Effect of green light on the amount and activity of NDH-1–PSI supercomplex in Synechocystis sp. strain PCC 6803

Photosynthetica - Cyanobacterial NDH-1 interacts with PSI to form NDH-1–PSI supercomplex. CpcG2, a linker protein for the PSI-specific peripheral antenna CpcG2-phycobilisome, is essential for...     

Sensitivity of the photosynthetic apparatus to UV-A radiation: role of light-harvesting complex II–photosystem II supercomplex organization

In this work we study the effect of UV-A radiation on the function of the photosynthetic apparatus in thylakoid membranes with different organization of the light-harvesting complex II–photosystem II (LHCII–PSII) supercomplex. Leaves and isolated thylakoid membranes from a number of previously characterized pea species with different LHCII size and organization were subjected to UV-A treatment. A relationship was found between the molecular organization of the LHCII (ratio of the oligomeric to monomeric forms of LHCII) and UV-A-induced changes both in the energy transfer from PSII to PSI and between the chlorophyll–protein complexes within the LHCII–PSII supercomplex. Dependence on the organization of the LHCII was also found with regard to the degree of inhibition of the photosynthetic oxygen evolution. The susceptibility of energy transfer and oxygen evolution to UV-A radiation decreased with increasing LHCII oligomerization when the UV-A treatment was performed on isolated thylakoid membranes, in contrast to the effect observed in thylakoid membranes isolated from pre-irradiated pea leaves. The data suggest that UV-A radiation leads mainly to damage of the PSIIα centers. Comparison of membranes with different organization of their LHCII–PSII supercomplex shows that the oligomeric forms of LHCII play a key role for sensitivity to UV-A radiation of the photosynthetic apparatus. S. G. Taneva is Associated member of the Institute of Biophysics, Bulgarian Academy of Sciences.     

Photosynthetic acclimation: does the dynamic structure and macro-organisation of photosystem II in higher plant grana membranes regulate light harvesting states?

The efficiency of light harvesting in higher plant photosynthesis is regulated in response to external environmental conditions. Under conditions of excess light, the normally highly efficient light-harvesting system of photosystem II is switched into a state in which unwanted, potentially harmful, energy is dissipated as heat. This process, known as nonphotochemical quenching, occurs by the creation of energy quenchers following conformational change in the light-harvesting complexes, which is initiated by the build up of the thylakoid pH gradient and controlled by the xanthophyll cycle. In the present study, the evidence to support the notion that this regulatory mechanism is dependent upon the organization of the different antenna subunits in the stacked grana membranes is reviewed. We postulate that nonphotochemical quenching occurs within a structural locus comprising the PsbS subunit and components of the light-harvesting antenna, CP26, CP24, CP29 and LHCIIb (the major trimeric light-harvesting complex), formed in response to protonation and controlled by the xanthophyll cycle carotenoids.     

Formation of superoxide anion and carbon-centered radicals by photosystem II under high light and heat stress—EPR spin-trapping study

In this study, electron paramagnetic resonance spin-trapping spectroscopy was used to study the light-induced production of superoxide anion (O₂ ^?-) and carbon-centered (R^?) radicals by Photosystem II (PSII). It is evidenced here that exposure of PSII membranes to high light (2,000 μmol photons m^?2 s^?1) or heat (47 °C) treatments prior to the illumination suppressed O₂ ^?- production, while R^? was formed. Formation of R^? in the both high light- and heat-treated PSII membranes was enhanced by DCMU. Removal of molecular oxygen by glucose/glucose oxidase/catalase system and O₂ ^?- scavenging by exogenous superoxide dismutase completely suppressed carbon-centered radical formation. It is proposed here that the oxidation of polyunsaturated fatty acids and amino acids by O₂ ^?- on the electron acceptor side of PSII results in the formation of R^?, known to initiate a cascade reaction leading to the lipid peroxidation and protein degradation, respectively.     

Stability of the S₃and S₂state intermediates in photosystem II directly probed by EPR spectroscopy

The stability of the S(3) and S(2) states of the oxygen evolving complex in photosystem II (PSII) was directly probed by EPR spectroscopy in PSII membrane preparations from spinach in the presence of the exogenous electron acceptor PpBQ at 1, 10, and 20 °C. The decay of the S(3) state was followed in samples exposed to two flashes by measuring the split S(3) EPR signal induced by near-infrared illumination at 5 K. The decay of the S(2) state was followed in samples exposed to one flash by measuring the S(2) state multiline EPR signal. During the decay of the S(3) state, the S(2) state multiline EPR signal first increased and then decreased in amplitude. This shows that the decay of the S(3) state to the S(1) state occurs via the S(2) state. The decay of the S(3) state was biexponential with a fast kinetic phase with a few seconds decay half-time. This occurred in 10-20% of the PSII centers. The slow kinetic phase ranged from a decay half-time of 700 s (at 1 °C) to ~100 s (at 20 °C) in the remaining 80-90% of the centers. The decay of the S(2) state was also biphasic and showed quite similar kinetics to the decay of the S(3) state. Our experiments show that the auxiliary electron donor Y(D) was oxidized during the entire experiment. Thus, the reduced form of Y(D) does not participate to the fast decay of the S(2) and S(3) states we describe here. Instead, we suggest that the decay of the S(3) and S(2) states reflects electron transfer from the acceptor side of PSII to the donor side of PSII starting in the corresponding S state. It is proposed that this exists in equilibrium with Y(Z) according to S(3)Y(Z) ? S(2)Y(Z)(?) in the case of the S(3) state decay and S(2)Y(Z) ? S(1)Y(Z)(?) in the case of the S(2) state decay. Two kinetic models are discussed, both developed with the assumption that the slow decay of the S(3) and S(2) states occurs in PSII centers where Y(Z) is also a fast donor to P(680)(+) working in the nanosecond time regime and that the fast decay of the S(3) and S(2) states occurs in centers where Y(Z) reduces P(680)(+) with slower microsecond kinetics. Our measurements also demonstrate that the split S(3) EPR signal can be used as a direct probe to the S(3) state and that it can provide important information about the redox properties of the S(3) state.     

A new mechanism for adaptation to changes in light intensity and quality in the red alga,Porphyra perforata. II. Characteristics of state II—state III transitions

Characteristics of state II—state III transitions in the red alga, Porphyra perforata, were studied by measuring the fluorescence time course at room temperature and fluorescence spectra at 77 K. The state II to III transition was induced by system II light and was sensitive to uncouplers of photophosphorylation. This state II to III transition has a dark step(s) that could be easily separated from the light process. A state III to II transition occurred in the dark, but system I light accelerated the transition. The accelerating effect of system I light was not sensitive to uncouplers of photophosphorylation, but was inhibited by the addition of valinomycin + KCl or antimycin A. Compared to state I—state II transitions, the state II—state III transitions occurred more rapidly. The state II to state III transitions are different from the state I to state II transitions in that in state III the activity of photosystem II is changed without having any effect on photosystem I activity (Satoh and Fork, Biochim. Biophys, Acta, in press, 1982). It is suggested that the state II—state III transition represents a mechanism by which the alga can avoid photodamage resulting from absorption of excess light energy by photosystem II.     

Photochemical metabolism and fruit quality of Ubá mango tree exposed to combined light and heat stress in the field

Several experiments have highlighted the complexity of stress interactions, in field conditions, involved in plant response. However, these impacts on the mechanisms involved in plant photosynthetic response remains understudied. The aim of this work was to compare the photosynthetic efficiencies and fruit quality of mango tree (Mangifera indica L.) cv. Ubá harvested from plants cultivated on the east and west sides of a commercial orchard, according to the position of plants in relation to sunrise. Chlorophyll a fluorescence, was analyzed in leaves in four different periods: fruit growth phase, fruit ripening phase, post-harvest period and after plant pruning. Photoinhibitory damage was detected by the trapped energy flux and transported electron flux per reaction center during the fruit ripening phase, and by specific energy fluxes and yield quantum efficiency after plant pruning. Although high radiation caused photoinhibition on leaves from plants cultivated on the west side of the orchard, it provided sweeter fruits. In contrast to our initial hypothesis, it was verified that plants cultivated on the west side of the orchard presented better photochemical performance in periods with the greatest requirements of photoassimilates. In addition, plants demonstrated different abilities to deal with changes on photosynthetic active radiation and high temperature. This information suggests that the phenotypic plasticity of the Ubá mango cultivar is considerable, which can be exploited to be used in regions with great relief variations and the combination of increased irradiance and high temperature.     

Effect of protein aggregation on the spectroscopic properties and excited state kinetics of the LHCII pigment–protein complex from green plants

Steady-state and time-resolved absorption and fluorescence spectroscopic experiments have been carried out at room and cryogenic temperatures on aggregated and unaggregated monomeric and trimeric LHCII complexes isolated from spinach chloroplasts. Protein aggregation has been hypothesized to be one of the mechanistic factors controlling the dissipation of excess photo-excited state energy of chlorophyll during the process known as nonphotochemical quenching. The data obtained from the present experiments reveal the role of protein aggregation on the spectroscopic properties and dynamics of energy transfer and excited state deactivation of the protein-bound chlorophyll and carotenoid pigments.     

Decay kinetics and quantum yields of fluorescence in photosystem I from Synechococcus elongatus with P700 in the reduced and oxidized state: are the kinetics of excited state decay trap-limited or transfer-limited?

Transfer and trapping of excitation energy in photosystem I (PS I) trimers isolated from Synechococcus elongatus have been studied by an approach combining fluorescence induction experiments with picosecond time-resolved fluorescence measurements, both at room temperature (RT) and at low temperature (5 K). Special attention was paid to the influence of the oxidation state of the primary electron donor P700. A fluorescence induction effect has been observed, showing a approximately 12% increase in fluorescence quantum yield upon P700 oxidation at RT, whereas at temperatures below 160 K oxidation of P700 leads to a decrease in fluorescence quantum yield ( approximately 50% at 5 K). The fluorescence quantum yield for open PS I (with P700 reduced) at 5 K is increased by approximately 20-fold and that for closed PS I (with P700 oxidized) is increased by approximately 10-fold, as compared to RT. Picosecond fluorescence decay kinetics at RT reveal a difference in lifetime of the main decay component: 34 +/- 1 ps for open PS I and 37 +/- 1 ps for closed PS I. At 5 K the fluorescence yield is mainly associated with long-lived components (lifetimes of 401 ps and 1.5 ns in closed PS I and of 377 ps, 1.3 ns, and 4.1 ns in samples containing approximately 50% open and 50% closed PS I). The spectra associated with energy transfer and the steady-state emission spectra suggest that the excitation energy is not completely thermally equilibrated over the core-antenna-RC complex before being trapped. Structure-based modeling indicates that the so-called red antenna pigments (A708 and A720, i.e., those with absorption maxima at 708 nm and 720 nm, respectively) play a decisive role in the observed fluorescence kinetics. The A720 are preferentially located at the periphery of the PS I core-antenna-RC complex; the A708 must essentially connect the A720 to the reaction center. The excited-state decay kinetics turn out to be neither purely trap limited nor purely transfer (to the trap) limited, but seem to be rather balanced.     

Induced fit or conformational selection? The role of the semi-closed state in the maltose binding protein

A full characterization of the thermodynamic forces underlying ligand-associated conformational changes in proteins is essential for understanding and manipulating diverse biological processes, including transport, signaling, and enzymatic activity. Recent experiments on the maltose binding protein (MBP) have provided valuable data about the different conformational states implicated in the ligand recognition process; however, a complete picture of the accessible pathways and the associated changes in free energy remains elusive. Here we describe results from advanced accelerated molecular dynamics (aMD) simulations, coupled with adaptively biased force (ABF) and thermodynamic integration (TI) free energy methods. The combination of approaches allows us to track the ligand recognition process on the microsecond time scale and provides a detailed characterization of the protein's dynamic and the relative energy of stable states. We find that an induced-fit (IF) mechanism is most likely and that a mechanism involving both a conformational selection (CS) step and an IF step is also possible. The complete recognition process is best viewed as a "Pac Man" type action where the ligand is initially localized to one domain and naturally occurring hinge-bending vibrations in the protein are able to assist the recognition process by increasing the chances of a favorable encounter with side chains on the other domain, leading to a population shift. This interpretation is consistent with experiments and provides new insight into the complex recognition mechanism. The methods employed here are able to describe IF and CS effects and provide formally rigorous means of computing free energy changes. As such, they are superior to conventional MD and flexible docking alone and hold great promise for future development and applications to drug discovery.     

Phase transitions in nuclei and chromatin. Is nuclear volume controlled by the chromatin or by the nuclear matrix?

Changes in the volume of rat liver nuclei have been monitored as a function of modifications in ionic environment (from 0 to 20 mM), temperature (from 4 to 37 degrees C), and pH (from 1 to 8). An abrupt reduction of nuclear volume occurred with increasing ion concentration, this contraction being more pronounced with bivalent (either Ca2+ or Mg2+) than with monovalent (either Na+ or K+) cations. The lowering of pH produced a similar effect. Parallel changes in chromatin structure took place at the same time as phase-like transitions. Atomic absorption spectroscopy allowed determination of free and nuclei-bound ions, pointing to the presence of a sizeable number of free binding sites for chromatin-DNA even within intact nuclei. DNA-phosphate sites appear to be neutralized by ions strictly according to the size of the electric charge and polyelectrolyte theory. Partial digestion (by micrococcal nuclease) or simple breaks (by chemical carcinogens) of the chromatin-DNA fiber caused respectively elimination or reduction of the abrupt volume changes in the intact nuclei. The apparent role of chromatin structure versus nuclear matrix in determining the shape and volume of intact nuclei is briefly discussed.     

Nitrification rates of algal–bacterial biofilms in wastewater stabilization ponds under light and dark conditions

The objective of this study was to investigate nitrification rates in algal–bacterial biofilms of waste stabilization ponds (WSP) under different conditions of light, oxygen and pH. Biofilms were grown on wooden plates of 6.0 cm by 8.0 cm by 0.4 cm in a PVC tray continuously fed with synthetic wastewater with initial NH₄-N and Chemical Oxygen Demand (COD) concentrations of 40 mg l^?1 and 100 mg l^?1, respectively, under light intensity of 85–95 μE m^?2 s^?1. Batch activity tests were carried out by exposure of the plates to light conditions as above (to simulate day time), dim light of 1.8–2.2 μE m^?2 s^?1 (to simulate reduced light as in deeper locations in WSP) and dark conditions (to simulate night time). Dissolved oxygen (DO) concentration and pH were controlled. At some experiments, both parameters were kept constant, and at others they were left to vary as in WSP. Results show biofilm nitrification rates of 945–1817 mg-N m^?2 d^?1 and 1124–1615 mg-N m^?2 d^?1 for light and dark experiments. When the minimum DO was 4.1 mg l^?1, the biofilm nitrification rates under light and dark conditions did not differ significantly at 95% confidence. When the minimum DO in the dim light experiment was 3.2 mg l^?1, the nitrification rates under light and dim light conditions were 945 mg-N m^?2 d^?1 and 563 mg-N m^?2 d^?1 and these significantly differed. Further decrease of DO to 1.1 mg l^?1 under dark conditions resulted in more decrease of the nitrification rates to 156 mg-N m^?2 d^?1. It therefore seems that under these experimental conditions, biofilm nitrification rates are significantly reduced at a certain point when bulk water DO is between 3.2 mg l^?1 and 4.1 mg l^?1. As long as bulk water DO under dark is high, light is not important in influencing the process of nitrification.     

Study of the activity of quaternary ammonium compounds in the mitigation of biofouling in heat exchangers–condensers cooled by seawater

The effectiveness of two quaternary ammonium compounds (QACs) (non-oxidising biocides) to reduce the growth of biofilm adhering to the tubes of a heat exchanger-condenser cooled by seawater was evaluated. Their effectiveness was compared to that of a conventional oxidising biocide (sodium hypochlorite [NaOCl]) under the same testing conditions. Each biocide was applied intermittently (6?h on, 6?h off) in a first shock stage (1.5?ppm over 8?days) and a second stabilising stage (0.5?ppm over 20?days). The results showed that the antifouling effectiveness of the first of the QACs (fifth generation) was comparable to that shown by the oxidising power of NaOCl. Although the reaction time was longer than that of NaOCl, both the compounds removed the biofilm, and the tube was practically restored to its clean condition. Treatment with the second of the QACs (fourth generation) allowed for the stabilisation of biofilm growth, but not for its removal. Ecotoxicology studies classified the QACs as environmentally harmless under the testing conditions.     

The site of regulation of light capture in Symbiodinium: Does the peridinin–chlorophyll a–protein detach to regulate light capture?

Dinoflagellates from the genus Symbiodinium form symbiotic associations with cnidarians including corals and anemones. The photosynthetic apparatuses of these dinoflagellates possess a unique photosynthetic antenna system incorporating the peridinin–chlorophyll a–protein (PCP). It has been proposed that the appearance of a PCP-specific 77 K fluorescence emission band around 672–675 nm indicates that high light treatment results in PCP dissociation from intrinsic membrane antenna complexes, blocking excitation transfer to the intrinsic membrane-bound antenna complexes, chlorophyll a–chlorophyll c₂–peridinin–protein-complex (acpPC) and associated photosystems (Reynolds et al., 2008 Proc Natl Acad Sci USA 105:13674–13678).We have tested this model using time-resolved fluorescence decay kinetics in conjunction with global fitting to compare the time-evolution of the PCP spectral bands before and after high light exposure. Our results show that no long-lived PCP fluorescence emission components appear either before or after high light treatment, indicating that the efficiency of excitation transfer from PCP to membrane antenna systems remains efficient and rapid even after exposure to high light. The apparent increased relative emission at around 675 nm was, instead, caused by strong preferential exciton quenching of the membrane antenna complexes associated with acpPC and reaction centers. This strong non-photochemical quenching (NPQ) is consistent with the activation of xanthophyll-associated quenching mechanisms and the generally-observed avoidance in nature of long-lived photoexcited states that can lead to oxidative damage. The acpPC component appears to be the most strongly quenched under high light exposure suggesting that it houses the photoprotective exciton quencher.     

Thermal acclimation,heat shock and photoperiod: Do these factors interplay in the adaptive responses of crab neuromuscular systems to temperature?

Evidence is reviewed demonstrating that the adaptive responses to temperature made by the walking leg neuromuscular system of crabs (Carcinus maenas) are in response to local temperature and not in response to hierarchical influences by the CNS and hormonal systems.     

Determination of glassy state by cryo-SEM and DSC in cryopreservation of mint shoot tips by encapsulation–dehydration

Cryopreservation of mint shoot tips grown in vitro (Mentha × piperita) was performed after encapsulation in alginate beads. Encapsulated shoot tips were first precultured in sucrose enriched medium (0.75 M) and then dried under a sterile air flow (0–6 h). After cooling in liquid nitrogen and warming in a warm water bath, alginate beads were transferred to solid culture medium for 4 weeks. The effect of dehydration time of the encapsulated shoots was evaluated for water content, cooling and warming rates, ice crystal formation and cellular vitrification, by using low temperature scanning electron microscopy and differential scanning calorimetry. Viability of the recovered material showed a close relation between the dehydration time, cooling and warming rates, ice formation avoidance and tissue vitrification. At short drying periods (up to 3 h), ice crystals were formed and the viability was low or absent. After longer drying periods (5 and 6 h), both beads and specimens became vitrified.     

Estimation of relative contribution of “mobile phycobilisome” and “energy spillover” in the light–dark induced state transition in Spirulina platensis

Previously, it was clarified that phycobilisome (PBS) mobility and energy spillover were both involved in light-to-dark induced state transitions of intact Spirulina platensis cells. In this work, by taking advantage of the characteristic fluorescence spectra of photosystem I (PSI) trimers and monomers as indicators, the relative contributions for the “mobile PBS” and “energy spillover” are quantitatively estimated by separating the fluorescence contribution of PBS mobility from that of PSI oligomeric change. Above the phase transition temperature (T _PT) of the membrane lipids, the relative proportion of the contributions is invariable with 65% of “mobile PBS” and 35% of “energy spillover”. Below T _PT, the proportion for the “mobile PBS” becomes larger under lowering temperature even reaching 95% with 5% “energy spillover” at 0°C. It is known that lower temperature leads to a further light state due to a more reduced or oxidized PQ pool. Based on the current result, it can be deduced that disequilibrium of the redox state of the PQ pool will trigger PBS movement instead of change in the PSI oligomeric state.     

Adjuncts or adversaries to shared decision-making? Applying the Integrative Model of behavior to the role and design of decision support interventions in healthcare interactions

Background

Clinical practice guidelines have been a popular tool for the improvement of health care through the implementation of evidence from systematic research. Yet, it is increasingly clear that knowledge alone is insufficient to change practice. The social, cultural, and material contexts within which practice occurs may invite or reject innovation, complement or inhibit the activities required for success, and sustain or alter adherence to entrenched practices. However, knowledge translation (KT) models are limited in providing insight about how and why contextual contingencies interact, the causal mechanisms linking structural aspects of context and individual agency, and how these mechanisms influence KT. Another limitation of KT models is the neglect of methods to engage potential adopters of the innovation in critical reflection about aspects of context that influence practice, the relevance and meaning of innovation in the context of practice, and the identification of strategies for bringing about meaningful change.

Discussion

This paper presents a KT model, the Critical Realism and the Arts Research Utilization Model (CRARUM), that combines critical realism and arts-based methodologies. Critical realism facilitates understanding of clinical settings by providing insight into the interrelationship between its structures and potentials, and individual action. The arts nurture empathy, and can foster reflection on the ways in which contextual factors influence and shape clinical practice, and how they may facilitate or impede change. The combination of critical realism and the arts within the CRARUM model promotes the successful embedding of interventions, and greater impact and sustainability.

Conclusion

CRARUM has the potential to strengthen the science of implementation research by addressing the complexities of practice settings, and engaging potential adopters to critically reflect on existing and proposed practices and strategies for sustaining change.     

Quantitative analysis of State 1–State 2 transitions in intact leaves using modulated fluorimetry — evidence for changes in the absorption cross-section of the two photosystems during state transitions

Using a combination of modulated and non-modulated light with synchronized detection it has been possible to monitor State 1–State 2 transitions in intact leaves as changes in the yield of modulated chlorophyll fluorescence. In the presence of excess far-red non-modulated light (713 nm) absorbed mainly by Photosystem I (PS I), the modulated fluorescence intensity was taken to represent F_o — the emission yield which occurs when the reaction centres of Photosystem II (PS II) are all open. On the other hand, superimposing saturating non-modulated wide-band, blue-green light resulted in a transitory maximum yield of modulated chlorophyll fluorescence, F_m, due to the total closure of the PS II reaction centres. In the absence of these additional lights the fluorescence level assumed a steady-state value, F_s, between F_o and F_m. All these parameters changed as the leaf slowly adapted to light of a given spectral composition. It was found that both F_o and F_m increased reversibly (by about 15–20%) during the transition from State 2 to State 1 such that the ratio of F_m to F_o remained constant, indicative of changes in absorption cross-section of PS II and PS I rather than alterations in ‘spillover’ which would cause preferential changes in F_m. It was also possible to estimate the fractions of light, β and α, channeled to PS II and PS I, respectively, from the values of F_o, F_m and F_s. In one approach, β was estimated in State 1, using the assumption that α + β = 1, and its variation during the subsequent state transition was assumed to follow proportional changes in F_o (or F_m). It was found that in State 2 there is a small loss (about 4%) of the total utilization of light in both photosystems. However, if such loss is neglected, assuming α + β is always unity, the calculated β was found to vary in the same direction and almost with the same magnitude as F_o (or F_m), indicating independently that a change in absorption cross-section in PS II (and PS I) had occurred. Consistent with these data were the light-saturation curves for the non-modulated far-red light-quenching effect in bringing the fluorescence from F_s to F_o in States 1 and 2. The ratio of the initial slopes of these curves indicates quantitatively both redistribution of light between PS I and PS II during the State 1–State 2 transitions and a partial loss of excitation energy in State 2.