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Studies on the Mechanism of Photoinhibition in Higher Plants: I. EFFECTS OF HIGH LIGHT INTENSITY ON CHLOROPLAST ACTIVITIES IN CUCUMBER ADAPTED TO LOW LIGHT.

Cucumber plants (Cucumis sativus L.), grown at low quantum flux density (120-150 microeinsteins per square meter per second) were photoinhibited by a three-hour exposure in air to ten times the light intensity experienced during growth. Chloroplasts were isolated from photoinhibited and control leaves and the following activities determined: O(2) evolution in the presence of ferricyanide, photosystem I activity, noncyclic and cyclic photophosphorylation, and light-induced proton uptake. Chlorophyll and chloroplast absorbance spectra, and chloroplast fluorescence were also measured. It was found that photosystem II electron transport and non-cyclic photophosphorylation were inhibited by about 50%, while cyclic photophosphorylation was less inhibited and photosystem I electron transport and light-induced proton uptake were unaffected. Electron transport to methylviologen could not be fully restored by electron donation to photosystem II. Chloroplast fluorescence induction at room temperature was strongly reduced following photoinhibition. There was no difference in the absorption spectra of the extracted chlorophylls from control and photoinhibited chloroplasts, but an increase of the absorption in the blue wavelength region was observed in the photoinhibited chloroplasts. It is suggested that high light stress does not result in alteration of the membrane properties, as is the case in low-temperature stress for example, but affects directly the photosynthetic reaction centers, primarily of photosystem II.

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Effectiveness of intermittent light treatments on anthocyanin synthesis in dark-grown and light-pretreated seedlings.

Differences in the extent of anthocyanin production between intermittent light treatments with short and long dark intervals between successive irradiations are more pronounced in dark-grown than in light-pretreated cabbage seedlings. This observation is consistent with the hypothesis, based on destruction kinetics data, that there might be two pools of phytochrome, a labile one and a stable one, present in different proportions in dark-grown and light-pretreated seedlings, and suggests that light-dependent changes of the stable to labile phytochrome ratio might be physiologically significant in the photoregulation of photomorphogenic responses.

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Studies on the Photoactivation of the Water-Oxidizing Enzyme: II. Characterization of Weak Light Photoinhibition of PSII and Its Light-Induced Recovery.

Inactivation of the water splitting enzyme complex in leaves or isolated chloroplasts results in increased susceptibility of photosystem II (PSII) to damage by light. Photoinhibition under this condition occurs in very weak light. The site of damage is exclusive of the water splitting complex yet still on the oxidizing side of PSII, as the Q(B) locus is unaffected while photoreduction of silicomolybdate is inhibited. The kinetics of loss in PSII activity are more complex than apparent first-order, and the quantum efficiency is low. We observe no evidence of deletion from thylakoid membranes of any PSII polypeptide as a consequence of photoinhibition, although recovery from the photoinhibition is dependent upon both light and 70S protein synthesis. Enhanced synthesis of two proteins occurs during recovery, only one of which (D2) appears to be causally related to the recovery. We present a model which describes the relationship of weak light photoinhibition and its recovery to photoactivation of the S-state water oxidizing complex.

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Zeaxanthin and the Heat Dissipation of Excess Light Energy in Nerium oleander Exposed to a Combination of High Light and Water Stress.

Upon termination of watering of plants of Nerium oleander exposed to high light, photochemical efficiency became reduced as leaf water content decreased. Evidence is presented that this type of photoinhibition reflects to a substantial degree radiationless dissipation of excitation energy, probably mediated by the carotenoid zeaxanthin. During the imposition of water stress, the zeaxanthin content of leaves increased at the expense of violaxanthin and beta-carotene as a water deficit developed over a period of several days. The increase in zeaxanthin content was linearly related to an increase in the rate of radiationless energy dissipation in the antenna chlorophyll as calculated from the characteristics of chlorophyll a fluorescence measured with a pulse amplitude modulated fluorometer at room temperature. The increase in the rate of radiationless dissipation was also linearly related to a decrease in PSII photochemical efficiency as indicated by the ratio of variable to maximum fluorescence. Leaves of well-watered shade plants of N. oleander exposed to strong light showed a similar increase in zeaxanthin content as sun leaves of the same species subjected to drought in strong light. Shade leaves possessed the same capacity as sun leaves to form zeaxanthin at the expense of both violaxanthin and beta-carotene. The resistance of this species to the destructive effects of excess light appears to be related to interconversions between beta-carotene and the three carotenoids of the xanthophyll cycle.

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Hierarchical Response of Light Harvesting Chlorophyll-Proteins in a Light-Sensitive Chlorophyll b-Deficient Mutant of Maize.

The light-sensitive chlorophyll b (Chl b)-deficient oil yellow-yellow green (OY-YG) mutant of maize (Zea mays) grown under conditions of high light exhibits differential reductions in the accumulation of the three major Chl b-containing antenna complexes and characteristic changes in thylakoid architecture. When observed by freeze-fracture electron microscopy, the most notable changes in the OY-YG thylakoid structure are: (a) a major reduction in the number of 8 nanometer particles of the protoplasmic fracture face of stacked membrane regions (PFs) paralleled by a 60% reduction in the chlorophyll-proteins (CP) associated with the peripheral light harvesting complex (LHCII) for photosystem II (PSII) and which give rise to the LHCII oligomer/monomer (CPII(*)/CPII) bands on mildly dissociated green gels; (b) a sizable decrease in the proportion of 11 to 13 nanometer particles of the protoplasmic fracture face of unstacked membrane regions (PFu) that parallels the loss of light harvesting complex I (LHCI) antennae from photosystem I (PSI) centers and a 40% reduction of the band containing CP1 and LHCI (CPI(*)) on mildly dissociating green gels; (c) an unchanged or slightly increased average size of particles of the exoplasmic fracture face of stacked (or appressed) membrane regions (EFs) along with a relative increase in CP29, the postulated bound LHC of PSII, and of CP47 and CP43, PSII core antenna complexes. This latter result sets the OY-YG mutant apart from all other Chl b-deficient mutants studied to date, all of which possess EFs particles that are substantially reduced in size. Based on these findings, we postulate that the bound LHCII associated with EFs particles consists mostly of CP29 chlorophyll proteins and very little, if any, CPII(*)/CPII chlorophyll proteins. Indeed, the CPII(*)/CPII chlorophyll proteins may be exclusively associated with the ;peripheral' LHCII units that give rise to 8 nanometer PF particles. The differential effect of the Chl b deficiency on the accumulation of the three main antenna complexes (CPII(*)/CPII>CPI(*)>CP29) suggests, furthermore, that there is a hierarchy among Chl b-binding proteins, and that this hierarchy might be an integral part of long-term photoregulation mediating Chl b partitioning in the chloroplast.

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Light intensity regulates the accumulation of the major light-harvesting chlorophyll-protein in greening seedlings.

Etiolated pea (Pisum sativum [L.] cv Progress 9) and barley (Hordeum vulgare [L.] cv Boone) seedlings greened under either low (40 microeinsteins per square meter per second) or high (550 microeinsteins per square meter per second) intensity light were analyzed for chlorophyll (Chl) content and the levels of mRNA and protein for the major light-harvesting chlorophyll (Chl)-protein of photosystem II (LHC-II). Low intensity plants accumulated Chl more rapidly than high intensity plants. Both single radial immunodiffusion analysis and mild sodium dodecyl sulfate-polyacrylamide gel electrophoresis green gels showed that low intensity plants also accumulated LHC-II protein more rapidly than high intensity plants, following a kinetic pattern similar to the total Chl data. In contrast, LHC-II mRNA levels appeared to be independent of LHC-II protein levels although pea and barley LHC-II mRNA exhibited different light intensity responses. The absence of coordination between LHC-II mRNA and protein levels suggested that the biosynthesis of LHC-II in greening seedlings is not limited by mRNA. A correlation (better than the 0.01 significance level) between LHC-II protein accumulation and Chl accumulation was found for both pea and barley. The accumulation of LHC-II protein was not linked to the development of photosynthetic electron transport. These results and the similar effect of light intensity on Chl content and LHC-II protein levels suggested that the availability of Chl may limit LHC-II protein accumulation in greening seedlings.

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Reflected Far-Red Light Effects on Chlorophyll and Light-Harvesting Chlorophyll Protein (LHC-II) Contents under Field Conditions.

The influence of various colors of soil cover (mulch) on the farred/red (FR/R) ratio in upwardly reflected light and on concentrations of chlorophyll (Chl) and light-harvesting Chl protein (LHC-II) were measured under field conditions. The FR/R ratios above green surfaces were higher than over white surfaces. Even though plants (Gossyplum hirsutum L. cv PD-1) were grown in full sunlight, those that received higher FR/R ratios in upwardly reflected light were taller and had thinner leaves with higher concentrations of Chl and LHC-II. A controlled environment experiment showed FR/R control of Chl and LHC-II concentrations. The results illustrate the importance of spectral distribution of reflected light on plant growth and a potential means of altering the chemistry of leaf crops under field conditions.

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Interactions Between Light and Gas Vacuoles in Halobacterium salinarium Strain 5: Effect of Ultraviolet Light.

The potential light shielding by intracellular gas vacuoles in Halobacterium salinarium strain 5 was examined by looking at the ultraviolet light inactivation curves of both wild-type cells and mutants which are defective in the production of gas vacuoles. Whereas strains defective in gas vacuole production were slightly more sensitive to ultraviolet inactivation, no significant differences in ultraviolet sensitivity were seen, indicating that these subcellular inclusion bodies are not effective as light-shielding organelles. In addition, it was shown that ultraviolet light acts as a plasmid-curing agent in Halobacterium.

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Phase-shifting digital holography with a low-coherence light source for reconstruction of a digital relief object hidden behind a light-scattering medium.

Hiding image data with a light-scattering medium is effective as a basic data protection technique. The hidden image data can be observed only by using a low-coherence interference technique and is thus protected from unauthorized access. Unlike an intensity-distributed object, a digital relief object has no intensity distribution, making it possible to hide its existence by using a dilute light-scattering medium. To reconstruct the digital relief object through the light-scattering medium, we developed phase-shifting digital holography with a low-coherence light source. The experimental performance, including the spatial resolution and phase error of the reconstructed image, is estimated.

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Phase unwrapping method for three-dimensional stress analysis by scattered-light photoelasticity with unpolarized light.

In scattered-light photoelasticity with unpolarized light, the secondary principal stress direction psi and the relative phase retardation rho in a three-dimensional stressed model with rotation of the principal stress axes can be obtained by use of Stokes parameters. For completely automated stress analysis, measurements of the total relative phase retardation and the secondary principal stress direction over the entire field are required, and it is necessary to unwrap psi and rho. A phase unwrapping method is thus proposed for the determination of these values based on scattered-light photoelasticity. The values are easily obtained via an arctangent function, overcoming the error associated with the quarter-wave plate by employing an incident light of different wavelengths. The proposed technique provides automated and nondestructive determination of the total relative phase retardation and the secondary principal stress direction in a model exhibiting rotation of the principal stress axes.

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Evaluation of the photosensitizer Tookad for photodynamic therapy on the Syrian golden hamster cheek pouch model: light dose, drug dose and drug-light interval effects.

We have evaluated the efficacy of the new photosensitizer (PS) Tookad in photodynamic therapy (PDT) in vivo. This PS is a palladium-bacteriopheophorbide presenting absorption peaks at 762 and 538 nm. The light dose, drug dose and drug injection-light irradiation interval (DLI), ranging between 100 and 300 J/cm2, 1 and 5 mg/kg and from 10 to 240 min, respectively, were varied, and the response to PDT was analyzed by staging the macroscopic response and by the histological examination of the sections of the irradiated cheek pouch. The level of PDT response, macroscopically and histologically, shows a strong dependence on the DLI, light dose and drug dose at the applied conditions in the normal hamster cheek pouch. A decay of the tissular response with increasing DLI is observed corresponding to a time of half-maximum response ranging from 10 to 120 min, depending on drug dose and light dose. The tissues affected at the lowest doses are predominantly the vascularized diffuse connective tissue situated between the inner and outer striated muscle (SM) layers as well as these muscle layers themselves. The highest response at the shortest DLI and the absence of a measurable response at DLI longer than 240 min at 300 J/cm2 and drug dose of 5 mg/kg are characteristics of a predominantly vascular effect of this PS. This observation suggests that Tookad could be effective in PDT of vascularized lesions or pathologies associated with the proliferation of neovessels.

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Effect of carbon dioxide on the collection of adult Culicoides spp. (Diptera: Ceratopogonidae) by a new modification of black light New Jersey light traps.

Comparisons were made between black light-modified New Jersey traps with and without dry ice as a carbon dioxide bait. The black light trap with CO2 collected significantly more nulliparous and parous empty Culicoides variipennis than the black light trap without CO2. No significant difference was detected in the number of gravid, blood engorged or male C. variipennis collected by the 2 trap types. The baited trap collected a larger proportion of nulliparous females than the black light trap. No difference in the proportion of males or other female parity groups was detected between the 2 trap types. The total number or proportion of each parity group of C. stellifer was not significantly different between the 2 trap types.

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[Therapy of acne with visible light. Decreased irradiation time by using a blue-light high-energy lamp].

Lipophilic propionibacterium acnes stores large amounts of porphyrins. This peculiarity can be used for photodynamic destruction of propionibacteria. After repeated irradiations with a blue light-type high pressure lamp (Narva NC 1000-64) both acne and seborrhoea was improved markedly (10 irradiations, each for 10 minutes, cumulative light dose 325 J/cm2). Also porphyrin content inside acne follicles dropped. Serial irradiations with short range visible light (400 to 420 nm) are recommended for acne treatment during the light-poor season of the year.

Acne Vulgaris↗

The pupillary light reflex in diabetes mellitus: evaluation of a newly developed infrared light reflection method.

Pupillary light reflexes were measured in 18 diabetic patients without clinical signs of neuropathy and in 18 control subjects, using a newly developed infrared light reflection technique called IRIS. This method has some important advantages. Apart from being non traumatic, it enables the recording of dynamic pupillary light reflexes simultaneously in both eyes. Furthermore, it has a large amplitude and time resolution (1 ms), permitting accurate determination of latency values. It was found that 78% of the diabetic patients show a significantly prolonged latency of the constriction reaction, while 39% of the diabetic patients show an abnormally prolonged dilatation latency. All patients with an abnormally prolonged dilatation latency also show a prolonged constriction latency. This finding suggests that in diabetes mellitus the parasympathetic system is affected before abnormalities of the sympathetic nervous system can be demonstrated. Assessment of the pupillary light reflex, using the sensitive IRIS method, is an easily applicable and reliable method for detection of subclinical autonomic nervous system dysfunction associated with diabetes mellitus. The method may also provide an objective tool for the assessment of therapy results in diabetic neuropathy.

Adult↗

[Serotonin binding activity of light and heavy synaptosomes from different animal brain structures following light deprivation].

The extent of interaction between serotonin (ST) with light and heavy synaptosomes from different brain regions of normal and dark-reared rabbits was studied by the author's technique, using gel filtration. The activity of this interaction in the heavy synaptosomes of the visual cortex and the colliculi superiores of the control rabbits was the same and significantly higher than that in the motor cortex. The light synaptosomes of all the investigated regions were similar by the extent of this process activity. In dark-reared rabbits the activity of ST binding by heavy synaptosomes in the visual cortex and the colliculi superiores decreased by 74--81 per cent only. A fall in the ST binding by light synaptosomes in all the regions was the same (approximately 73 per cent as compared to the control). The results obtained are discussed in the light of a possible mediator or modulator role of ST in the visual system structures.

Animals↗

Light chain loss and reexpression leads to idiotype switch. Surrogate light chains are probably responsible for this process.

The murine B-lymphocyte cell line 38C-13 is characterized by several cell surface markers typical for an early stage of B-cell differentiation. Cells of this cell line possess cell surface membrane IgM molecules composed of mu and kappa polypeptide chains. They also produce "surrogate" or "pseudo" light chains (psi L) coded by the lambda 5 and VpreB genes. Variants of the 38C-13 cell line which do not synthesize kappa chains can be isolated from the 38C-13 population by the use of anti-idiotype antibodies in vivo and in vitro. In some kappa chain-deficient variant cell lines, cells which have regained surface IgM expression but have lost the original idiotype specificity, can be isolated. This idiotype switch is probably due to a secondary rearrangement of the kappa gene. In the kappa chain-deficient variant cells, the mu chains assemble with the surrogate light chains but the assembled IgM-like molecules are not expressed on the cell surface. It is suggested that surrogate light chains play an important role in the induction of kappa gene rearrangement but that surface expression of mu-psi L complexes is not required for this process.

Animals↗

Let there be "more" light: enhancement of light actions on the circadian system through non-photic pathways.

Circadian rhythms are internally generated circa 24 h rhythms. The phase of the circadian pacemaker in mammals can be adjusted by external stimuli such as the daily cycle of light, as well as by internal stimuli such as information related to the physiological and behavioral status of the organism, collectively called "non-photic stimuli". We review a large number of studies regarding photic-non-photic interactions on the circadian system, with special focus on two widely described neurotransmitters associated with non-photic input pathways: neuropeptide Y (NPY) and serotonin 5-HT. Both neurotransmitters are capable of phase advancing the master pacemaker oscillation when applied during the subjective day, as do several behavioral manipulations. Also, both are capable of inhibiting light-induced phase shifts during the subjective night, suggesting a dynamic interaction between photic and non-photic stimuli in the fine-tuning of the pacemaker function. Suppression of the NPYergic and/or serotonergic non-photic input pathways can in turn potentiate the phase-shifting effects of light. These findings pose new questions about the possibility of a physiological role for the dynamic interaction between photic and non-photic inputs. This might be particularly important in the case of circadian system adjustments under certain conditions, such as depression, shift work or jet lag.

Animals↗

Regulatory proteolysis of the major light-harvesting chlorophyll a/b protein of photosystem II by a light-induced membrane-associated enzymic system.

An endogenous proteolytic activity associated with spinach chloroplast thylakoid membranes has been identified. This enzymic activity is involved in the degradation of the major light-harvesting chlorophyll a/b protein of photosystem II (LHCII) in response to exposure of leaves to increased irradiance. This proteolysis of LHCII requires an induction period and can only be detected 48-72 hours after transfer of the plants from low-intensity to high-intensity light. Once initiated by high-intensity light, the degradation of LHCII can readily occur in complete darkness. The proteolysis can, after induction in vivo, be experimentally followed in vitro, both in isolated intact chloroplasts and thylakoid membranes. The proteolytic process is strictly dependent on ATP and the protease involved is of the serine or cysteine type. The activity can be released from isolated thylakoid membranes by washing with high concentrations of NaCl and reconstituted by readdition of the desalted wash supernatant. It is concluded that the protease is extrinsically bound to the outer surface of the stroma-exposed regions of the stacked thylakoid membrane. The mechanism for the induction of the proteolytic process as well as its relation to previously described thylakoid proteases will be discussed.

Adenosine Triphosphate↗