ratio) at the adductor pollicis. During spontaneous recovery, the TOF ratio was between 0.4 and 0.7 for 0.6-3.3 h, mean (SEM) 2.2 (0.31) h. The TOF ratio equaled 1 between 4.7 and 23.0 h, mean (SEM) 11.0 (2.1) h after termination of doxacurium infusion. In six of the patients, weakness and decreased coordination were noted for a few days to weeks postoperatively. There were no complications related to impairment of upper airway function or ventilation in those patients who had recovery of neuromuscular transmission to the extent of TOF ratio equal to 1 prior to extubation or in those patients in whom weakness or lack of coordination was noted after tracheal extubation.

Fentanyl and alfentanil plasma protein binding in preterm and term neonates.

The effects of gestational age (GA) and plasma protein concentrations on the plasma protein binding of fentanyl and alfentanil were studied in preterm and term neonates. Binding experiments were performed using split-cell equilibrium dialysis. Fentanyl and alfentanil concentrations were measured using specific radioimmunoassay, and the proteins albumin and alpha-1-acid glycoprotein (AAG) were measured using radial immunodiffusion assays. In the preterm neonates, 77% of fentanyl and 65% of alfentanil was bound. In the term neonates, 70% of fentanyl and 79% of alfentanil was bound. The binding ratio of alfentanil showed a positive correlation with gestational age and AAG concentration. The binding ratio of fentanyl showed a weak, negative correlation with gestational age. These data indicate that fentanyl and alfentanil are not interchangeable at the GA studied because of age-related changes in protein binding.

Acidic and basic fibroblast growth factor mRNA and protein in mouse mammary glands.

Previous studies suggest that members of the fibroblast growth factor (FGF) family are mitogenic to mammary epithelium. In order to determine expression of acidic and basic FGF (aFGF and bFGF) during mammary development, mice were euthanized as virgins, early pregnant, mid-pregnant, late-pregnant, or during early lactation. Mammary expression of both aFGF and bFGF mRNA increased through pregnancy. Acidic FGF mRNA continued to increase during early lactation, but basic FGF message level decreased drastically during early lactation. Western blots probed with anti-aFGF showed four immunoreactive bands approx 30, 48, 52, and 55-kDa in size. The 30-, 48-, and 55-kDa bands for aFGF were expressed at low levels during virgin and early pregnant stages but were more prominent during the later stages. The 52-kDa band was high during the virgin and early pregnant stages and low in mid-pregnancy through early lactation. Blots probed with anti-bFGF showed two bands approx 30 and 50 kDa in size. Both bands increased through early-pregnancy, but during late-pregnancy there was a decrease in immunoreactive protein levels, which remained low during early lactation. Experiments to determine where FGF mRNAs are produced in the mammary gland suggest that both FGFs may be produced in the stroma, leading to the hypothesis that aFGF and bFGF are stromally produced growth factors and probably act on the epithelial component of the gland in a paracrine fashion.

Hormonal regulation of acidic and basic fibroblast growth factor production and expression in mouse mammary gland.

Based on previous results showing developmental regulation of aFGF and bFGF, we evaluated the effect of various hormones on aFGF and bFGF mRNA levels, in mammary gland. Northern blots indicated that estrogen alone increased aFGF but had no effect on bFGF messenger RNA level. Progesterone alone increased aFGF and bFGF mRNA levels. Estrogen and progesterone together increased aFGF mRNA level in mammary gland, but the increase was no greater than that caused by either hormone alone. However, the combination of estrogen and progesterone had no effect on bFGF message level. PRL or GH, when administered with estrogen and progesterone, increased aFGF, but did not have any effect on bFGF message level. However, when PRL and GH were administered together with estrogen and progesterone, they increased bFGF messenger RNA level. Ovarian steroid withdrawal increased aFGF, but did not have any effect on bFGF mRNA accumulation. PRL alone (in the absence of estrogen and progesterone) decreased aFGF, whereas it had no effect on bFGF message level. Hydrocortisone alone decreased aFGF, but increased bFGF mRNA level. However, PRL and hydrocortisone increased aFGF, but did not have any significant effect on bFGF message level. In the overall model, during growth of the mammary gland, ovarian steroids cause an increase in aFGF mRNA. During lactogenesis, ovarian steroid withdrawal causes an increase in aFGF messenger RNA levels. Lactogenic hormones together cause a further increase in aFGF message levels.

Photophysics and photochemistry of new verdin compounds.

Five different verdins, including one zinc metal chelate, were examined by laser flash techniques. Triplet molar absorption coefficients, triplet and singlet oxygen quantum yields and triplet lifetimes were determined. Zinc methyl pyroverdin (ZNMPV), copro II verdin trimethyl ester (CVTME) and deuteroverdin methyl ester (DVME) have the highest triplet and singlet oxygen quantum yields. ZNMPV and CVTME have the longest triplet lifetimes. Our data are consistent with singlet oxygen as the primary modality for phototherapy and it is suggested that DVME and CVTME may be useful agents.

The photosensitizers benzophenoxazine and thiazines: comprehensive investigation of photophysical and photochemical properties.

Eight differently substituted title dye compounds have been investigated regarding intersystem crossing, triplet state, fluorescence and singlet excited state pKa properties. In general, non-halogenated oxazines and thiazines as well as a mono bromooxazine show very low triplet quantum yields, phi tau (less than 0.03) and relatively long triplet lifetimes (approximately 40 microseconds) in acidic methanol. The phi tau data correlate well with known singlet oxygen yields. In basic methanol no triplet transient is observed but a significant yield of a ground state transient protonated (base dye) form is produced with a short lifetime, approximately 400 ns. Fluorescence can be seen simultaneously from both the excited base and the protonated base dye forms in basic methanol. For iodinated oxazine or thiazines, the triplet yield increases and can be as high as 0.5 (diiodo case) in acidic methanol. The triplet lifetimes are further shortened to approximately 10 microseconds compared to the non-iodinated derivatives above. The triplet yields of the iodo compounds are higher or equal to known singlet oxygen yields. In basic methanol triplet yields up to 0.2 can be seen, the triplet lifetime are shortened still further to 1 microsecond but no observable protonated form is produced (in distinction to the non-iodinated cases). Consideration is given to the correlation of triplet and singlet oxygen yields, ground and excited pKa properties, spin-orbit coupling and internal conversion properties, solvent effects, and phototherapeutic activity of these dyes.

Photochemical and photophysical properties of piroxicam and benoxaprofen in various solvents.

Laser flash spectroscopy was used to examine the title compounds. Piroxicam has a triplet transient with a maximum near 450 and a lifetime of 3-21 microseconds depending on the solvent. The relative quantum yield is highly solvent dependent being maximum in toluene and greater than or equal to 14 fold lower in hydrogen bonding solvents. There is another transient which is assigned as a proton transferred ground state transient. Some permanent photoproduct also appears to be produced. Benoxaprofen also has a triplet transient with a maximum near 420 nm with a lifetime of 65 microseconds to greater than or equal to 250 microseconds depending on the solvent. In this case, the relative quantum yield only slightly varies among polar and hydrogen bonding solvents. This is in marked contrast to published data on the fluorescence yield. Some permanent photoproduct appears to be produced.

In vitro metabolism of mivacurium chloride (BW B1090U) and succinylcholine.

The in vitro rates of metabolism of mivacurium chloride and succinylcholine in pooled human plasma were compared. In addition, the rate of metabolism of mivacurium in buffered solutions of butyrylcholinesterase (E.C. 3.1.1.8) and acetylcholinesterase (E.C. 3.1.1.7) was determined. Succinylcholine concentrations were measured spectrophotometrically, and mivacurium concentrations were determined with a high-pressure liquid chromatographic assay. The hydrolysis of mivacurium in plasma followed first-order kinetics, and the rate of hydrolysis decreased as plasma was serially diluted. The Michaelis-Menten constant (Km) for mivacurium metabolism in plasma was 245 mumol/L, and the maximum velocity (Vmax) was 50 U/L; the Km for succinylcholine was 37 mumol/L, and Vmax was 74 U/L. At comparable multiples of the Km the hydrolysis rate of mivacurium was 70% of that of succinylcholine. Mivacurium was metabolized significantly in solutions containing butyrylcholinesterase, but only minimally in solutions containing acetylcholinesterase.

Cimetidine does not inhibit plasma cholinesterase activity.

Cimetidine increases the duration of action of succinylcholine several-fold by an unknown mechanism. The hydrolysis rate of succinylcholine by human plasma was measured with a modified spectrophotometric assay. At a concentration of 1-50 micrograms/ml cimetidine did not inhibit the hydrolysis of succinylcholine. It is concluded that cimetidine may have an effect at the neuromuscular junction but does not inhibit plasma cholinesterase.

Pharmacokinetics of atracurium in anaesthetized infants and children.

The pharmacokinetics of atracurium were studied in infants and children anaesthetized with isoflurane and nitrous oxide in oxygen. There were no significant differences in volume of distribution (area) (139 v. 152 ml kg-1), clearance (5.1 v. 5.3 ml kg-1 min-1), T1/2 alpha (2.1 v. 2.0 min), or T1/2 beta (19.1 v. 20.3 min) between children with normal hepatic and renal function and those with moderately impaired hepatic function presenting for hepatic transplantation. There were significant differences in volume of distribution (area) (176 v. 139 ml kg-1) and in clearance of atracurium (9.1 v. 5.1 ml kg-1 min-1) between infants and children with normal excretory function. In infants the clearance of atracurium in ml m-2 min-1 (153 v. 133) tended to be greater and the T1/2 alpha and T1/2 beta tended to be shorter (1.0 v. 2.0 and 13.6 v. 19.1) than in children with normal excretory function; however, these trends did not reach statistical significance. Plasma laudanosine concentration was around 100 ng ml-1 greater in patients with liver disease than in normal children from 15-45 min following a bolus of atracurium 0.5 mg kg-1.

In vitro degradation of atracurium in human plasma.

The degradation of atracurium and the formation of laudanosine was examined in vitro in both Sorensen buffer and human plasma using sensitive, specific high pressure liquid chromatographic assays to determine drug concentrations. At normal physiological pH and temperature, the degradation of atracurium was threefold more rapid in plasma than in buffer. Laudanosine is the major end-product of atracurium degradation in buffer or in plasma; its production is more rapid in plasma than in buffer. Dilution of plasma constituents or the use of diisopropylfluorophosphate (a potent esterase inhibitor), slows the degradation of atracurium and the production of laudanosine. We conclude that, although ester hydrolysis is the major metabolic pathway of atracurium degradation, Hofmann elimination provides a "safety net" in its clinical use.

Further studies on the activation of glucocerebrosidase by a heat-stable factor from Gaucher spleen.

Using Sephadex G-75 and DEAE-cellulose column chromatography, an 8270-Da glycopeptide (designated Fragment II) has been isolated from a cyanogen bromide-formic acid digest of a heat-stable factor from Gaucher spleen which activates a lipid-depleted preparation of lysosomal glucocerebrosidase from human liver. Fragment II contains all of the activity present in the native heat-stable factor. Compared with the parent factor, Fragment II contains four fewer cysteine and methionine residues and one less of each of the following: aspartic acid, threonine, serine, valine, isoleucine, and leucine. Nearly all of the monosaccharides present in the parent heat-stable factor can be accounted for in Fragment II, including three glucosamine, three mannose, one sialic acid, and one fucose. By itself, Fragment II has little or no stimulatory activity; its major effect is to markedly increase the sensitivity of glucocerebrosidase to activation by phosphatidylserine. A mixture of 1 microgram phosphatidylserine and 2 micrograms of the cyanogen bromide fragment activates the lipid-depleted preparation of glucocerebrosidase 50% more than 30 micrograms phosphatidylserine alone. Analysis of the Km and Vmax of glucocerebrosidase at various hydrogen ion concentrations revealed that the heat-stable factor and phosphatidylserine together dramatically increase the catalytic efficiency (Vmax/Km) of glucocerebrosidase while making apparent three ionizable groups that participate in the catalysis. Phosphatidylserine alone recruits two ionizable groups, but catalytic efficiency is lower than when heat-stable factor is also present. Heat-stable factor alone has no discernable effect on the ionization of functional groups on the enzyme or on catalytic efficiency. By sucrose density gradient ultracentrifugation, it was shown that preincubation of rat liver glucocerebrosidase with phosphatidylserine and heat-stable factor shifted the enzyme completely from a 56,600-Da form to a 188,100-Da form. The activity of the slower sedimenting form of glucocerebrosidase was totally dependent upon exogenous bile salt activator, whereas the faster sedimenting form exhibited the same activity in the presence or absence of sodium taurocholate. It appears that the heat-stable factor promotes the transfer of phosphatidylserine to glucocerebrosidase, which, in turn, results in an increase in both the catalytic efficiency and size of the enzyme.

Analysis of data from multiclinic experiments.

The analysis of data from multiclinic experiments is put in the framework of a mixed model in which the treatment effects are assumed to be fixed, and clinic and clinic by treatment effects are assumed to be random. The analysis in the unbalanced case is developed under the usual mixed model assumptions. The maximum likelihood estimates and likelihood ratio tests of the parameters are derived following the approach of Hemmerle and Hartley (1973). The T2 test is shown to be asymptotically equivalent to the analysis of variance F test in the balanced case. The distribution of the test statistic in the case of proportional cell frequencies is also discussed. Some investigation is made of the distribution of the test statistic by applying the method of Box (1954).