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1.
Photosystem I of the cyanobacterium Synechococcus elongatus contains two spectral pools of chlorophylls called C-708 and C-719 that absorb at longer wavelengths than the primary electron donor P700. We investigated the relative quantum yields of photochemical charge separation and fluorescence as a function of excitation wavelength and temperature in trimeric and monomeric photosystem I complexes of this cyanobacterium. The monomeric complexes are characterized by a reduced content of the C-719 spectral form. At room temperature, an analysis of the wavelength dependence of P700 oxidation indicated that all absorbed light, even of wavelengths of up to 750 nm, has the same probability of resulting in a stable P700 photooxidation. Upon cooling from 295 K to 5 K, the nonselectively excited steady-state emission increased by 11- and 16-fold in the trimeric and monomeric complexes, respectively, whereas the quantum yield of P700 oxidation decreased 2.2- and 1.7-fold. Fluorescence excitation spectra at 5 K indicate that the fluorescence quantum yield further increases upon scanning of the excitation wavelength from 690 nm to 710 nm, whereas the quantum yield of P700 oxidation decreases significantly upon excitation at wavelengths longer than 700 nm. Based on these findings, we conclude that at 5 K the excited state is not equilibrated over the antenna before charge separation occurs, and that approximately 50% of the excitations reach P700 before they become irreversibly trapped on one of the long-wavelength antenna pigments. Possible spatial organizations of the long-wavelength antenna pigments in the three-dimensional structure of photosystem I are discussed.  相似文献   

2.
The levels of alanine, aspartate and glutamine transaminase increase considerably in some diseases. We measured the activity of these enzymes and of the transaminase of 3-hydroxykynurenine, an aminoacid, which acts as a UV lens filter. Alanine and glutamine transaminases (carboxypeptidase) were not detected in normal and cataractous human lenses, and aspartate transaminase was found only in the cortex of normal lenses. 3-hydroxykynurenine transaminase was not found in lenses from persons below thirty years of age, but was found in lenses at about fifty years of age, and in cataractous lenses. Transamination of 3-hydroxykynurenine leads to the formation of xanthurenic acid and its derivatives. These substances appear to be responsible for the increase of lens fluorescence during cataract development.  相似文献   

3.
Human age-related nuclear cataract is associated with progressive and widespread oxidation of proteins, particularly in the centre of the lens. The reasons for the onset of cataract and why this disease should take place only in the lenses of older individuals remain unclear. However, a common feature of nuclear cataract is the low concentration of reduced glutathione (GSH) in the centre of the lens. GSH is the principal lenticular antioxidant of the lens and it is synthesized and regenerated in the lens cortex. In this study we investigated the diffusion of glutathione within the human lens as a function of age. Normal human lenses were incubated in artificial aqueous humor containing [35S]cysteine and the label was metabolically incorporated into GSH. After 48-h incubation, lenses were sectioned and phosphorimaging was used to determine the distribution of 35S label. In young lenses, label appeared to diffuse uniformly throughout the whole lens. By contrast, in lenses over the age of 30, very little 35S had penetrated to the centre of the lens. A distinct zonal pattern of label distribution was noted in the older lenses after 48 h incubation, which had dimensions of approximately 7.2 mm (diameter) by 2.8 mm (axial). In some older lenses this pattern was noticeable even after 96-h incubation. Thus a barrier to the diffusion of GSH was observed in older normal lenses which was not present in younger lenses. Furthermore, the internal zone thus delineated has dimensions that coincide with those of the coloured and sclerotic zone present in nuclear cataract lenses. Since nuclear cataract is a disease of the elderly, and maintenance of GSH is known to be vital for lens clarity, we propose that the development of a barrier to the movement of GSH from its site of synthesis and regeneration in the cortex, into the nucleus in older normal lenses, may over time allow oxidative modification of protein to take place in the nucleus, resulting ultimately in nuclear cataract.  相似文献   

4.
Dynamics of proteins and membranes are usually investigated by red-edge excitation spectra and fluorescence anisotropy. In a viscous or rigid medium, the fluorescence maximum position changes with the excitation wavelength upon red-edge excitation. In addition to the shift in the emission maximum on red edge excitation, fluorescence anisotropy is also known to be dependent on the excitation and emission wavelengths in viscous media. However, this dependence has always been explained by the fact that the fluorophore is rigid, i.e. it does not display any residual motions. The aim of the present work was to check the validity of this latest assumption and to explain the possible origin of the dependence of the anisotropy on both the excitation and emission wavelengths. Therefore, we compared the results obtained from the fluorescence of the Trp residues of two alpha 1-acid glycoproteins (orosomucoid). One protein was purified by chromatographic methods (orosomucoid(c)) and the other was obtained with ammonium sulfate precipitation (orosomucoid(s)). Trp residues of orosomucoidc display free motions while those of orosomucoids are rigid. The general qualitative feature of the excitation anisotropy spectra recorded on both types of preparation is identical and resembles that obtained for other proteins containing tryptophan residue in protein. The fluorescence anisotropy measured across the emission spectra decreases for both preparations, indicating that this phenomenon is characteristic for fluorophores surrounded by a rigid microenvironment or by a microenvironment that displays motions. The fluorescence anisotropy variation across the emission and the excitation spectra is more important when the fluorophore possesses constrained motions than when it displays a high degree of freedom. Our results clearly demonstrate that the tertiary structure of the protein and the structure and dynamics of the microenvironments of the Trp residues are the origin of the dependence of anisotropy on the excitation and emission wavelengths.  相似文献   

5.
We measured a.c. admittances for isolated rat lenses over a wide frequency range of 100 Hz to 500 MHz. In order to evade electrode polarization at low frequencies, we examined extracted lenses by direct semi-squeezing with Pt black-coated platinum electrodes. This gave rise to a 10(6) increment in permittivity at low frequencies, and two distinct dielectric dispersions were exhibited; dispersion 1 with a characteristic frequency (fc) of 2 kHz, and dispersion 2 with an fc of 2 MHz. In terms of loss tangent function, the two dispersions clearly dominated as two peaks. By curve fitting analysis with an allocated ellipsoidal-shells model based on ultrastructural features of the lens with a lens equivalent circuit, dispersion 1 was assigned to the equatorial cortex where regularly arranged lens fibers ran parallel to the applied electric field, and dispersion 2 to the nucleus of complicated fibers, combined with the polar cortex where lens fibers ran perpendicular to the electric field. Our results suggested that both lens dispersions were beta-dispersions attributable to bio-membranes.  相似文献   

6.
A case of hemispheric infarction involving the territory of the right middle cerebral artery and the thalamus showed conspicuous asymmetric degeneration in the substantia nigra, red nuclei, inferior olivary nuclei and dentate nuclei with concomitant changes of progressive supranuclear palsy (PSP). The right substantia nigra and red nucleus showed loss of neurons and proliferation of astrocytes. The right olivary nucleus was hypertrophic, while the neuronal loss and astrocytosis in the dentate nucleus were predominant on the contralateral side. Modified Gallyas-Braak staining revealed the extensive distribution of neurofibrillary tangles (NFTs), threads and intraglial argyrophilic structures in the globus pallidus, subthalamic nuclei, cerebral cortex and dentate nuclei, as well as in the affected brain stem nuclei, with a distinct predominance on the affected side. In this case, the one-sided predominance of the extended degeneration in these brain stem and cerebellar areas is considered, in addition to the PSP changes, to be due to secondary retrograde degeneration via the nigrostriatal and dentato-rubro-thalamic pathways following the hemispheric infarction, and to also be the result of disruption of the dentato-olivary fiber connections. In addition, because of the predominant distribution of NFTs on the more degenerated side, it is surmised that the formation of NFTs may be accelerated by secondary degeneration.  相似文献   

7.
Pathologic findings are presented in a case of neurodegenerative disease with a 5-year course of progressive presenile dementia and parkinsonism. At autopsy, atrophy of the cerebral cortex, caudate nucleus, and substantia nigra were observed. Homogeneous eosinophilic intracytoplasmic neuronal inclusions were found, predominantly in the brainstem and cerebellar nuclei. These inclusions were nonargyrophilic and did not stain with antibodies against tau and ubiquitin.  相似文献   

8.
BACKGROUND: Two to four percent of epileptics have their seizures triggered by flickering light, an effect which may be wavelength-dependent. We evaluated a patient with a long-standing history of light-induced petit mal seizures to determine if the seizures were triggered more effectively by a particular range of wavelengths and to determine whether this information could be used in the optometric management of such patients. METHODS: Flickering lights of different wavelengths but equal luminance were presented while the patient's electroencephalogram (EEG) and subjective reports were monitored. RESULTS: The EEG results were not significantly different for different wavelengths, but the patient reported that longer wavelength light induced stronger seizures more consistently. Based on these results we concluded that the patient might benefit from spectacles tinted to exclude transmission of red light. The patient was given 4 pairs of 85% transmission spectacles, which differed only in lens tint (red, yellow, green, and blue), to try over a period of time. The patient felt that the tinted lenses were not dark enough to decrease his seizures effectively and he has opted to wear standard sunglasses. CONCLUSIONS: We feel this case shows that patients with light-induced seizures can benefit from optometric consultation. Because the patient's subjective report identified the clearest wavelength effect, we feel that it is reasonable for the general practitioner to suggest deeply tinted lenses to reduce the frequency of seizures in these patients.  相似文献   

9.
During the past decade, ambient ultraviolet radiation has been implicated in the age-related increase in fluorescence and pigmentation of the human lens nucleus. 8-Methoxypsoralen (8-MOP) (currently in vogue for the treatment of psoriasis) is a well-known photosensitizing agent. This drug lar fluorescence (360/440 nm.) and a change in receiving a single intraperitoneal dose (4 to 8 mg./kg.) of 8-MOP. When such rats are subjected to ambient light or ultraviolet (UV) radiation in vivo, there is an enhancement of lenticular fluorescence (360/440 nm.) and a change in their phosphorescence spectra. In vitro studies on lenses derived from rats given 8-MOP and exposed to monochromatic UV radiation show effects similar to those of the in vivo experiments. The foregoing studies demonstrate that 8-methoxypsoralen enters the lens and can be affected by ambient light as well as UV radiation, resulting in a photosensitized enhancement of lenticular fluorescence and a binding of this photosensitizing agent to macromolecules within the lens.  相似文献   

10.
It has been established that hippocampus, enthorhinal cortex, amygdala and substantia nigra (pars reticulata) lesions before head injury lead to a decrease of kainic acid-induced behavioral and electrographic seizure expressions. It can be concluded that after head injury the activation of limbic structures excitability due to excitation of "inputs" to these formations takes place. The obtained data indicate the significant role of nucleus caudatus in activation of posttraumatic brain excitatory mechanisms.  相似文献   

11.
PURPOSE: To visualize the structure and determine the continuity of lipid membranes in lens fiber cells (LFCs) from human aged normal and cataractous lenses. METHODS: Thick sections from human nuclear cataracts and aged normal lenses were stained with the lipophilic probe DiI, and then analyzed by confocal microscopy. Staining patterns of membranes were observed in individual optical sections or three-dimensional projections of z-series taken in longitudinal section and cross-section of LFCs from different regions within the lens nucleus. RESULTS: DiI bound to and delineated the plasma membrane of LFCs from all regions of the lens nucleus. Three-dimensional projections of z-series from aged normal and cataractous lenses suggested that some of the stained lipid membranes were not continuous with LFC plasma membrane of cataractous lenses. CONCLUSIONS: The results obtained using these methods demonstrated that lipid membranes, discontinuous with the plasma membrane of LFCs, were indicative of a novel process occurring predominately in cataractous human lenses.  相似文献   

12.
We have quantified the influence of 'Y' sutures on lens optical quality (spherical aberration, i.e. focal length variability) as a function of age. Young (n = 6) and old (group 1, n = 5; and group 2, n = 4) bovine lenses were initially scanned by a low-power (2 mW) helium-neon laser beam passed either through or at a series of acute angles to suture branches. In all lenses, focal length variability was least when the beam was passed through areas of the lens devoid of sutures and greatest when passed through sutures. In older lenses, variability was also significantly increased in all locations though to a greater degree at sutures. Correlative morphological analysis by scanning electron microscopic (SEM) and three dimensional (3-D) computer-assisted drawings (CADs) revealed the following: (1) young lenses had uniformly hexagonal fibers arranged in parallel radial cell columns (RCCs), while old lenses had nonuniformly hexagonal fibers arranged in variably parallel RCCs; (2) the irregularly-sized and -shaped ends of young fibers overlapped within growth shells to form complementary anterior and posterior symmetrical 'Y' suture patterns, while larger and more irregularly-shaped ends of older lenses overlapped to form asymmetrical 'Y' patterns; and (3) the identical suture patterns in successive shells of young lenses resulted in inverted triangular suture planes extending from the embryonic nucleus to the lens periphery, while the progressively wider and more serpentine suture branches of old lenses resulted in inverted pyramidal suture planes with narrow apices oriented towards the embryonic nucleus and broad irregular bases oriented toward the lens periphery. Thus, there is a significant interrelationship between lens optical quality and structure that varies as a function of age. These results extend and confirm the results of our earlier studies on lenses with simpler 'line' sutures and preface our studies of primate lenses with more complex 'star' sutures. All of these studies show that lens sutural anatomy should be considered when evaluating the optical quality of normal and pathological lenses.  相似文献   

13.
PURPOSE: To establish whether advanced glycation is the major mechanism for yellowing of lens proteins. METHODS: Synchronous fluorescence (SF) and immunochemical assays were used to study glycation in vitro and in vivo. In the in vitro study, advanced glycation end products (AGEs) were prepared and used as antigens to induce antibodies to AGEs. The in vitro AGEs and classified nuclear cataracts were analyzed by SF and immunochemical assays. RESULTS: In vitro AGEs generated from various glycating agents and carrier proteins displayed strong SF above 350 nm; the spectra were well resolved with major bands at 380 nm and 420 nm. Samples from human lenses manifested a band at 395 nm in addition to the two bands shown by in vitro AGEs. SF intensity is greater for the water-insoluble (WI) than water-soluble (WS) fraction, but both increased with increasing nuclear color. The immunoreactivity data also showed that the WI fraction contained more AGEs than the WS fraction and that the amount of AGEs increased with increasing nuclear color. CONCLUSIONS: Fluorescence and immunoassays indicated that pigmented AGEs contributed to yellowing of the crystalline lens nucleus.  相似文献   

14.
In view of renewed interest in the lens epithelium as the initiation site for cataract development, it seemed timely to review recent studies which appear to establish UV damage in the lens epithelium as the cause of UV cataract. While UV photons can and do interact with lens proteins in the cortex and nucleus, experimental results from cultured lenses and tissue cultured epithelial cells also demonstrate both mutagenic and cytotoxic effects in the epithelium. This minireview examines UV-induced changes in lens physiology that appear to follow epithelial cell damage, including inactivation of critical enzymes of transport and metabolic processes. Changes in membrane function include altered cation transport, increased permeability, and altered biosynthesis. One potential scenario for the propagation of damage from the epithelium to the underlying fiber cells includes calcium elevation, an early event in cataract development and critical to many physiological processes.  相似文献   

15.
Angiotensin IV (Val Tyr Ile His Pro Phe), administered centrally, increases memory retrieval and induces c-fos expression in the hippocampus and piriform cortex. Angiotensin IV binds to a high affinity site that is quite distinct in pharmacology and distribution from the angiotensin II AT1 and AT2 receptors and is known as the AT4 receptor. These observations suggest that the AT4 receptor may have multiple central effects. The present study uses in vitro receptor autoradiography, and employs [125I]angiotensin IV to map AT4 receptors in the macaca fascicularis brain. The distribution of the AT4 receptor is remarkable in that its distribution extends throughout several neural systems. Most striking is its localization in motor nuclei and motor associated regions. These include the ventral horn spinal motor neurons, all cranial motor nuclei including the oculomotor, abducens, facial and hypoglossal nuclei, and the dorsal motor nucleus of the vagus. Receptors are also present in the vestibular, reticular and inferior olivary nuclei, the granular layer of the cerebellum, and the Betz cells of the motor cortex. Moderate AT4 receptor density is seen in all cerebellar nuclei, ventral thalamic nuclei and the substantia nigra pars compacta, with lower receptor density observed in the caudate nucleus and putamen. Abundant AT4 receptors are also found in areas associated with cholinergic nuclei and their projections, including the nucleus basalis of Meynert, ventral limb of the diagonal band and the hippocampus, somatic motor nuclei and autonomic preganglionic motor nuclei. AT4 receptors are also observed in sensory regions, with moderate levels in spinal trigeminal, gracile, cuneate and thalamic ventral posterior nuclei, and the somatosensory cortex. The abundance of the AT4 receptor in motor and cholinergic neurons, and to a lesser extent, in sensory neurons, suggests multiple roles for the AT4 receptor in the primate brain.  相似文献   

16.
An apyrase (NTP/NDPase) implicated in the response of Mimosa pudica to stimuli, such as touch, has been cloned, sequenced and expressed in Escherichia coli. While purifying and characterizing this enzyme, it was observed that a chromophore is associated with it, having absorption in the ultraviolet-A/blue region of the spectrum. The absorbance maximum of the chromophore, purified from the enzyme complex by gel filtration and HPLC, is around 350 nm. The chromophore has been identified as N5,N10-methenyl tetrahydrofolate (MTHF) by comparing the excitation and emission spectra of synthetic MTHF and the isolated cofactor, and by reconstitution of the enzyme complex with synthetic MTHF. Upon excitation with light (350 nm), an increase of apyrase activity was observed in the purified or reconstituted holoenzyme but not in the apoenzyme. The wavelength dependence of the light stimulation matched well with the fluorescence excitation spectra of the cofactor, MTHF. Possible implications of the results for signal transduction in M. pudica have been discussed.  相似文献   

17.
To understand better how the brainstem may influence thalamocortical activity, we have examined the projection patterns of different brainstem nuclei to the thalamic reticular nucleus. Iontophoretic injections of biotinylated dextran were made into various nuclei of the brainstem (superior colliculus, periaqueductal grey matter, parabrachial nucleus, pedunculopontine tegmental nucleus, laterodorsal tegmental nucleus, substantia nigra, ventral tegmental area, and locus coeruleus) of Sprague-Dawley rats by using stereotaxic coordinates. Our results show that afferents from each brainstem nucleus make distinct zones within the reticular nucleus. For example, the superior colliculus projects largely to the dorsal parts of the reticular nucleus, whereas the pedunculopontine nucleus projects to the ventral parts of the reticular nucleus. The substantia nigra, on the other hand, projects to the ventrolateral edge of the reticular nucleus. We also examined the distribution of these brainstem afferents within the dorsal thalamus and compared these distributions with those seen in the reticular nucleus. We found three different patterns. First, a given brainstem nucleus projects to a particular dorsal thalamic nucleus as well as to the corresponding, functionally associated, reticular sector (e.g., from the substantia nigra). Second, a given brainstem nucleus projects to a particular dorsal thalamic nucleus but not to the corresponding reticular sector (e.g., from the superior colliculus). Finally, a given brainstem nucleus projects to a given reticular sector but not to the corresponding dorsal thalamic nucleus (e.g., from the midbrain reticular nucleus). In general, our results indicate that various brainstem nuclei project to particular territories of the thalamic reticular nucleus. Through these reticular projections, brainstem nuclei may influence distinct thalamocortical pathways in addition to those that are influenced by their direct projection to the dorsal thalamus.  相似文献   

18.
The expression of cadherin-8 was mapped by in situ hybridization in the embryonic and postnatal mouse central nervous system (CNS). From embryonic day 18 (E18) to postnatal day 6 (P6), cadherin-8 expression is restricted to a subset of developing brain nuclei and cortical areas in all major subdivisions of the CNS. The anlagen of some of the cadherin-8-positive structures also express this molecule at earlier developmental stages (E12.5-E16). The cadherin-8-positive neuroanatomical structures are parts of several functional systems in the brain. In the limbic system, cadherin-8-positive regions are found in the septal region, habenular nuclei, amygdala, interpeduncular nucleus, raphe nuclei, and hippocampus. Cerebral cortex shows expression in several limbic areas at P6. In the basal ganglia and related nuclei, cadherin-8 is expressed by parts of the striatum, globus pallidus, substantia nigra, entopeduncular nucleus, subthalamic nucleus, zona incerta, and pedunculopontine nuclei. A third group of cadherin-8-positive gray matter structures has functional connections with the cerebellum (superior colliculus, anterior pretectal nucleus, red nucleus, nucleus of posterior commissure, inferior olive, pontine, pontine reticular, and vestibular nuclei). The cerebellum itself shows parasagittal stripes of cadherin-8 expression in the Purkinje cell layer. In the hindbrain, cadherin-8 is expressed by several cranial nerve nuclei. Results from this study show that cadherin-8 expression in the embryonic and postnatal mouse brain is restricted to specific developing gray matter structures. These data support the idea that cadherins are a family of molecules whose expression provides a molecular code for the regionalization of the developing vertebrate brain.  相似文献   

19.
The present study determined the effect of bilateral lesions of specific cortical or thalamic nuclei that provide excitatory amino acid afferents to the nucleus accumbens (i.e. the dorsal prefrontal cortex, ventral prefrontal cortex, amygdala, hippocampus and periventricular thalamus) on the expression of cocaine-induced behavioral sensitization. Lesions of these nuclei were made during a three-week withdrawal period following repeated daily injections of cocaine or saline. The results indicate that dorsal prefrontal cortex lesions block the expression of behavioral sensitization to cocaine, while ventral prefrontal cortex, fimbria fornix, amygdala and thalamic lesions have no effect. A subsequent microdialysis experiment was performed in order to evaluate the effect of dorsal prefrontal cortex lesions on glutamate transmission in the nucleus accumbens core of cocaine- and saline-pretreated rats. The systemic injection of cocaine produced a significant increase in extracellular glutamate in the nucleus accumbens core among animals with a sham surgery; this effect was blocked by a bilateral lesion of the dorsal prefrontal cortex. Taken together, these results indicate that the dorsal prefrontal cortex, which provides excitatory amino acid input selectively to the core region of the nucleus accumbens, enhances the expression of behavioral sensitization to cocaine by increasing glutamate transmission in this subnucleus.  相似文献   

20.
To understand better the organisation of zona incerta of the thalamus, this study has examined the patterns of connections that this nucleus has with various nuclei of the brainstem. Injections of biotinylated dextran or cholera toxin subunit B were made into the dorsal raphe, midbrain reticular nucleus, pedunculopontine tegmental nucleus, periaqueductal grey matter, pontine reticular nucleus, substantia nigra, superior colliculus, and ventral tegmental area of Sprague-Dawley rats, and their brains were processed by using standard tracer-detection methods. In general, our results show that zona incerta forms the major zone in the thalamus where these ascending brainstem axons terminate and from which descending axons that travel back to these same brainstem centres originate. These incertal inputs and outputs are limited largely to a distinct sector of zona incerta, the dorsal sector. An exception to this pattern is evident in the incertal projection to the deep layers of the superior colliculus; this projection, unlike all of the others, arises from cells in the ventral sector of zona incerta. Our results also show little evidence for a well-defined topography of projection between the brainstem and the zona incerta. For instance, small injections into each brainstem nucleus result in labelled terminals and in cells spread throughout much of the dorsal sector of zona incerta, with no local zone of concentration within the sector. Again, an exception to this pattern is seen in the incertal projection to the superior colliculus. This projection, unlike the others, shows a clear topographical organisation: A medial-lateral shift in the injection site in the colliculus results in a lateral-medial shift in the position of labelled cells in zona incerta. Curiously, even though the incertal projection to the colliculus appears to be mapped, the collicular projection back to zona incerta is not mapped. In conclusion, then, a number of brainstem nuclei (except for the deep collicular layers) have strong and overlapping connections within the same sector of zona incerta. This convergence of many functionally diverse brainstem afferents within zona incerta places this nucleus in a strategic position to sample the general activity of the brainstem and, perhaps, acts as a relay of this information to higher centres, such as the dorsal thalamic relay nuclei and the cerebral hemispheres.  相似文献   

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