Orientation and spectral properties of two stilbazolium merocyanine dyes in stretched and unstretched polyvinyl alcohol films

Spectral properties (anisotropy coefficients calculated for absorption, emission and fluorescence decay time) of two stilbazolium merocyanine dyes have been determined to evaluate the applicability of these dyes as sensitizers in photodynamic therapy. The dyes were embedded in an anisotropic polymer matrix. Analysis of the emission decay components measured in polarized light provides information on the interactions of the dye molecules with the polymer matrix being a model of an anisotropic biological system. Different values of the emission anisotropies obtained from various polarized components of fluorescence decays have shown that the orientations of the dye molecules influence their interactions with the polymer. This means that differently oriented dye molecules located in biological systems should exhibit different interactions with membranes. The chain length and type of side groups attached as well as the salt form of the dye molecule were shown to influence the dye-polymer interactions and should be taken into account before the application of merocyanine dyes in medicine. These dyes seem to be promising optical sensors with spectral properties, including the calculated anisotropy coefficients, sensitive to the molecular environment, useful to study orientation and interaction with neighbouring molecules in biological membranes.     

Correlation between structural and functional changes in brain in an idiopathic headache syndrome.

Fundamental to the concept of idiopathic or primary headache, including migraine, tension-type headache and cluster headache, is the currently accepted view that these conditions are due to abnormal brain function with completely normal brain structure. Cluster headache is one such idiopathic headache with many similarities to migraine, including normal brain structure on magnetic resonance imaging and abnormal function in the hypothalamic grey matter by positron emission tomography. Given the consistency of the positron emission tomography findings with the clinical presentation, we sought to assess whether the brains of such patients were structurally normal. We used voxel-based morphometry, an objective and automated method of analyzing changes in brain structure, to study the structure of the brains of patients with cluster headache. We found a co-localization of structural changes and changes in local brain activity with positron emission tomography in the same area of the brain in the same patients. The results indicate that the current view of the neurobiology of cluster headache requires complete revision and that this periodic headache is associated with a hitherto unrecognized brain abnormality in the hypothalamic region. We believe that voxel-based morphometry has the potential to change in the most fundamental way our concept of primary headache disorders, requiring a radical reappraisal of the tenet of structural normality.     

Cortical and subcortical localization of response to pain in man using positron emission tomography.

A quantitative study of the regional cerebral responses to non-painful and painful thermal stimuli in six normal volunteers has been done by monitoring serial measurements of regional blood flow measured by positron emission tomography (PET). In comparison to a baseline of warm stimulation no statistically significant changes in blood flow were seen in relation to increasing non-painful heat. However, highly significant increases in blood flow were seen in response to painful heat in comparison to non-painful heat. These changes were in the contralateral cingulate cortex, thalamus and lenticular nucleus. These findings are discussed in relation to previous physiological observations of responses to nociceptive stimuli in man and primates.     

In Vivo Binding to Peripheral Benzodiazepine Binding Sites in Lesioned Rat Brain: Comparison Between [3H]PK11195 and [18F]PK14105 as Markers for Neuronal Damage

Peripheral-type benzodiazepine binding sites are not normally present in most cerebral tissues, but following neuronal damage, the cells involved in the ensuing gliosis show a marked expression of these sites. In a unilateral excitotoxic striatal lesion in the rat, we sought to determine whether the isoquinoline derivatives PK11195 and PK14105 bind to these sites in vivo and whether demonstration of these sites offers the potential of indirectly localising areas of neuronal damage. Binding was studied at several intervals after coinjection of [3H]PK11195 and [18F]PK14105 to determine the time courses of specific binding. Both compounds were rapidly extracted into all cerebral tissues, but in the absence of binding sites in nonlesioned tissues, this was followed by a rapid clearance of radioactivity. In lesioned areas, both [3H]PK11195 and [18F]PK14105 accumulated over the first 5 min followed by a much slower clearance of radioactivity, resulting in a "specific signal." [3H]PK11195 binding peaked at 20-30 min postinjection, with radioactivity in the lesioned striatum being three times greater than in its contralateral homologue. The specific signal was present for at least 60 min. The maximal [18 F]PK14105-specific signal was of similar magnitude but peaked earlier and was retained for only 45 min. Specific signals with both ligands were also detected in regions remote from the primary lesion site, e.g., in the hippocampus and substantia nigra. Predosing animals with a large dose of PK11195 (3 mg/kg), sufficient to saturate peripheral-type benzodiazepine binding sites, abolished in vivo binding of both [3H]PK11195 and [18F]PK14105 to both primary- and remote-lesioned tissues. The specific signal with both ligands could be of sufficient magnitude and duration to make tomographic studies in humans feasible.     

Energy transfer between Chlorophyll C and Chlorophyll A

Spectral properties of solutions containing mixtures of chlorophyll a and chlorophyll c are investigated. The yield of excitation energy migration from chlorophyll c to chlorophyll a is obtained ranging from 23 to 48% dependent on the used dye concentrations. The back transfer from chlorophyll a to chlorophyll c is negligible. The shape of the polarization excitation spectrum of chlorophyll c in the Soret band region is less composed than that of chlorophyll a. Depolarization of chlorophyll a fluorescence by chlorophyll c is in agreement with the conclusion drawn from fluorescence quenching that excitation energy migrates from chlorophyll c to chlorophyll a.     

Changes of dry mass of blood lymphocytes in the course of mouse estrous cycle and their dependence on the sexual steroid hormones

The dry mass of blood lymphocytes in female mice has been found to underline the variations according to the phases of estrous cycle. Ovariectomy caused disappearance of cyclic changes and reduced the mean dry mass of lymphocytes. Whereas estradiol was effective in restoration of the lymphocyte dry mass to the values characteristic for non-operated animals, progesterone failed to produce any noticeable effect. It is concluded that estradiol is responsible for the changes of lymphocyte dry mass observed during the successive phases of estrous cycle.     

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.     

Interactions of Chlorophyll and Polypeptide Mixture with Bacterial Reaction Centres

In aqueous solutions of chlorophyll (Chl) a with synthesized polypeptides, at high ratios of Chl to polypeptides (about 75–150 µM to 500 µM) clusters of polypeptides and pigment molecules were formed. The main absorption maxima of more than one formed cluster were located at about 500 nm (Soret band) and in the region of 720–806 nm (red band). The formation of these clusters was fairly slow (some hours) at room temperature and even slower at 4 °C. The rate of cluster formation increased with the increase in Chl concentration. The addition of the even low amount of reaction centres (RCs), separated from the purple bacteria Rhodobacter sphaeroides, to the sample of Chl with polypeptides caused a very strong decrease in the efficiency of cluster formation, and a change in concentration ratios of various pigment-polypeptide aggregates. It was probably a competition between the interaction of Chl with polypeptides and with the RCs. The yield of thermal deactivation of the clusters was high, much higher than that for the RCs alone and it was different for various types of cluster. The clusters absorbing at 725–750 nm were fluorescent with maximum of emission at about 770 nm, whereas clusters absorbing at about 800 nm were nonfluorescent.     

Expression of aquaporin 1 in the pig peri-ovarian vascular complex during the estrous cycle and early pregnancy

Aquaporin 1 (AQP1) is a water channel protein expressed in endothelial and epithelial cells of many tissues, including the vasculature, where it serves to increase water permeability of the cell membrane. The aim of this study was to investigate the expression and distribution of AQP1 in porcine peri-ovarian vascular complex (PVC) during the estrous cycle and early pregnancy. Immunohistochemistry and semi-quantitative immunoblotting techniques were used. We have demonstrated the presence of AQP1 protein in the endothelial cells of the lymphatic and vascular endothelium of the PVC during the pig estrous cycle and early pregnancy. The expression of AQP1 protein in the PVC did not change significantly between Days 10-12 and 14-16, but increased on Days 2-4 and 18-20 when compared with Days 10-12 and 14-16 of the estrous cycle. In pregnant gilts, the expression of AQP1 did not differ significantly during the onset and the end of the implantation process and also when compared to the mid- and late-luteal phases of the estrous cycle. In conclusion, AQP1 is expressed in the endothelial cells of PVC and may modulate hormonal regulation of reproductive organs.