Fluctuating dynamics of nematic liquid crystals using the stochastic method of lines.

We construct Langevin equations describing the fluctuations of the tensor order parameter Q(alphabeta) in nematic liquid crystals by adding noise terms to time-dependent variational equations that follow from the Ginzburg-Landau-de Gennes free energy. The noise is required to preserve the symmetry and tracelessness of the tensor order parameter and must satisfy a fluctuation-dissipation relation at thermal equilibrium. We construct a noise with these properties in a basis of symmetric traceless matrices and show that the Langevin equations can be solved numerically in this basis using a stochastic version of the method of lines. The numerical method is validated by comparing equilibrium probability distributions, structure factors, and dynamic correlations obtained from these numerical solutions with analytic predictions. We demonstrate excellent agreement between numerics and theory. This methodology can be applied to the study of phenomena where fluctuations in both the magnitude and direction of nematic order are important, as for instance, in the nematic swarms which produce enhanced opalescence near the isotropic-nematic transition or the problem of nucleation of the nematic from the isotropic phase.

The isotropic-nematic interface with an oblique anchoring condition.

We present numerical and analytic results for uniaxial and biaxial orders at the isotropic-nematic interface within Ginzburg-Landau-de Gennes theory. We study the case where an oblique anchoring condition is imposed asymptotically on the nematic side of the interface, reproducing results of previous work when this condition reduces to planar or homeotropic anchoring. We construct physically motivated and computationally flexible variational profiles for uniaxial and biaxial orders, comparing our variational results to numerical results obtained from a minimization of the Ginzburg-Landau-de Gennes free energy. While spatial variations of the scalar uniaxial and biaxial order parameters are confined to the neighborhood of the interface, nematic elasticity requires that the director orientation interpolate linearly between either planar or homeotropic anchoring at the location of the interface and the imposed boundary condition at infinity. The selection of planar or homeotropic anchoring at the interface is governed by the sign of the Ginzburg-Landau-de Gennes elastic coefficient L(2). Our variational calculations are in close agreement with our numerics and agree qualitatively with results from density functional theory and molecular simulations.

Numerical method of lines for the relaxational dynamics of nematic liquid crystals.

We propose an efficient numerical scheme, based on the method of lines, for solving the Landau-de Gennes equations describing the relaxational dynamics of nematic liquid crystals. Our method is computationally easy to implement, balancing requirements of efficiency and accuracy. We benchmark our method through the study of the following problems: the isotropic-nematic interface, growth of nematic droplets in the isotropic phase, and the kinetics of coarsening following a quench into the nematic phase. Our results, obtained through solutions of the full coarse-grained equations of motion with no approximations, provide a stringent test of the de Gennes ansatz for the isotropic-nematic interface, illustrate the anisotropic character of droplets in the nucleation regime, and validate dynamical scaling in the coarsening regime.

An in silico 3D pharmacophore model of chalcones useful in the design of novel antimalarial agents.

Malaria, the most important of the human parasitic diseases, causes about 500 million infections worldwide and over 1 million deaths every year. The search for novel drug candidates against specific parasitic targets is an important goal for antimalarial drug discovery. Recently the antimalarial activity of chalcones has generated great interest. These compounds are small non-chiral molecules with relative high lipophilicity (clogP approximately 5-7), have molecular weights in the range of 300 to 600 g/mol, and possess in vivo efficacy against both P. berghei and P. yeolii. Preliminary data on our on-going chalcone synthesis project indicate that these compounds are active in vitro against P. falciparum, but are rapidly metabolized in liver microsome assays. Structurally-related compounds not including the enone linker are found to be much more metabolically stable and yet have comparable in vitro efficacy. In this study, we have utilized the efficacy data from an in-house on-going chalcone project to develop a 3D pharmacophore for antimalarial activity and used it to conduct virtual screening (in silico search) of a chemical library which resulted in identification of several potent chalcone-like antimalarials. The pharmacophore is found to contain an aromatic and an aliphatic hydrophobic site, one hydrogen bond donor site, and a ring aromatic feature distributed over a 3D space. The identified compounds were not only found to be potent in vitro against several drug resistant and susceptible strains of P. falciparum and have better metabolic stability, but included one with good in vivo efficacy in a mouse model of malaria.

Antileishmanial and antimalarial chalcones: synthesis, efficacy and cytotoxicity of pyridinyl and naphthalenyl analogs.

The antileishmanial and antimalarial activity of methoxy-substituted chalcones (1,3-diphenyl-2-propen-1-ones) is well established. The few analogs prepared to date where the 3-phenyl group is replaced by either a pyridine or naphthalene suggest these modifications are potency enhancing. To explore this hypothesis, sixteen 3-naphthalenyl-1-phenyl-2-prop-1-enones and ten 1-phenyl-3-pyridinyl-2-prop-1-enones were synthesized and their in vitro efficacies against Leishmania donovani and Plasmodium falciparum determined. One inhibitor with submicromolar efficacy against L. donovani was identified (IC50 = 0.95 microM), along with three other potent compounds (IC50 < 5 microM), all of which were 3-pyridin-2-yl derivatives. No inhibitors with submicromolar efficacy against P. falciparum were identified, though several potent compounds were found (IC50 < 5 microM). The cytotoxicity of the five most active L. donovani inhibitors was assessed. At best the IC50 against a primary kidney cell line was around two-fold higher than against L. donovani. Being more active than pentamidine, the 1-phenyl-3-pyridin-2-yl-2-propen-1-ones have potential for further development against leishmaniasis; however it will be essential in such a program to address not only efficacy but also their potential for toxicity.

Monoclonal anti-galactan IgA J 539 binds intercatenarily to its polysaccharide antigen. Observations on the binding of antibody to a macromolecular antigen.

Highly purified P. Zopfii galactan of mol. wt 2 X 10(5) binds monoclonal IgA J 539 with a Ka of 5.80 X 10(5) M-1 if the polysaccharide concn is expressed in blocks of 30 galactosyl residues. This is the same Ka as found for the antibody and methyl beta (1,6)-beta-D-galactopyranosyltetraoside, the ligand capable of filling the entire combining area of the immunoglobulin. This same polysaccharide precipitates monomeric IgA J 539 on agar-double diffusion. It is concluded that the antibody binds to intercatenary galactosyl residues of the antigen.

Galactofuranosyl groups are immunodominant in Aspergillus fumigatus galactomannan.

Aspergillus fumigatus galactomannan was prepared and its structure partially characterized. Galactofuranosyl groups were immunodominant when this polysaccharide antigen was reacted with antibody raised in rabbits.

Anticryptococcal type D antibodies raised in rabbits.

Antibodies to Cryptococcus neoformans Type D have been raised in rabbits. The partially purified antibody pool was fractionated by affinity-chromatography, and it was shown that antibodies to the capsular polysaccharide of Type D were present in the pool. Neither methyl alpha-D-mannopyranoside, nor D-mannose were inhibitors of the binding of Type D polysaccharide to the antibody-pool. Thus it appears that the serum contains antibody populations which are specific for terminal side-chain carbohydrate moieties of the immunizing antigen.

Malabsorption of water miscible vitamin A in children with giardiasis and ascariasis.

Vitamin A absorption was studied using a water-miscible oral preparation of vitamin A in 19 children ages 1 1/2 to 9 years old with giardiasis and/or ascariasis, both before and after their eradication with appropriate therapy, and in three children without parasites. Marked impairment of vitamin A absorption was noted when administered in a water miscible form in children with 1) combined infection with Giardia lamblia and Ascaris lumbricoides, 2) giardiasis alone, and 3) in a proportion of children with ascariasis alone. In children with both giardiasis and ascarasis eradication of the infections promptly lead to a significant improvement in vitamin A absorption and restored it to normal. Children with giardiasis alone also showed improved vitamin A absorption after therapy. In children with ascariasis alone successful therpay did not lead to a statistically significant improvement.

Molecular electronic properties of a series of 4-quinolinecarbinolamines define antimalarial activity profile.

A detailed computational study on a series of 4-quinolinecarbinolamine antimalarials was performed using the semiempirical Austin model 1 (AM1) quantum chemical method to correlate the electronic features with antimalarial activity and to illuminate more completely the fundamental molecular level forces that affect the function and utility of the compounds. Ab initio (3-21G level) calculations were performed on mefloquine, the lead compound in this series, to check the reliability of the AM1 method. Electron density in specific regions of the molecules appears to play the pivotal role toward activity. A large laterally extended negative potential in the frontal portion of the nitrogen atom of the quinoline ring and the absence of negative potential over the molecular plane are crucial for the potent antimalarials. These electrostatic features are likely to be the modulator of hydrophobicity or lipophilicity of the compounds and, hence, determine their activities. The magnitude of the positive potential located by the hydroxyl hydrogen atom also correlates with potent antimalarial activity. Two negative potential regions occur near the hydroxyl oxygen and piperidyl nitrogen atoms. The two negative potential regions and the positive potential located by the hydroxyl hydrogen atom are consistent with intermolecular hydrogen bonding with the cellular effectors. The present modeling study should aid in efficient designing of this class of antimalarial agents.

Methyl-substituted dispiro-1,2,4,5-tetraoxanes: correlations of structural studies with antimalarial activity.

Two tetramethyl-substituted dispiro-1,2,4,5-tetraoxanes (7,8,15, 16-tetraoxadispiro[5.2.5.2]hexadecanes) 3 and 4 were designed as metabolically stable analogues of the dimethyl-substituted dispiro-1, 2,4,5-tetraoxane prototype WR 148999 (2). For a positive control we selected the sterically unhindered tetraoxane 5 (7,8,15, 16-tetraoxadispiro[5.2.5.2]hexadecane), devoid of any substituents. Tetraoxanes 3 and 4 were completely inactive in contrast to tetraoxanes 2 and 5. We hypothesize that the two inactive tetraoxanes possess sufficient steric hindrance about the tetraoxane ring due to the two additional axial methyl groups to prevent their activation to presumed parasiticidal carbon radicals by inhibiting electron transfer from heme or other iron(II) species. For each of the tetraoxanes 2-4, the tetraoxane and both spirocyclohexyl rings are in a chair conformation and the bond lengths and angles are all quite normal except for the C1-C2 bond which is slightly lengthened. Comparison of the modeled and X-ray structures for tetraoxanes 2-5 reveals that molecular mechanics (MMX and MM3) and 3-21G calculations each gave accurate structural parameters such as bond lengths, bond angles, and dihedral angles. In contrast, semiempirical methods such as AM1 gave poor results.

Structural specificity of chloroquine-hematin binding related to inhibition of hematin polymerization and parasite growth.

Considerable data now support the hypothesis that chloroquine (CQ)-hematin binding in the parasite food vacuole leads to inhibition of hematin polymerization and parasite death by hematin poisoning. To better understand the structural specificity of CQ-hematin binding, 13 CQ analogues were chosen and their hematin binding affinity, inhibition of hematin polymerization, and inhibition of parasite growth were measured. As determined by isothermal titration calorimetry (ITC), the stoichiometry data and exothermic binding enthalpies indicated that, like CQ, these analogues bind to two or more hematin mu-oxo dimers in a cofacial pi-pi sandwich-type complex. Association constants (K(a)'s) ranged from 0.46 to 2.9 x 10(5) M(-1) compared to 4.0 x 10(5) M(-1) for CQ. Remarkably, we were not able to measure any significant interaction between hematin mu-oxo dimer and 11, the 6-chloro analogue of CQ. This result indicates that the 7-chloro substituent in CQ is a critical structural determinant in its binding affinity to hematin mu-oxo dimer. Molecular modeling experiments reinforce the view that the enthalpically favorable pi-pi interaction observed in the CQ-hematin mu-oxo dimer complex derives from a favorable alignment of the out-of-plane pi-electron density in CQ and hematin mu-oxo dimer at the points of intermolecular contact. For 4-aminoquinolines related to CQ, our data suggest that electron-withdrawing functional groups at the 7-position of the quinoline ring are required for activity against both hematin polymerization and parasite growth and that chlorine substitution at position 7 is optimal. Our results also confirm that the CQ diaminoalkyl side chain, especially the aliphatic tertiary nitrogen atom, is an important structural determinant in CQ drug resistance. For CQ analogues 1-13, the lack of correlation between K(a) and hematin polymerization IC(50) values suggests that other properties of the CQ-hematin mu-oxo dimer complex, rather than its association constant alone, play a role in the inhibition of hematin polymerization. However, there was a modest correlation between inhibition of hematin polymerization and inhibition of parasite growth when hematin polymerization IC(50) values were normalized for hematin mu-oxo dimer binding affinities, adding further evidence that antimalarial 4-aminoquinolines act by this mechanism.

Vaccines and antibodies in the prevention and treatment of sepsis.

Antibodies to various core glycolipid antigens have been shown to correlate with survival from Gram-negative sepsis. Recent preclinical data also support efficacy of the anti-core glycolipid antibodies in the treatment of sepsis. Failure of some of the previous clinical trials with anti-core glycolipid antibody was probably due to inadequate levels of antibody in those preparations. Future clinical trials must ensure that sufficient amounts of anti-core glycolipid antibodies are present in the circulation of patients with sepsis.

Predicting mosquito repellent potency of N,N-diethyl-m-toluamide (DEET) analogs from molecular electronic properties.

Specific molecular electronic properties of 30 N,N-diethyl-m-toluamide (DEET) analogs demonstrate functional dependence with their reported duration of protection against mosquito bites, thus providing predictors of insect repellent efficacy. No single electronic property is sufficient to predict repellent efficacy as measured by protection time, rather a set of specific electronic properties is required. Thus, the values of the van der Waals surface electrostatic potential by the amide nitrogen and oxygen atoms, the atomic charge at the amide nitrogen atom, and the dipole moment must all be in optimal ranges for potent repellency. The electronic properties were calculated using the AM semi-empirical quantum chemical method using commercial software. These easily calculable predictors of repellent efficacy should be useful in predicting the relative efficacy of newly designed compounds, thus guiding the selection of new repellents for testing.

The effects of the Na(+)/Ca(++) exchange blocker on osmotic blood-brain barrier disruption.

Osmotic disruption of the blood-brain barrier (BBB) by mannitol is currently being used to enhance drug delivery in human brains. Despite clinical and experimental interest, to date the time course in the early phase of disruption has not been accurately identified. The mechanism in barrier closure also remains elusive. We first studied the rapid change in cerebrovascular permeability after BBB disruption in rats, and then demonstrated that the Na(+)/Ca(++) exchange blocker (KB-R7943) prolongs osmotic disruption. Osmotic BBB disruption was attained by using intra-arterial infusion of hypertonic mannitol in Sprague-Dawley (SD) rats. To measure the changes in cerebrovascular permeability, perfusate containing [14C]-sucrose was infused intra-arterially at different time points following osmotic stress. Cerebrovascular permeability was then measured with the in situ brain perfusion technique. This is the first in vivo study demonstrating that osmotic disruption is prolonged by the Na(+)/Ca(++) exchange blocker, which did not affect the peak level of BBB disruption. The exact time course of cerebrovascular reversibility was studied and the earliest BBB disruption was seen to occur 5 min after osmotic stress. Histopathological examination after osmotic disruption with the Na(+)/Ca(++) exchange blocker showed no neuronal damage in rat brains. Our findings represent important experimental information regarding pharmacological manipulation of BBB disruption. The possibility of prolonging the transient opening of the BBB has major clinical implications.

Quantification of early blood-brain barrier disruption by in situ brain perfusion technique.

Osmotic disruption is currently being used to circumvent the blood-brain barrier (BBB) and enhance the delivery of therapeutic molecules in human brains. To date, however, the time course during the early phase of disruption has not been clarified. In order to demonstrate the rapid change in cerebrovascular permeability after BBB disruption in rats, we developed a method of in situ brain perfusion to demonstrate the earliest reversibility in cerebrovascular permeability. Osmotic BBB disruption was attained by intracarotid infusion of hypertonic mannitol. Perfusate containing [14C]-sucrose was infused at different time points following osmotic stress followed by measuring cerebrovascular permeability. The earliest BBB disruption was seen to occur 5 min after osmotic stress, after which the exact time course of cerebrovascular reversibility was studied. The protocol reported here, in contrast with those reported in previous studies, was shown to be qualitative, simple, and fast. In addition, the method can be applied to measure any low BBB permeability molecules. This protocol should be helpful for any research focused on enhancing drug delivery into the brain following osmotic BBB disruption.

Transcranial Doppler findings during balloon test occlusion of the internal carotid artery.

The authors performed transcranial Doppler ultrasonography (TCD) during internal carotid artery (ICA) balloon test occlusion (BTO) and observed changes in mean flow velocity (Vm) in the middle cerebral artery (MCA), and pulsatility index (PI) while monitoring the stump pressure (Sp) of the internal carotid artery (ICA), and neurologic findings. A group of 17 patients requiring possible temporary or permanent occlusion of the ICA in the course of planned procedures first underwent BTO. A patient who either developed neurologic changes or maintained less than 60% of preocclusion Sp or Vm in the ipsilateral MCA during BTO was considered to have a positive test. Eleven patients had negative results, while in six patients, tests were positive. Mean flow velocity showed a decrease after occlusion in all cases but not to a remarkable extent in some patients. Stump pressure decreased in all negative cases after balloon inflation and than tended to increase progressively during 15 minutes of BTO. Pulsatility index tended to decrease gradually during BTO in all negative patients. However, in positive cases, PI and Sp fell steeply. Only one positive case had a neurologic symptom of severe headache. The decreased PI in the MCA reflected autoregulatory dilation of cerebral vessels to compensate for decreased absolute cerebral blood volume following ICA occlusion. Changes in PI are a good indicator for evaluating blood flow during BTO.

Determination of the linkages in some methylated, sialic acid-containing, meningococcal polysaccharides by mass spectrometry.

The application of gas-liquid chromatography-mass spectrometric (g.l.c.-m.s.) analysis to a number of sialic acid-containing polysaccharides of meningococcal origin has been studied. Methylation of these polysacchardies by the Hakomori conditions resulted in both O- and N-methylation. Methanolysis of the methylated polysaccharides from serogroup C [(2 leads to 9)-linked], colominic acid [(2 leads to 8)-linked], and serogroups Y and W-135 [both (1 leads to 4)-linked], yielded the respective 4,7,8-, 4,7,9-, and 7,8,9-tri-O-methyl derivatives of methyl N-acetyl-N-methyl-beta-D-neuraminate methyl glycoside. As model compounds, methyl N-acetyl-4,7,8,9-tetra-O-methyl-alpha-D-neuraminate methyl glycoside and its N-methyl derivative were also synthesized. All of the methylated derivatives could be identified on the basis of their typical fragmentation-patterns, indicating that this method is applicable to the determination of the position of linkages to sialic acid residues in biopolymers.

Structural studies on the major, capsular polysaccharide from Cryptococcus bacillisporus serotype B.

The capsular material from Cryptococcus bacillisporus serotype B has been separated into essentially two fractions. One of these (60% of the total) has been studied by the methods usual for the structural elucidation of polysaccharides. The results are consistent with a structure having nonreducing D-xylosyl as well as D-glucosyluronic acid groups attached to O-2 of D-mannosyl residues linked alpha-(1 leads to 3) in a linear backbone. Every third D-mannosyl residue is doubly substituted with a D-xylosyl group at O-2 and a D-glucosyluronic acid group at O-4.

The structure of the capsular polysaccharide from Cryptococcus neoformans serotype D.

The capsular polysaccharide from Cryptococcus neoformans serotype D has been studied by employing the usual methods for the elucidation of chemical structure. The results are consistent with the occurrence of a polysaccharide having both D-glucosyl-uronic acid and D-xylosyl groups present as nonreducing end-groups attached to O-2 of D-mannosyl residues which are linked alpha-D-(1 leads to 3) in a linear backbone.