Analysing the effect of crystal size and structure in highly efficient CH3NH3PbI3 perovskite solar cells by spatially resolved photo- and electroluminescence imaging.

CH3NH3PbI3 perovskite solar cells with a mesoporous TiO2 layer and spiro-MeOTAD as a hole transport layer (HTL) with three different CH3NH3I concentrations (0.032 M, 0.044 M and 0.063 M) were investigated. Strong variations in crystal size and morphology resulting in diversified cell efficiencies (9.2%, 16.9% and 12.3%, respectively) were observed. The physical origin of this behaviour was analysed by detailed characterization combining current-voltage curves with photo- and electroluminescence (PL and EL) imaging as well as light beam induced current measurements (LBIC). It was found that the most efficient cell shows the highest luminescence and the least efficient cell is most strongly limited by non-radiative recombination. Crystal size, morphology and distribution in the capping layer and in the porous scaffold strongly affect the non-radiative recombination. Moreover, the very non-uniform crystal structure with multiple facets, as evidenced by SEM images of the 0.032 M device, suggests the creation of a large number of grain boundaries and crystal dislocations. These defects give rise to increased trap-assisted non-radiative recombination as is confirmed by high-resolution µ-PL images. The different imaging techniques used in this study prove to be well-suited to spatially investigate and thus correlate the crystal morphology of the perovskite layer with the electrical and radiative properties of the solar cells and thus with their performance.

Structure-activity relationships of piscidin 4, a piscine antimicrobial peptide.

Piscidin 4, an antimicrobial peptide recently isolated from mast cells of hybrid striped bass (Morone chrysops female × Morone saxatilis male), is unusual in that it is twice as long (44 amino acids) as the typical members of the piscidin family. We previously showed that native piscidin 4 had a modified amino acid at position 20, but synthetic piscidin 4 (having an unmodified Trp at position 20) had similar potent activity against a number of both human and fish bacterial pathogens. In this study, the structure and membrane topology of synthetic piscidin 4 were examined using liposomes as model bilayers. Circular dichroism analyses revealed that it had a disordered structure in aqueous solution and folded to form a relatively weak α-helical structure in both membrane-mimetic trifluoroethanol solutions and liposome suspensions. Fluorescence data (piscidin 4 embedded in liposomes) and leakage experiments indicated that piscidin 4 interacted strongly with the hydrophobic part of the liposome. Binding of piscidin 4 to liposomes induced significant blue shifts of the emission spectra of the single Trp residue (Trp20). Quenching of Trp20 by water-soluble quencher (either acrylamide or I-) indicated that the fluorescence of Trp20 decreased more in the presence of liposomes than in buffer solution, thus revealing that Trp20 is less accessible to the quenchers in the presence of liposomes. The relative leakage abilities of piscidin 4 (1 µM) with liposomes were in the following order: DPPC (100%)≥EYPC (94%)>DPPC/DPPG (65%)>EYPC/EYPG (0%). This high activity against DPPC and EYPC liposomes was contrary to our data suggesting that piscidin 4 has a much weaker tendency to form an α-helix than other piscidins, such as piscidin 1. However, the structural similarity of protozoan membranes to EYPC liposomes might explain our discovery of the potent activity of piscidin 4 against the important skin/gill parasite ich (Ichthyophthirius multifiliis), but its negligible hemolytic activity against vertebrate membranes (hybrid striped bass or human erythrocytes). It also suggests that other conformation(s) in addition to the α-helix of this peptide may be responsible for its selective activity. This differential toxicity also suggests that piscidin 4 plays a significant role in the innate defense system of hybrid striped bass and may be capable of functioning extracellularly.

Structure-activity relationship of neuropeptide gamma derived from mammalian and fish.

This study of relationship between structure and biologic activity was performed using five neuropeptide gammas [NPgamma; mammalian-NPgamma (M-NPgamma), trout-NPgamma (T-NPgamma), goldfish-NPgamma (G-NPgamma), bowfin-NPgamma (B-NPgamma), and shark-NPgamma (S-NPgamma)]. Circular dichroism (CD) spectra showed that all peptides took random structure in buffer solution. In neutral and acidic liposomes, M-NPgamma, T-NPgamma, B-NPgamma, and S-NPgamma still adopted random structure, while G-NPgamma had an alpha-helical structure. The biologic activity of NPgammas has been estimated by their effects on the intestinal motility and arterial relaxation. The intestinal motility was investigated with rat duodenum (RD), carp intestine (CI), and guinea-pig ileum (GPI). The arterial relaxing effect was tested with guinea-pig aorta (GPA) and rat mesenteric artery (RMA). In RD, the order of potency compared with the EC50 value was M-NPgamma >> S-NPgamma >> B-NPgamma >> G-NPgamma >> T-NPgamma. G-NPgamma was the most contractile agent in CI. S-NPgamma was the most contractile agent in GPI. Using an arterial relaxing test, the order of potency was G-NPgamma >> T-NPgamma >> B-NPgamma >> S-NPgamma >> M-NPgamma in GPA, and all NPgammas remarkably reduced relaxing activity in RMA. Despite their structural similarities to NPgammas, G-NPgamma has high affinity to tachykinin receptor-binding sites in GPA and CI, indicating an alpha-helical structure may have a critical role for receptor binding. However, an alpha-helical structure does not play a critical role in recognizing receptor-binding sites in RD and GPI.

Morphological and photoelectrochemical characterization of core-shell nanoparticle films for dye-sensitized solar cells: Zn-O type shell on SnO2 and TiO2 cores.

Core-shell type nanoparticles with SnO2 and TiO2 cores and zinc oxide shells were prepared and characterized by surface sensitive techniques. The influence of the structure of the ZnO shell and the morphology ofnanoparticle films on the performance was evaluated. X-ray absorption near-edge structure and extended X-ray absorption fine structure studies show the presence of thin ZnO-like shells around the nanoparticles at low Zn levels. In the case of SnO2 cores, ZnO nanocrystals are formed at high Zn/Sn ratios (ca. 0.5). Scanning electron microscopy studies show that Zn modification of SnO2 nanoparticles changes the film morphology from a compact mesoporous structure to a less dense macroporous structure. In contrast, Zn modification of TiO2 nanoparticles has no apparent influence on film morphology. For SnO2 cores, adding ZnO improves the solar cell efficiency by increasing light scattering and dye uptake and decreasing recombination. In contrast, adding a ZnO shell to the TiO2 core decreases the cell efficiency, largely owing to a loss of photocurrent resulting from slow electron transport associated with the buildup of the ZnO surface layer.

Structures of neuropeptide gamma from goldfish and mammalian neuropeptide gamma, as determined by 1H NMR spectroscopy.

Neuropeptide gamma belongs to tachykinin families which have a common C-terminal amino acid sequence (Phe-X-Leu-Met-NH2) and which induce various biological responses including salivation, hypotension, and contraction of gastrointestinal, respiratory, and urinary smooth muscle. In the present study, we present the solution structures of neuropeptide gamma (NPgamma) from gold fish (G-NPgamma) and mammalian NPgamma (M-NPgamma), as determined by nuclear magnetic resonance (NMR) spectroscopy in 50% trifluoroethanol (TFE)/water (1 : 1, v/v) solution and 200 mm sodium dodecyl sulfate (SDS) micelles. In aqueous TFE solution, G-NPgamma has a alpha-helical conformation in the region of His12-Met21 and a short helix in the N-terminal region, and has a beta-turn from Arg9 to Arg11 in between. In aqueous TFE solution, M-NPgamma also has alpha-helical conformations both in the C-terminal region and the N-terminal region and a beta-turn from His9 to Arg11 in between. In SDS micelle, the structure of G-NPgamma contains a stable alpha-helix from His12 to Met21 and a beta-turn from Arg9 to Arg11, while M-NPgamma has a short helix from Ser16 to Met21. The region from His12 to Met21 corresponds to the amino acid sequence of neurokinin A. Neuropeptide gamma may act as a precursor of neurokinin A and the post-translational processing of this peptide involves the enzymatic attack of the basic beta-turn region from residue 9 to residue 11 in the middle. From our relaxation study, it could be suggested that in fish system G-NPgamma induces the biological actions corresponding to those of substance P in mammalian system. The structures of G-NPgamma and M-NPgamma contain alpha-helical structures at the C-terminus and this helix seems to promote the affinity for NK1 and/or NK2 receptor.

Effects of mastoparan B and its analogs on the phospholipase D activity in L1210 cells.

Mastoparan B (MP-B), an amphiphilic alpha-helical peptide isolated from hornet venom, and its Ala-substituted analogs were examined for their effectiveness on phospholipase D (PLD) activity in L1210 cells. PLD activity was determined by measuring phosphatidylethanol produced from [3H]myristate-labelled cells in the presence of ethanol. PLD activity was stimulated by MP-B, 4MP-B (Lys4-->Ala), and 12MP-B (Lys12-->Ala), but not by 3MP-B (Leu3-->Ala) and 9MP-B (Trp9-->Ala). Other MPs including mastoparan 7 also stimulated the PLD activity, but inactive mastoparan 17 did not. The stimulatory effect of various MP analogs could be correlated with their alpha-helical contents. The PLD activity stimulated by MP-B was not affected by G-protein blocking chemicals. The extent of PLD stimulation by various MP-Bs, as well as by digitonin and beta-escin, correlated with the permeability of the membrane to ethidium bromide. These results suggest that the stimulation of PLD activity by MP-B in L1210 cells is probably coupled with membrane perturbation brought about by the peptide.

Interaction of mastoparan-B from venom of a hornet in Taiwan with phospholipid bilayers and its antimicrobial activity.

Mastoparan B (MP-B), an amphiphilic alpha-helical peptide newly isolated from the hornet Vespa basalis, was studied in comparison with mastoparan (MP), in terms of interaction with the phospholipid bilayer and of hemolytic and antimicrobial activity. The amphiphilic structure of MP-B has more hydrophilic amino acid residues in the hydrophilic surface than that of MP. Although each peptide had a considerably different effect on the interaction with lipid bilayers (e.g., their conformation in the presence of acidic and of neutral lipids and dye-release ability from the encapsulated liposomes), on the whole the interaction mode was similar. MP-B caused a change in the shape of erythrocytes from normal discoid to a crenated form (named echinocytes). MP exhibited strong activity against gram-positive bacteria but not against gram-negative ones. Contrary to this, MP-B showed both strong activity against gram-positive bacteria and potent activity against gram-negative bacteria. Whereas both peptides have almost the same residues on the hydrophobic side, the difference in the hydrophilic surface area on the molecules seems to lead to the subtle change in its interaction with membranes, resulting in the alteration of biological activity.

Role of basic residues for the binding of omega-conotoxin GVIA to N-type calcium channels.

Each of four basic residues of omega-conotoxin GVIA was replaced with alanine to study the role of basic residues for the binding of this toxin to N-type calcium channels. The activities of these analogs were estimated from the inhibitory action on 125I-omega-conotoxin GVIA binding to chick brain synaptic plasma membranes. The replacement of Arg17, Lys24 and Arg25 resulted in no significant change in the activity and all of the analogs gave the same IC50 value (0.15 nM) as that of native omega-conotoxin GVIA. The inhibitory action of [Ala2]omega-conotoxin GVIA (K2A) was 40-times less potent (IC50 = 5.5 nM); however, full inhibition was achieved at a concentration above 0.1 microM. These results indicate that the Arg residue is not essential for the activity of omega-conotoxin GVIA. The nature of association to ion channels may be different between omega-conotoxin GVIA and mu-conotoxin GIIIA.

Conformation of tachyplesin I from Tachypleus tridentatus when interacting with lipid matrices.

The mode of action of tachyplesin I, an antimicrobial cationic heptadecapeptide amide isolated from the hemocyte debris of a horseshoe crab, Tachypleus tridentatus, toward lipid matrices was studied with synthetic tachyplesin I, its analogs with Phe in place of Trp or Tyr, a linear analog with no disulfide bonds, and two linear short fragments. Circular dichroism spectra showed that tachyplesin I took an antiparallel beta-structure in buffer solution and a certain less ordered structure in acidic liposomes composed of egg phosphatidylcholine and egg phosphatidylglycerol (3:1). Spectrophotometric titration of the peptides with laurylphosphorylcholine revealed that both Trp and Tyr residues orient toward the inside of lipid matrices, suggesting that they are on the same side of the peptide backbone. The carboxyfluorescein leakage experiment and fluorescence data indicated that tachyplesin I interacted strongly with neutral and acidic lipid bilayers and an aromaticity-rich hydrophobic part of the peptide was embedded in lipid membranes. All the peptides except for the short fragments were almost equally active in lipopolysaccharide binding. The energy-transfer experiment showed that a conformational change occurred such that the Tyr and Trp residues are positioned more closely to each other in acidic liposomes than in buffer solution. The present study strongly suggested that amphipathic lipid bilayers induced a conformational change of tachyplesin I from an energetically stable beta-structure to a less ordered, probably more amphipathic structure.

Basic amphipathic helical peptides induce destabilization and fusion of acidic and neutral liposomes.

We have studied the fusion of small unilamellar vesicles composed of egg PC and of a mixture of egg PC plus egg PA using various basic amphipathic peptides. Fusion was monitored by carboxyfluorescein leakage assay, light scattering, membrane intermixing assay, contents mixing assay and electron microscopy. Ac-(L-Leu-L-Ala-L-Arg-L-Leu)3-NHCH3 (peptide 4(3] and Ac-(L-Leu-L-Ala-L-Lys-L-Leu)3-NHCH3 (peptide 4'3), which have high hydrophobic moments, caused transformation of small unilamellar vesicles into larger and relatively homogeneous ones. Ac-(L-Leu-L-Leu-L-Ala-L-Arg-L-Leu)2-NHCH3 (5(2], which has medium hydrophobic moment, induced weak but appreciable fusion, while Ac-(L-Ala-L-Arg-L-Leu)3-NHCH3 (3(3] which has no helical structure did not show any fusion. However, peptides 4(3), 4'3 and 5(2) caused massive leakage of the contents from small unilamellar vesicles. These results indicated that interaction of the peptides with artificial membranes caused extensive perturbation of the lipid bilayer, followed by fusion. The fusogenic capacity of model basic peptides was correlated with the hydrophobic moment of each peptide when the peptides adopted an alpha-helical structure in the presence of acidic liposomes. Peptides 4(3) and 4'3 also showed weak fusogenic ability for neutral liposomes, while 5(2) and 3(3) showed no ability, suggesting that highly amphipathic peptides, such as 4(3), interact weakly but distinctly with neutral liposomes to fuse them.

Effects of model extension peptides containing glycine, proline and serine residues on the import of mitochondrial enzyme precursors into mitochondria.

We found previously that Ac-(Leu-Leu-Ala-Arg-Leu)3-NHCH3 (5(3)), a model of the extension peptide of cytochrome P-450(SCC) precursor, strongly inhibited the import of the precursor into mitochondria. Unexpectedly, however, 5(3) showed the break of respiratory control of mitochondria. In the present study, desAc- [Pro7, Ser10] 5(3) (1), [Pro7, Ser10] 5(3) (2), desAc- [Gly5, Pro7, Ser10] 5(3) (3) and [Gly5, Pro7, Ser10] 5(3) (4) were synthesized in order to investigate the influence of the Gly, Pro and Ser residues, which are present in the extension peptide, on the import and respiration. CD measurement indicated that all the peptides had an alpha-helical conformation in the presence of DPPC-DPPG (3:1) liposomes. The order of the content of alpha-helical conformation was 5(3) greater than 1, 2 greater than 3, 4, while SEP1-15 (N-terminal 1-15 fragment of the extension peptide of cytochrome P-450(SCC) precursor) showed no alpha-helical structure. The measurements of dye-leakage from liposomes, import-inhibition of P-450(SCC) and adrenodoxin precursors and respiratory inhibition of mitochondria indicated that 3 and 4, rather than 1 and 2, are similar to SEP1-15, and that the glycine residue in the extension peptide is of considerable importance for the import of the precursors.

Interaction of basic extension peptide fragments of adrenodoxin precursor with phospholipid vesicles.

Two extension peptide fragments PA1-4 and PA17-32, which correspond to the residues 1-14 and 17-32, respectively, of adrenodoxin precursor, were synthesized by the solution method to find a sequence necessary for the import of the precursor into mitochondria. Biological assay showed that PA1-14 inhibited the import of two mitochondrial enzyme precursors, but PA17-32 showed no inhibition, indicating that the N-terminal sequence has important information for import. CD spectra of the peptides demonstrated that PA1-14 formed alpha-helical structure in Tris-HCl buffer (pH 7.4) containing acidic phospholipid liposomes. Furthermore, PA1-14 induced the moderate leakage of carboxyfluorescein from phospholipid vesicles. The relationship between the structure and function of the peptides is discussed.