E. J. Padma Malar

Synthesis, pharmacological screening, quantum chemical and in vitro permeability studies of N-Mannich bases of benzimidazoles through bovine cornea.

A novel series of N-Mannich bases of benzimidazole derivatives were synthesized and characterized by (1)H NMR, IR spectral studies and elemental analysis. The compounds were screened for analgesic and anti-inflammatory activity. 1-((Diethylamino)-methyl)-2-styryl benzimidazole 4 at 40mg/kg was found to be equipotent to paracetamol. 1-((Piperidin-1-yl) methyl)-2-styryl-benzimidazole 6 at 40mg/kg was found to be more potent than Diclofenac. Corneal permeability and quantum chemical calculations were performed to correlate the hydrogen bonding ability with permeability and activity. The energies of the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) were correlated with pharmacological activity. The semi-empirical PM3 calculations (quantum chemical calculations) revealed that E(LUMO) and energy gap DeltaE were capable of accounting for the high in vitro bovine corneal permeability and activity of the compounds.

Excited states of antiaromatic systems1

Abstract SINDO1 studies were performed to optimize the geometry of excited states of some antiaromatic molecules. It is discussed how such states can exhibit aromatic character upon suitable electronic excitation. The nodal pattern of the molecular orbitals involved in the electronic excitation are used to invoke bond equilization in excited states. We have investigated singlet and triplet excited states of five-membered rings C 4 H 5 B, C 5 H 5 + and C 5 H 4 O containing four π electrons and bicyclic systems bicyclo-(1,1,0)-butadiene, bicyclo-(2,2,0)-hexatriene and benzocylobutadiene. It is seen that in bicyclo-(2,2,0)-hexatriene, both the bicylic structure and the 1,4-diradical structure determine the equilibrium geometry.

Lipid-induced conformational transition of the amyloid core fragment Aβ(28–35) and its A30G and A30I mutants

The interaction of the β‐amyloid peptide (Aβ) with neuronal membranes could play a key role in the pathogenesis of Alzheimer’s disease. Recent studies have focused on the interactions of Aβ oligomers to explain the neuronal toxicity accompanying Alzheimer’s disease. In our study, we have investigated the role of lipid interactions with soluble Aβ(28–35) (wild‐type) and its mutants A30G and A30I in their aggregation and conformational preferences. CD and Trp fluorescence spectroscopic studies indicated that, immediately on dissolution, these peptides adopted a random coil structure. Upon addition of negatively charged 1,2‐dipalmitoyl‐syn‐glycero‐3‐phospho‐rac‐(glycerol) sodium salt (PG) lipid, the wild‐type and A30I mutant underwent reorganization into a predominant β‐sheet structure. However, no conformational changes were observed in the A30G mutant on interaction with PG. In contrast, the presence of zwitterionic 1,2‐dipalmitoyl‐syn‐glycero‐3‐phosphatidylcholine (PC) lipid had no effect on the conformation of these three peptides. These observations were also confirmed with atomic force microscopy and the thioflavin‐T assay. In the presence of PG vesicles, both the wild‐type and A30I mutant formed fibrillar structures within 2 days of incubation in NaCl/Pi, but not in their absence. Again, no oligomerization was observed with PC vesicles. The Trp studies also revealed that both ends of the three peptides are not buried deep in the vesicle membrane. Furthermore, fluorescence spectroscopy using the environment‐sensitive probe 1,6‐diphenyl‐1,3,5‐hexatriene showed an increase in the membrane fluidity upon exposure of the vesicles to the peptides. The latter effect may result from the lipid head group interactions with the peptides. Fluorescence resonance energy transfer experiments revealed that these peptides undergo a random coil‐to‐sheet conversion in solution on aging and that this process is accelerated by negatively charged lipid vesicles. These results indicate that aggregation depends on hydrophobicity and propensity to form β‐sheets of the amyloid peptide, and thus offer new insights into the mechanism of amyloid neurodegenerative disease.

Photophysical Studies on the Interaction of Acridinedione Dyes with Universal Protein Denaturant: Guanidine Hydrochloride

Photophysical studies of photoinduced electron transfer (PET) and non-PET based acridinedione dyes with guanidine hydrochloride (GuHCl) were carried out in water and methanol. Addition of GuHCl to photoinduced electron transfer (PET) based acridinedione dye (ADR 1) results in a fluorescence enhancement, whereas a non-PET based dye (ADR 2) shows no significant change in the fluorescence intensity and lifetime. Addition of GuHCl to ADR 1 dye in methanol results in single exponential decay behaviour, on the contrary a biexponential decay pattern was observed on the addition of GuHCl in water. Absorption and emission spectral studies of ADR 1 dye interaction with GuHCl reveals that the dye molecule is not in the protonated form in aqueous GuHCl solution, and the dye is confined to two distinguishable microenvironment in the aqueous phase. A large variation in the microenvironment around the dye molecule is created on the addition of GuHCl and this was ascertained by time-resolved area normalized emission spectroscopy (TRANES) and time-resolved emission spectroscopy (TRES). The dye molecule prefers to reside in the hydrophobic microenvironment, rather in the hydrophilic aqueous phase is well emphasized by time-resolved fluorescence lifetime studies. The mechanism of fluorescence enhancement of ADR 1 dye by GuHCl is attributed to the suppression of the PET process occuring through space.

Lipid-Induced Conformational Transition of Amyloid β Peptide Fragments

Conformational transition of soluble monomeric amyloid β-peptide (Aβ) into oligomeric and protofibrillar aggregates plays a key role in the pathogenesis of Alzheimers disease (AD). One of the central questions surrounding the molecular pathophysiology of AD is how the soluble Aβ is converted into its aggregated toxic form. A more detailed understanding of the conformational transitions involved in the self-assembly of Aβ may facilitate the design of inhibitors of aggregation. In this study, we evaluated the wild-type (WT) Aβ 16–28 peptide (KLVFFAEDVGSNK) and its associated mutants, including A21G (Flemish), E22K (Italian), E22Q (Dutch), and E22G (Arctic) mutants, by examining, in particular, their aggregation kinetics in the presence and in the absence of negatively charged and zwitterionic lipids. Circular dichroic and thioflavin T fluorescence studies indicated that the WT peptide undergoes a rapid conformational transition into β-sheet structure in solution, whereas the Arctic and Dutch variants show a markedly rapid transition into β-sheet structure in the presence of negatively charged lipids. These results provide strong evidence suggesting that the reduction in net charge, with a concurrent increase in the net hydrophobicity of the peptide alone or when complexed with lipid in solution, determines the rate of aggregate formation.

Preparation, physiochemical characterization, and oral immunogenicity of Aβ(1–12), Aβ(29–40), and Aβ(1–42) loaded PLG microparticles formulations

Alzheimers disease (AD) is caused by the deposition of beta-amyloid (Abeta) protein in brain. The current AD immunotherapy aims to prevent Abeta plaque deposition and enhance its degradation in the brain. In this work, the peptides B-cell epitope Abeta(1-12), T-cell epitope Abeta(29-40) and full-length Abeta(1-42) were loaded separately to the poly (D,L-lactide co-glycolide) (PLG) microparticles by using W/O/W double emulsion solvent evaporation method with entrapment efficacy of 70.46%, 60.93%, and 65.98%, respectively. The prepared Abeta PLG microparticles were smooth, spherical, individual, and nonporous in nature with diameters ranging from 2 to 12 microm. The cumulative in vitro release profiles of Abeta(1-12), Abeta(29-40), and Abeta(1-42) from PLG microparticles sustained for long periods and progressively reached to 73.89%, 69.29%, and 70.08% by week 15. In vitro degradation studies showed that the PLG microparticles maintained the surface integrity up to week 8 and eroded completely by week 16. Oral immunization of Abeta peptides loaded microparticles in mice elicited stronger immune response by inducing anti-Abeta antibodies for prolonged time (24 weeks). The physicochemical characterization and immunogenic potency of Abeta peptides incorporated PLG microparticles suggest that the microparticles formulation of Abeta can be a potential oral AD vaccine.

Aggregation and conformational studies on a pentapeptide derivative

Most of the disease causing proteins such as beta amyloid, amylin, and huntingtin protein, which are natively disordered, readily form fibrils consisting of beta-sheet polymers. Though all amyloid fibrils are made up of beta-sheet polymers, not all peptides with predominant beta-sheet content in the native state develop into amyloid fibrils. We hypothesize that stable amyloid like fibril formation may require mixture of different conformational states in the peptide. We have tested this hypothesis on amyloid forming peptide namely HCl(Ile)(5)NH(CH(2)CH(2)O)(3)CH(3) (I). We show peptide I, has propensity to form self-assembled structures of beta-sheets in aqueous solutions. When incubated over a period of time in aqueous buffer, I self assembled into beta sheet like structures with diameters ranging from 30 to 60 A that bind with amyloidophilic dyes like Congo red and Thioflavin T. Interestingly peptide I developed into unstable fibrils after prolonged aging at higher concentration in contrast with the general mature fibril-forming propensity of various amyloid petides known to date.

Investigation of aromaticity in the excited states of fulvene

Abstract The low-lying excited states of fulvene are examined by configuration interaction (CI) calculations using the SINDO1 method. The structural optimization reveals major reorganization in the ring geometry in the excited states. The lowest excited singlet ( 1 B 2 ) and triplet ( 3 B 2 ) states are found to be moderately aromatic with a substantial contribution from the dipolar aromatic structure.

Research ArticlesPreparation, physiochemical characterization, and oral immunogenicity of Aβ(1–12), Aβ(29–40), and Aβ(1–42) loaded PLG microparticles formulations

Alzheimers disease (AD) is caused by the deposition of beta-amyloid (Abeta) protein in brain. The current AD immunotherapy aims to prevent Abeta plaque deposition and enhance its degradation in the brain. In this work, the peptides B-cell epitope Abeta(1-12), T-cell epitope Abeta(29-40) and full-length Abeta(1-42) were loaded separately to the poly (D,L-lactide co-glycolide) (PLG) microparticles by using W/O/W double emulsion solvent evaporation method with entrapment efficacy of 70.46%, 60.93%, and 65.98%, respectively. The prepared Abeta PLG microparticles were smooth, spherical, individual, and nonporous in nature with diameters ranging from 2 to 12 microm. The cumulative in vitro release profiles of Abeta(1-12), Abeta(29-40), and Abeta(1-42) from PLG microparticles sustained for long periods and progressively reached to 73.89%, 69.29%, and 70.08% by week 15. In vitro degradation studies showed that the PLG microparticles maintained the surface integrity up to week 8 and eroded completely by week 16. Oral immunization of Abeta peptides loaded microparticles in mice elicited stronger immune response by inducing anti-Abeta antibodies for prolonged time (24 weeks). The physicochemical characterization and immunogenic potency of Abeta peptides incorporated PLG microparticles suggest that the microparticles formulation of Abeta can be a potential oral AD vaccine.

Kinetic measurements and quantum chemical calculations on low spin Ni(II)/(III) macrocyclic complexes in aqueous and sulphato medium

AbstractCu(II) ion catalyzed kinetics of oxidation of H2O2 by [NiIIIL2