Circular dichroism and magnetic circular dichroism spectra of chlorophylls in nematic liquid crystals. I. Electric and weak magnetic field effects on the dichroism spectra

Dichroism spectra of chlorophyll a, chlorophyll b and bacteriochlorophyll a in various nematic liquid crystals are reported. The initial orientation of chlorophylls in such a sample is determined by the interaction of the aggregate formed from the pigment and the liquid crystal molecules with the electrode surface on the cell windows. Reorientation is carried out by either an electric or magnetic field. The analysis of the circular dichroism spectra obtained from these samples on the basis of the Mueller matrix shows that the intensity is predominantly related to the texture of the sample. Chlorophyll molecules can be aggregated with liquid crystals in two ways: (1) through the chlorin magnesium atom, which results in the liquid crystal chain being almost perpendicular to the porphyrin ring, or (2) attached parallel to the line connecting the first and third pyrrole rings of the chlorin, the chlorin now lying in the plane of the liquid crystal chains. By comparing the dichroism spectra of various chlorophylls in the same liquid crystal we can draw conclusions concerning the preferred type of aggregation, not only with liquid crystals, but also with biological molecules. These liquid crystal systems are models of the orientation effects found for chlorophyll in lamellae. The model studied in this work is much simpler than the lamellar system but it does exhibit several common properties with the latter. Both systems are anisotropic and show much more intense dichroism signals, often of opposite sign, compared with those observed for photosynthetic pigments in isotropic solutions. Dichroism signals of organism fragments are much more complex than those of our model, which can either be related to the occurrence in the organism of several types of pigments or, for a given type of pigment, could be the result of exciton splitting. On the basis of our model it is shown that small changes in the anisotropy of the pigment in the surroundings have a strong influence on the sign and amplitude of the observed circular dichroism signal. Such effects may be responsible for the structure of the dichroism spectra observed for biological samples. Such structures can be partially related to the superposition of the dichroism signal from various ‘domains’ of chromophore which are different in both pigment arrangement and in the anisotropy of the surroundings of the pigment molecules themselves.     

Analysis of Skeletal Muscle Defects in Larval Zebrafish by Birefringence and Touch-evoke Escape Response Assays

Zebrafish (Danio rerio) have become a particularly effective tool for modeling human diseases affecting skeletal muscle, including muscular dystrophies^1-3, congenital myopathies^4,5, and disruptions in sarcomeric assembly^6,7, due to high genomic and structural conservation with mammals⁸. Muscular disorganization and locomotive impairment can be quickly assessed in the zebrafish over the first few days post-fertilization. Two assays to help characterize skeletal muscle defects in zebrafish are birefringence (structural) and touch-evoked escape response (behavioral).Birefringence is a physical property in which light is rotated as it passes through ordered matter, such as the pseudo-crystalline array of muscle sarcomeres⁹. It is a simple, noninvasive approach to assess muscle integrity in translucent zebrafish larvae early in development. Wild-type zebrafish with highly organized skeletal muscle appear very bright amidst a dark background when visualized between two polarized light filters, whereas muscle mutants have birefringence patterns specific to the primary muscular disorder they model. Zebrafish modeling muscular dystrophies, diseases characterized by myofiber degeneration followed by repeated rounds of regeneration, exhibit degenerative dark patches in skeletal muscle under polarized light. Nondystrophic myopathies are not associated with necrosis or regenerative changes, but result in disorganized myofibers and skeletal muscle weakness. Myopathic zebrafish typically show an overall reduction in birefringence, reflecting the disorganization of sarcomeres.The touch-evoked escape assay involves observing an embryo''s swimming behavior in response to tactile stimulation^10-12. In comparison to wild-type larvae, mutant larvae frequently display a weak escape contraction, followed by slow swimming or other type of impaired motion that fails to propel the larvae more than a short distance¹². The advantage of these assays is that disease progression in the same fish type can be monitored in vivo for several days, and that large numbers of fish can be analyzed in a short time relative to higher vertebrates.     

传统光学相干断层成像:软骨双折射特性的研究

传统光学相干断层成像可实现无损伤在体检测,具有较高的分辨率和灵敏度。获取生物组织的双折射信息可望是其应用之一。本文用传统光学相干断层成像系统以动物模型对关节软骨进行了研究,对软骨组织的传统光学相干断层图像的层结构随脱水和机械作用的变化进行了分析。结果表明,传统光学相干断层成像系统在一定程度上可用于监测由其胶原质纤维排列决定的软骨组织的双折射和密度。     

利用偏光影像技术定量观察大鼠胸主动脉中纤维结构

目的：本实验选取大鼠胸主动脉为研究对象,探讨定量偏光影像技术对双折光性物质的成像特点与适用研究领域。方法：采用Abrio液晶偏光影像系统在未经染色条件下对青年和衰老大鼠胸主动脉纤维结构进行观察,然后对主动脉切片进行苦味酸天狼猩红染色,对比Abrio和传统偏振光技术成像的差别。结果：较传统偏振光显微镜不同,Abfio可以对纤维双折射的方位角和光程差进行定量分析。在未经染色的条件下,Abrio可对大鼠胸主动脉中具有双折光性的纤维结构清晰成像,而传统偏光显微镜在未经染色条件下不能给出有意义的信息。经苦味酸天狼猩红染色后,对比传统偏振光技术,Abrio使成像不受纤维排列方向和人为因素的影响,较为完整地反映主动脉中纤维的结构及分布情况。结论：Abrio液晶偏光影像技术能够实现对样本双折光的强弱及方向的定性和定量分析,在未经染色的条件下给出微观结构信息,适合于心血管疾病的检测与评估,可拓展应用面宽,将在未来生物医药领域中体现更大的价值。     

Flexibility of myosin in pyrophosphate and NaCl solutions

The orientational relaxation time of myosin has been reported as 38 s when measured in pyrophosphate media at elevated pH (Hvidt et al. 1984) and as 17 s when measured in 0.3 M KCl at pH 7.3 (Bernengo and Cardinaud 1982). This discrepancy, which is reexamined in the present report, suggests that in KCl solution the rod portion of the myosin molecule is bent with an average angle close to 110°, whereas in pyrophosphate at elevated pH it assumes a nearly straight and rigid conformation. Electric birefringence shows that the amount of dimeric and polymeric species in pyrophosphate media at pH's 8.0 and 8.5 is certainly greater than usually thought. In these media, relaxation times can be measured correctly at pH 9.0. A comparative analysis of the influence of protein concentration, field strength, medium composition and concentration, pH and temperature showed that a high relaxation time is associated with the presence of pyrophosphate and that the myosin tail is significantly stiffened in the presence of this anion. Offprint requests to: R. Cardinaud     

Mechanism of chlorpromazine binding by Gram-positive and Gram-negative bacteria

Chlorpromazine forms charge-transfer complexes with xanthene dyes in bacteria. These complexes permit the differentiation of Gram-positive and Gram-negative bacteria in both light and polarization microscopy. The birefringence induced by the charge-transfer complex might explain the molecular basis of bacterial staining.The charge-transfer complexes formed between chorpromazine and xanthene dyes accumulate in the bacterial cell, mainly inside the bacterial cell wall. The complexes give the cells a color, which depends on the chemical composition of the staining structure, and in particular the polysaccharides of the cell wall in bacteria.Metachromatic granules were seen inside Gram-positive bacteria after chlorpromazine and rose bengal staining. Although the nature of these granules remains unclear, this type of binding may have a role in the inhibition of biochemical processes in the bacterial cells.     

Thioflavin T fluorescence assay for β-lactoglobulin fibrils hindered by DAPH

The molecule 4,5-dianilinophthalimide was recently found to be an efficient compound in disaggregating amyloid fibrils involved in the Alzheimer’s disease. In this study we have investigated whether the compound 4,5-dianilinophthalimide was able to disaggregate fibrils derived from β-lactoglobulin. In addition to a Thioflavin T fluorescence assay, flow-induced birefringence was used as an independent technique to measure the total length concentration of the fibrils. An additional advantage of the latter technique is that not only the total length concentration, but also the length distribution of the fibrils can be measured. The results from flow-induced birefringence showed that the total amount of fibrils and also the length distribution of the fibrils was not influenced by the addition of 4,5-dianilinophthalimide, even though this was suggested by the results of the Thioflavin T assay. The results of flow-induced birefringence were confirmed by rheological measurements and transmission electron microscopy. Our findings show that the use of a Thioflavin T assay in order to probe the possible disaggregating effect of certain compounds can give misleading results.     

l-Ala Modified Analogues of Amyloid β-Peptide Residue 17-20: Self-Association and Amyloid-like Fibril Formation

l-Ala modified analogues of amyloid β-peptide residue 17-20 LVFF (-l-Leu-l-Val-l-Phe-l-Phe-) have been designed and synthesized to study their self-assembling propensity, the nature of intermolecular interactions and rationalize with short hydrophobic sequences in the middle of Aβ that have important role in the neuropathology of Alzheimer’s disease. The peptides sequences LVFA and LAFA have been adopted from the β-sheet region of non-amyloidogenic proteins (hemoglobin-like falvoprotein and ATP synthase C chain, respectively). All the reported peptides self-associate into amyloid-like fibrils which are readily stained with a physiological dye Congo red and exhibits green gold birefringence under polarized light. The solid state FTIR studies of the fibrils reveal that the reported peptides self-associate through intermolecular hydrogen bonds to form antiparallel β-sheet structure, which is also supported by molecular modeling studies. This result suggests that l-Ala analogous of Aβ17-20, LVFA and LAFA also have virtually identical aggregation behavior.     

Towards the Accurate and Efficient Calculation of Optical Rotatory Dispersion Using Augmented Minimal Basis Sets

It has been recognized that quantum‐chemical predictions of dispersive (nonresonant) chiroptical phenomena are exquisitely sensitive to the periphery of the electronic wavefunction. To further elaborate and potentially exploit this assertion, linear‐response calculations of specific optical rotation were performed within the framework of density‐functional theory (DFT) by augmenting small basis sets (e.g., STO ‐ 3G and 3 ‐ 21G) for the core and valence electrons with diffuse functions taken from substantially larger bases (e.g., aug‐cc‐pVXZ where X = D, T, or Q). Of particular interest was the ability of such computationally efficient (augmented small‐basis) model chemistries to reproduce results derived from more expensive (canonical large‐basis) schemes. The results appear to be quite promising, with the augmented minimal‐basis ansatz often yielding wavelength‐resolved rotatory powers close to those deduced from standard DFT(B3LYP)/aug‐cc‐pVXZ treatments. Analogous linear‐response analyses were performed by means of coupled‐cluster singles and doubles (CCSD) theory, once again leading to augmented small‐basis estimates of specific rotation in reasonable accord with their large‐basis counterparts. Although CCSD predictions were deemed to be slightly worse than those obtained from DFT, they still were of sufficient quality for such reduced‐basis calculations to be considered viable for exploratory work. Chirality 25:606–616, 2013. © 2013 Wiley Periodicals, Inc.     

Polarization sensitivity as a contrast enhancer in pelagic predators: lessons from in situ polarization imaging of transparent zooplankton

Because light in the pelagic environment is partially polarized, it has been suggested that the polarization sensitivity found in certain pelagic species may serve to enhance the contrast of their transparent zooplankton prey. We examined its potential during cruises in the Gulf of Mexico and Atlantic Ocean and at a field station on the Great Barrier Reef. First, we collected various species of transparent zooplankton and micronekton and photographed them between crossed polarizers. Many groups, particularly the cephalopods, pelagic snails, salps and ctenophores, were found to have ciliary, muscular or connective tissues with striking birefringence. In situ polarization imagery of the same species showed that, while the degree of underwater polarization was fairly high (approx. 30% in horizontal lines of sight), tissue birefringence played little to no role in increasing visibility. This is most likely due to the low radiance of the horizontal background light when compared with the downwelling irradiance. In fact, the dominant radiance and polarization contrasts are due to unpolarized downwelling light that has been scattered from the animal viewed against the darker and polarized horizontal background light. We show that relatively simple algorithms can use this negative polarization contrast to increase visibility substantially.