[Peribulbar anesthesia: efficacy of a single injection with a limited local anesthetic volume]. / Anesthésie péribulbaire: efficacité d'une seule injection et d'un volume d'anesthésiques locaux limité.

INTRODUCTION: Cataract surgery can be performed with peribulbar anesthesia. The classical technique consists of two injections of local anesthetics. The purpose of our study was to assess peribulbar anesthesia with a single injection and a limited volume of local anesthetics. MATERIAL AND METHOD: After local ethics committee agreement and oral consent, patients scheduled for cataract surgery using peribulbar anesthesia were prospectively included. The lower temporal puncture was performed with a peribulbar needle with propofol sedation. The mixture of local anesthetics was administered with tactile control of orbital pressure. The puncture was followed by a 10-min compression of the ocular globe. Akinesia, analgesia, complications, and surgical conditions were noted. RESULTS: A total of 101 successive patients were included. We administered 1.2 mg/kg of propofol. The volume of local anesthetics administered was 5.0 +/- 0.9 ml. Ninety patients had akinesia at 10 min and 6.7% moderate chemosis. No puncture complication occurred. At the end of surgery, the pain noted by the patients was 0.4 +/- 2.1 out of 100 (range, 0-10). Surgical conditions were good for all patients. CONCLUSION: Peribulbar anesthesia performed with a single injection and a limited volume of local anesthetics allows cataract surgery in good conditions for the surgeon with very good analgesia for the patient.

Unexpected changes in photosystem I function in a cytochrome c6-deficient mutant of the cyanobacterium Synechocystis PCC 6803.

Cytochrome c6, the product of the petJ gene, is a photosynthetic electron carrier in cyanobacteria, which transfers electrons to photosystem I and which is synthesised under conditions of copper deficiency to functionally replace plastocyanin. The photosystem I photochemical activity (energy storage, photoinduced P700 redox changes) was examined in a petJ-null mutant of Synechocystis PCC 6803. Surprisingly, photosystem I activity in the petJ-null mutant grown in the absence of copper was not much affected. However, in a medium with a low inorganic carbon concentration and with NH4+ ion as nitrogen source, the mutant displayed growth inhibition. Analysis showed that, especially in the latter, the isiAB operon, encoding flavodoxin and CP43', an additional chlorophyll a antenna, was strongly expressed in the mutant. These proteins are involved in photosystem I function and organisation and are proposed to assist in prevention of overoxidation of photosystem I at its lumenal side and overreduction at its stromal side.

Light-induced carbon dioxide and oxygen uptake in plant leaves measured by the photoacoustic method.

Leaf discs, enclosed in a photoacoustic (PA) chamber, generate two types of PA gas-uptake signals under certain conditions. Type I is manifested by a severe signal decrease that develops slowly under very low-light intensity and often reaches negative values. It is partially reversed by low-intensity far-red light. Type II occurs transiently in modulated far-red light. It is manifested by a rapid and dramatic decrease of the PA signal, upon the addition of short-wave background light, which is subsequently reversed. It differs from type-I uptake in that it occurs at much higher total light intensities. A thorough study, including modulation frequency and atmospheric composition dependencies, indicates different mechanisms for the two types of uptakes. Type-I uptake results from CO2 accumulation in the PA cell by leaf respiration and reflects modulations in CO2 solubilization. Type-II uptake likely reflects oxygen photoreduction in photosystem I, occurring prior to the activation of photosynthesis (i.e. during photosynthesis induction). This is supported by the complete suppression of type-II uptake when O2 was removed. Also, type-II uptake was only mildly sensitive to CO2 elimination, whereas type-I uptake was totally dependent on the presence of CO2. Type-II uptake consists usually of two uptake waves. Fluorescence transients measured in parallel give further support to the reality and interpretation of these two uptake waves. PA could thus provide a unique opportunity to monitor oxygen photoreduction in vivo with high sensitivity and time resolution.

Leaf chlorosis in oilseed rape plants (Brassica napus) grown on cadmium-polluted soil: causes and consequences for photosynthesis and growth.

Brassica napus L. (oilseed rape) was grown from seeds on a reconstituted soil contaminated with cadmium (100 mg Cd kg-1 dry soil), resulting in a marked chlorosis of the leaves which was investigated using a combination of biochemical, biophysical and physiological methods. Spectroscopic and chromatographic analyses of the photosynthetic pigments indicated that chlorosis was not due to a direct interaction of Cd with the chlorophyll biosynthesis pathway. In addition, mineral deficiency and oxidative stress were apparently not involved in the pigment loss. Leaf chlorosis was attributable to a marked decrease in the chloroplast density caused by a reduction in the number of chloroplasts per cell and a change in cell size, suggesting that Cd interfered with chloroplast replication and cell division. Relatively little Cd was found in the chloroplasts and the properties of the photosynthetic apparatus (electron transport, protein composition, chlorophyll antenna size, chloroplast ultrastructure) were not affected appreciably in plants grown on Cd-polluted soil. Depth profiling of photosynthetic pigments by phase-resolved photoacoustic spectroscopy revealed that the Cd-induced decrease in pigment content was very pronounced at the leaf surface (stomatal guard cells) compared to the leaf interior (mesophyll). This observation was consistent with light transmission and fluorescence microscopy analyses, which revealed that stomata density in the epidermis was noticeably reduced in Cd-exposed leaves. Concomitantly, the stomatal conductance estimated from gas-exchange measurements was strongly reduced with Cd. When plants were grown in a high-CO2 atmosphere (4,000 microliters CO2 l-1), the inhibitory effect of Cd on growth was not cancelled, suggesting that the reduced availability of CO2 at the chloroplast level associated with the low stomatal conductance was not the main component of Cd toxicity in oilseed rape.

Photodamage of the photosynthetic apparatus and its dependence on the leaf developmental stage in the npq1 Arabidopsis mutant deficient in the xanthophyll cycle enzyme violaxanthin de-epoxidase.

The npq1 Arabidopsis mutant is deficient in the violaxanthin de-epoxidase enzyme that converts violaxanthin to zeaxanthin in excess light (xanthophyll cycle). We have compared the behavior of mature leaves (ML) and developing leaves of the mutant and the wild type in various light environments. Thermoluminescence measurements indicated that high photon flux densities (>500 micromol m(-2) s(-1)) promoted oxidative stress in the chloroplasts of npq1 ML, which was associated with a loss of chlorophyll and an inhibition of the photochemical activity. Illuminating leaf discs in the presence of eosin, a generator of singlet oxygen, brought about pronounced lipid peroxidation in npq1 ML but not in wild-type leaves. No such effects were seen in young leaves (YL) of npq1, which were quite tolerant to strong light and eosin-induced singlet oxygen. Non-photochemical energy quenching was strongly inhibited in npq1 YL and ML and was not improved with high-light acclimation. Our results confirm that the xanthophyll cycle protects chloroplasts from photooxidation by a mechanism distinct from non-photochemical energy quenching and they reveal that the absence of xanthophyll cycle can be compensated by other protective mechanisms. npq1 YL were observed to accumulate considerable amounts of vitamin E during photoacclimation, suggesting that this lipophilic antioxidant could be involved in the high phototolerance of those leaves.

Salt shock-inducible photosystem I cyclic electron transfer in Synechocystis PCC6803 relies on binding of ferredoxin:NADP(+) reductase to the thylakoid membranes via its CpcD phycobilisome-linker homologous N-terminal domain.

Relative to ferredoxin:NADP(+) reductase (FNR) from chloroplasts, the comparable enzyme in cyanobacteria contains an additional 9 kDa domain at its amino-terminus. The domain is homologous to the phycocyanin associated linker polypeptide CpcD of the light harvesting phycobilisome antennae. The phenotypic consequences of the genetic removal of this domain from the petH gene, which encodes FNR, have been studied in Synechocystis PCC 6803. The in frame deletion of 75 residues at the amino-terminus, rendered chloroplast length FNR enzyme with normal functionality in linear photosynthetic electron transfer. Salt shock correlated with increased abundance of petH mRNA in the wild-type and mutant alike. The truncation stopped salt stress-inducible increase of Photosystem I-dependent cyclic electron flow. Both photoacoustic determination of the storage of energy from Photosystem I specific far-red light, and the re-reduction kinetics of P700(+), suggest lack of function of the truncated FNR in the plastoquinone-cytochrome b(6)f complex reductase step of the PS I-dependent cyclic electron transfer chain. Independent gold-immunodecoration studies and analysis of FNR distribution through activity staining after native polyacrylamide gelelectrophoresis showed that association of FNR with the thylakoid membranes of Synechocystis PCC 6803 requires the presence of the extended amino-terminal domain of the enzyme. The truncated DeltapetH gene was also transformed into a NAD(P)H dehydrogenase (NDH1) deficient mutant of Synechocystis PCC 6803 (strain M55) (T. Ogawa, Proc. Natl. Acad. Sci. USA 88 (1991) 4275-4279). Phenotypic characterisation of the double mutant supported our conclusion that both the NAD(P)H dehydrogenase complex and FNR contribute independently to the quinone cytochrome b(6)f reductase step in PS I-dependent cyclic electron transfer. The distribution, binding properties and function of FNR in the model cyanobacterium Synechocystis PCC 6803 will be discussed.

The violaxanthin cycle protects plants from photooxidative damage by more than one mechanism.

When light energy absorbed by plants becomes excessive relative to the capacity of photosynthesis, the xanthophyll violaxanthin is reversibly deepoxidized to zeaxanthin (violaxanthin cycle). The protective function of this phenomenon was investigated in a mutant of Arabidopsis thaliana, npq1, that has no functional violaxanthin deepoxidase. Two major consequences of the npq1 mutation are the absence of zeaxanthin formation in strong light and the partial inhibition of the quenching of singlet excited chlorophylls in the photosystem II light-harvesting complexes. Prolonged exposure of whole plants to bright light resulted in a limited photoinhibition of photosystem II in both npq1 and wild-type leaves, although CO(2) fixation and the linear electron transport in npq1 plants were reduced substantially. Lipid peroxidation was more pronounced in npq1 compared with the wild type, as measured by chlorophyll thermoluminescence, ethane production, and the total hydroperoxy fatty acids content. Lipid peroxidation was amplified markedly under chilling stress, and photooxidative damage ultimately resulted in leaf bleaching and tissue necrosis in npq1. The npq4 mutant, which possesses a normal violaxanthin cycle but has a limited capacity of quenching singlet excited chlorophylls, was rather tolerant to lipid peroxidation. The double mutant, npq4 npq1, which differs from npq4 only by the absence of the violaxanthin cycle, exhibited an increased susceptibility to photooxidative damage, similar to that of npq1. Our results demonstrate that the violaxanthin cycle specifically protects thylakoid membrane lipids against photooxidation. Part of this protection involves a mechanism other than quenching of singlet excited chlorophylls.

A chromodomain protein encoded by the arabidopsis CAO gene is a plant-specific component of the chloroplast signal recognition particle pathway that is involved in LHCP targeting.

A recessive mutation in Arabidopsis, named chaos (for chlorophyll a/b binding protein harvesting-organelle specific; designated gene symbol CAO), was isolated by using transposon tagging. Characterization of the phenotype of the chaos mutant revealed a specific reduction of pigment binding antenna proteins in the thylakoid membrane. These nuclear-encoded proteins utilize a chloroplast signal recognition particle (cpSRP) system to reach the thylakoid membrane. Both prokaryotes and eukaryotes possess a cytoplasmic SRP containing a 54-kD protein (SRP54) and an RNA. In chloroplasts, the homolog of SRP54 was found to bind a 43-kD protein (cpSRP43) rather than to an RNA. We cloned the CAO gene, which encodes a protein identified as Arabidopsis cpSRP43. The product of the CAO gene does not resemble any protein in the databases, although it contains motifs that are known to mediate protein-protein interactions. These motifs include ankyrin repeats and chromodomains. Therefore, CAO encodes an SRP component that is unique to plants. Surprisingly, the phenotype of the cpSRP43 mutant (i.e., chaos) differs from that of the Arabidopsis cpSRP54 mutant, suggesting that the functions of the two proteins do not strictly overlap. This difference also suggests that the function of cpSRP43 is most likely restricted to protein targeting into the thylakoid membrane, whereas cpSRP54 may be involved in an additional process(es), such as chloroplast biogenesis, perhaps through chloroplast-ribosomal association with chloroplast ribosomes.

Differential control of xanthophylls and light-induced stress proteins, as opposed to light-harvesting chlorophyll a/b proteins, during photosynthetic acclimation of barley leaves to light irradiance

Barley (Hordeum vulgare L.) plants were grown at different photon flux densities ranging from 100 to 1800 &mgr;mol m-2 s-1 in air and/or in atmospheres with reduced levels of O2 and CO2. Low O2 and CO2 partial pressures allowed plants to grow under high photosystem II (PSII) excitation pressure, estimated in vivo by chlorophyll fluorescence measurements, at moderate photon flux densities. The xanthophyll-cycle pigments, the early light-inducible proteins, and their mRNA accumulated with increasing PSII excitation pressure irrespective of the way high excitation pressure was obtained (high-light irradiance or decreased CO2 and O2 availability). These findings indicate that the reduction state of electron transport chain components could be involved in light sensing for the regulation of nuclear-encoded chloroplast gene expression. In contrast, no correlation was found between the reduction state of PSII and various indicators of the PSII light-harvesting system, such as the chlorophyll a-to-b ratio, the abundance of the major pigment-protein complex of PSII (LHCII), the mRNA level of LHCII, the light-saturation curve of O2 evolution, and the induced chlorophyll-fluorescence rise. We conclude that the chlorophyll antenna size of PSII is not governed by the redox state of PSII in higher plants and, consequently, regulation of early light-inducible protein synthesis is different from that of LHCII.

Reduction of the plastoquinone pool by exogenous NADH and NADPH in higher plant chloroplasts. Characterization of a NAD(P)H-plastoquinone oxidoreductase activity

Chlorophyll fluorescence measurements were performed on osmotically lysed potato chloroplasts in order to characterize the reactions involved in the dark reduction of photosynthetic inter-system chain electron carriers. Addition of NADH or NADPH to lysed chloroplasts increased the chlorophyll fluorescence level measured in the presence of a non-actinic light until reaching Fmax, thus indicating an increase in the redox state of the plastoquinone (PQ) pool. The fluorescence increase was more pronounced when the experiment was carried out under anaerobic conditions and was about 50% higher when NADH rather than NADPH was used as an electron donor. The NAD(P)H-PQ oxidoreductase reaction was inhibited by diphenylene iodonium, N-ethylmaleimide and dicoumarol, but insensitive to rotenone, antimycin A and piericidin A. By comparing the substrate specificity and the inhibitor sensitivity of this reaction to the properties of spinach ferredoxin-NADP+-reductase (FNR), we infer that FNR is not involved in the NAD(P)H-PQ oxidoreductase activity and conclude to the participation of rotenone-insensitive NAD(P)H-PQ oxidoreductase. By measuring light-dependent oxygen uptake in the presence of DCMU, methyl viologen and NADH or NADPH as an electron donors, the electron flow rate through the NAD(P)H-PQ oxidoreductase is estimated to about 160 nmol O2 min-1 mg-1 chlorophyll. The nature of this enzyme is discussed in relation to the existence of a thylakoidal NADH dehydrogenase complex encoded by plastidial ndh genes. Copyright 1998 Elsevier Science B.V.

Photoacoustically monitored thermal energy dissipation and xanthophyll cycle carotenoids in higher plant leaves.

When barley leaves were suddenly exposed to strong white light, the rate of thermal de-excitation of the absorbed light energy, measured with a photoacoustic device, increased by around 12% within a few minutes. This phenomenon was paralleled by a quenching of chlorophyll fluorescence. Simultaneous measurements of the heat emission increase and chlorophyll fluorescence quenching allowed the absolute yield phi F of chlorophyll fluorescence to be determined in vivo; phi F varied from around 2% (at Fo) to around 15% (at Fm) in a variety of plant species. No correlation was found between the time course of heat emission increase and the time course of violaxanthin-to-zeaxanthin conversion in barley leaves exposed to various light and temperature treatments, indicating that the zeaxanthin pool built up in the light was not directly involved in the increased heat emission. However, the operation of the violaxanthin cycle accelerated the photoinduced rise in thermal energy dissipation. Photoacoustic measurements on leaves of a zeaxanthin-accumulating Arabidopsis thaliana mutant lacking the violaxanthin cycle indicated that this acceleration could be ascribed to the disappearance of violaxanthin rather than to the formation of zeaxanthin.

Thermostability and Photostability of Photosystem II in Leaves of the Chlorina-f2 Barley Mutant Deficient in Light-Harvesting Chlorophyll a/b Protein Complexes.

The chlorophyll-b-less chlorina-f2 barley mutant is deficient in the major as well as some minor light-harvesting chlorophyll-protein complexes of photosystem II (LHCII). Although the LHCII deficiency had relatively minor repercussions on the leaf photosynthetic performances, the responses of photosystem II (PSII) to elevated temperatures and to bright light were markedly modified. The chlorina-f2 mutation noticeably reduced the thermostability of PSII, with thermal denaturation of PSII starting at about 35[deg]C and 38.5[deg]C in chlorina-f2 and in the wild type, respectively. The increased susceptibility of PSII to heat stress in chlorina-f2 leaves was due to the weakness of its electron donor side, with moderate heat stress causing detachment of the 33-kD extrinsic PSII protein from the oxygen-evolving complex. Prolonged dark adaptation of chlorina-f2 leaves was also observed to inhibit the PSII donor side. However, weak illumination slowly reversed the dark-induced inhibition of PSII in chlorina-f2 and cancelled the difference in PSII thermostability observed between chlorina-f2 and wild-type leaves. The mutant was more sensitive to photoinhibition than the wild type, with strong light stress impairing the PSII donor side in chlorina-f2 but not in the wild type. This difference was not observed in anaerobiosis or in the presence of 3-(3,4-dichlorophenyl)- 1,1-dimethylurea, diuron. The acceptor side of PSII was only slightly affected by the mutation and/or the aforementioned stress conditions. Taken together, our results indicate that LHCII stabilize the PSII complexes and maintain the water-oxidizing system in a functional state under varying environmental conditions.

Thylakoid membrane fluidity and thermostability during the operation of the xanthophyll cycle in higher-plant chloroplasts.

Barley leaves were exposed for several min to a white light of photon flux density 1000 micromol m-2 s-1, leading to a massive conversion of the xanthophyll violaxanthin to antheraxanthin and zeaxanthin in the absence of lipid peroxidation. Using electron spin resonance spectroscopy and different spin-labeled stearate probes, we observed that this light treatment noticeably decreased thylakoid membrane lipid fluidity. The light-induced membrane rigidification (i) was proportional to the amount of zeaxanthin present in the membranes, (ii) was blocked by dithiothreitol, a potent inhibitor of the violaxanthin de-epoxidase, (iii) was slowly reversible in the dark, (iv) was not observed in thylakoids of an Arabidopsis mutant that has no xanthophyll cycle and (v) was accompanied by a substantial increase in the thermostability of the ionic permeability properties of the thylakoid membranes. The amount of xanthophyll-cycle pigments found in photosystem II was observed to significantly decrease after illumination. Photoacoustic and chlorophyll fluorometric analyses of the illuminated leaves revealed that strong illumination decreased the quantum yield of photosynthetic oxygen evolution and the pigment antenna size of photosystem II in green light (preferentially absorbed by carotenoids) but not in red light (absorbed by chlorophylls only). Taken together in the light of previous in vitro data on carotenoids incorporated into artificial membranes, our results indicate that the xanthophyll cycle could be an 'emergency mechanism' that rapidly provides thylakoid membrane lipids with rigidifying carotenoid molecules upon sudden increase in light intensity. The significance of this mechanism for the membrane function and adaptation to stressful light and temperature conditions is discussed.

Photosynthesis, chlorophyll fluorescence, light-harvesting system and photoinhibition resistance of a zeaxanthin-accumulating mutant of Arabidopsis thaliana.

The abscisic-acid-deficient aba-1 mutant of Arabidopsis thaliana is unable to epoxidize zeaxanthin. As a consequence, it contains large amounts of this carotenoid and lacks epoxy-xanthophylls. HPLC analysis of pigment contents in leaves, isolated thylakoids and preparations of the major light-harvesting complex of photosystem II (PSII) (LHC-II) indicated that zeaxanthin replaced neoxanthin, violaxanthin and antheraxanthin in the light-harvesting system of PSII in aba-1. Non-denaturing electrophoretic fractionation of solubilized thylakoids showed that the xanthophyll imbalance in aba-1 was associated with a pronounced decrease in trimeric LHC-II in favour of monomeric complexes, with a substantial increase in free pigments (mainly zeaxanthin and chlorophyll b), suggesting a decreased stability of LHC-II. The reduced thermostability of PSII in aba-1 was also deduced from in vivo chlorophyll fluorescence measurements. Wild-type and aba-1 leaves could not be distinguished on the basis of their photosynthetic performance: no significant difference was observed between the two types of leaves for light-limited and light-saturated photosynthetic oxygen evolution, PSII photochemistry and PSII to PSI electron flow. When dark-adapted leaves (grown in white light of 80 mumol m-2s-1) were suddenly exposed to red light of 150 mumol m-2s-1, there was a strong nonphotochemical quenching of chlorophyll fluorescence, the amplitude of which was virtually identical (at steady state) in aba-1 and wild-type leaves, despite the fact that the xanthophyll cycle pigment pool was completely in the form of zeaxanthin in aba-1 and almost exclusively in the form of violaxanthin in the wild type. A high concentration of zeaxanthin in aba-1 thylakoids did not, in itself, provide any particular protection against the photoinhibition of PSII. Taken together, the presented results indicate the following: (1) zeaxanthin can replace epoxy-xanthophylls in LHC-II without significantly affecting the photochemical efficiency of PSII; (2) zeaxanthin does not play any specific role in direct (thermal) energy dissipation in PSII; (3) the photoprotective action of the xanthophyll cycle (rapid photoconversion of violaxanthin to zeaxanthin) is not based on the mere substitution of violaxanthin by zeaxanthin in the chlorophyll antennae.

Short-term responses of Photosystem I to heat stress : Induction of a PS II-independent electron transport through PS I fed by stromal components.

When 23°C-grown potato leaves (Solanum tuberosum L.) were exposed for 15 min to elevated temperatures in weak light, a dramatic and preferential inactivation of Photosystem (PS) II was observed at temperatures higher than about 38°C. In vivo photoacoustic measurements indicated that, concomitantly with the loss of PS II activity, heat stress induced a marked gas-uptake activity both in far-red light (>715 nm) exciting only PS I and in broadband light (350-600 nm) exciting PS I and PS II. In view of its suppression by nitrogen gas and oxygen and its stimulation by high carbon-dioxide concentrations, the bulk of the photoacoustically measured gas uptake by heat-stressed leaves was ascribed to rapid carbon-dioxide solubilization in response to light-modulated stroma alkalization coupled to PS I-driven electron transport. Heat-induced gas uptake was observed to be insensitive to the PS II inhibitor diuron, sensitive to the plastocyanin inhibitor HgCl2 and saturated at a rather high photon flux density of around 1200 µE m(-2) s(-1). Upon transition from far-red light to darkness, the oxidized reaction center P700(+) of PS I was re-reduced very slowly in control leaves (with a half time t1/2 higher than 500 ms), as measured by leaf absorbance changes at around 820 nm. Heat stress caused a spectacular acceleration of the postillumination P700(+) reduction, with t1/2 falling to a value lower than 50 ms (after leaf exposure to 48°C). The decreased t1/2 was sensitive to HgCl2 and insensitive to diuron, methyl viologen (an electron acceptor of PS I competing with the endogenous acceptor ferredoxin) and anaerobiosis. This acceleration of the P700(+) reduction was very rapidly induced by heat treatment (within less than 5 min) and persisted even after prolonged irradiation of the leaves with far-red light. After heat stress, the plastoquinone pool exhibited reduction in darkness as indicated by the increase in the apparent Fo level of chlorophyll fluorescence which could be quenched by far-red light. Application (for 1 min) of far-red light to heat-pretreated leaves also induced a reversible quenching of the maximal fluorescence level Fm, suggesting formation of a pH gradient in far-red light. Taken together, the presented data indicate that PS I responded to the heat-induced loss of PS II photochemical activity by catalyzing an electron flow from stromal reductants. Heat-stress-induced PS I electron transport independent of PS II seems to constitute a protective mechanism since block of this electron pathway in anaerobiosis was observed to result in a dramatic photoinactivation of PS I.

The protective function of the xanthophyll cycle in photosynthesis.

The rapid conversion of the carotenoid violaxanthin to zeaxanthin via antheraxanthin (xanthophyll cycle) in potato leaves exposed at 23 degrees C to a strong white light of 2000 microE.m-2.s-1 was associated with a slight inhibition of photosynthetic electron transport (as estimated from chlorophyll fluorescence measurements) and a low lipid peroxidation (as estimated from ethane measurements). When the xanthophyll cycle was blocked by dithiothreitol (3 mM) or low temperature (3 degrees C), photoinhibition of electron transport was exacerbated and pronounced lipid peroxidation occurred concomitantly. Accumulation of zeaxanthin and antheraxanthin in potato leaves by a non-photoinhibitory light treatment at 23 degrees C (900 microE.m-2.s-1 for 1 h) considerably reduced the level of lipid peroxidation during subsequent light stress at 3 degrees C. The presented results indicate that one of the functions of the xanthophyll cycle could be the protection of thylakoid membranes against lipid peroxidation, suggesting that zeaxanthin and antheraxanthin synthesized in strong light are present as free pigments in the membrane lipid bilayer.

Photoinhibition of photosynthesis in chilled potato leaves is not correlated with a loss of Photosystem-II activity : Preferential inactivation of Photosystem I.

When 23 °C-grown potato leaves (Solanum tuberosum L.) were irradiated at 23 °C with a strong white light, photosynthetic electron transport and Photosystem-II (PS II) activity were inhibited in parallel. When the light treatment was given at a low temperature of 3 °C, the photoinhibition of photosynthesis was considerably enhanced, as expected. Surprisingly, no such stimulation of photoinhibition was observed with respect to the PS II function. A detailed functional analysis of the photosynthetic apparatus, using in-vivo fluorescence, absorbance, oxygen and photoacoustic measurements, and artificial electron donors/acceptors, showed a pronounced alteration of PS I activity during light stress at low temperature. More precisely, it was observed that both the pool of photooxidizeable reaction center pigment (P700) of PS I and the efficiency of PS I to oxidize P700 were dramatically reduced. Loss of P700 activity was shown to be essentially dependent on atmospheric O2 and to require a continued flow of electrons from PS II, suggesting the involvement of the superoxide anion radical which is produced by the interaction of O2 and the photosynthetic electron-transfer chain through the Mehler reaction. Mass spectrometric measurements of O2 exchange by potato leaves under strong illumination did not reveal, however, any stimulation of the Mehler reaction at low temperature, thus leading to the conclusion that O2 toxicity mainly resulted from a chilling-induced inhibition of the scavenging system for O2-radicals. Support for this interpretation was provided by the light response of potato leaves infiltrated with an inhibitor (diethyldithiocarbamate) of the chloroplastic Cu-Zn superoxide dismutase. It was indeed possible to simulate the differential inhibition of the PS II photochemical activity and the linear electron transport observed during light stress at low temperature by illuminating at 23 °C diethyldithiocarbamate-poisoned leaves. The experimental data presented here suggests that (i) the previously reported resistance of PS I to photoinhibition damage in-vivo is not an intrinsic property of PS I but results from efficient protective systems against O2 toxicity, (ii) PS I is photoinhibited in chilled potato leaf due to the inactivation of this PS I defence system and (iii) PS I is more sensitive to superoxide anion radicals than PS II.

Stress Tolerance of Photosystem II in Vivo: Antagonistic Effects of Water, Heat, and Photoinhibition Stresses.

The in vivo photochemical activity of photosystem II was inferred from modulated chlorophyll fluorescence and photoacoustic measurements in intact leaves of several plant species (Lycopersicon esculentum Mill., Solanum tuberosum L., Solanum nigrum L.) exposed to various environmental stresses (drought, heat, strong light) applied separately or in combination. Photosystem II was shown to be highly drought-resistant: even a drastic desiccation in air of detached leaf samples only marginally affected the quantum yield for photochemistry in photosystem II. However, water stress markedly modified the responses of photosystem II to superimposed constraints. The stability of photosystem II to heat was observed to increase strongly in leaves exposed to water stress conditions: heat treatments (e.g. 42 degrees C in the dark), which caused a complete and irreversible inhibition of photosystem II in well-watered (tomato) leaves, resulted in a small and fully reversible reduction of the photochemical efficiency of photosystem II in drought-stressed leaves. In vivo photoacoustic data indicated that photosystem I was highly resistant to both heat and water stresses. When leaves were illuminated with intense white light at 25 degrees C, photoinhibition damage of photosystem II was more pronounced in water-stressed leaves than in undesiccated controls. However, in nondehydrated leaves, photoinhibition of photosystem II was strongly temperature dependent, being drastically stimulated at high temperatures above 38 to 40 degrees C. As a consequence, when exposed to strong light at high temperature, photosystem II photochemistry was significantly less inhibited in dehydrated leaves than in control well-hydrated leaves. Our results demonstrate the existence of a marked antagonism between physicochemical stresses, with water stress enhancing the resistance of photosystem II to constraints (heat, strong light at high temperature) that are usually associated with drought in the field.

Dynamics of electron transfer within and between PS II reaction center complexes indicated by the light-saturation curve of in vivo variable chlorophyll fluorescence emission.

The dynamics of light-induced closure of the PS II reaction centers was studied in intact, dark-adapted leaves by measuring the light-irradiance (I) dependence of the relative variable chlorophyll fluorescence V which is the ratio between the amplitude of the variable fluorescence induced by a pulse of actinic light and the maximal variable fluorescence amplitude obtained with an intense, supersaturating light pulse. It is shown that the light-saturation curve of V is a hyperbola of order n. The experimental values of n ranged from around 0.75 to around 2, depending on the plant material and the environmental conditions. A simple theoretical analysis confirmed this hyperbolic relationship between V and I and suggested that n could represent the apparent number of photons necessary to close one reaction center. Thus, experimental conditions leading to n values higher than 1 could indicate that, from a macroscopic viewpoint, more than one photon is necessary to close one PS II center, possibly due to changes in the relative concentrations of the different redox states of the PS II reaction center complexes at the quasi-steady state induced by the actinic light. On the other hand, the existence of environmental conditions resulting in n noticeably lower than 1 suggests the possibility of an electron flow between PS II reaction center complexes.

[Peridural obstetrical analgesia and the cicatricial uterus. A 4-year evaluation]. / Analgésie péridurale obstétricale et utérus cicatriciel. Bilan de quatre années.

191 women with previous cesarean section in 291 indexed between October 1985 and September 1989 underwent a trial of labor. 146 patients received epidural analgesia in the course of labor. Vaginal delivery occurred in 126 patients (86.3%). Duration for epidural analgesia in labour was 163 +/- 110 min. The intrauterine pressure for monitoring continuous was 51.00 +/- 15.40 mmHg. We report four uterine dehiscences and one rupture. In no case, epidural analgesia did not delay the diagnosis. The use of epidural analgesia for trial of labor in previous cesarean section did not increase maternal or fetal risk.