Retinal-Carotenoid Interactions in a Sodium-Ion-Pumping Rhodopsin: Implications on Oligomerization and Thermal Stability.

Microbial rhodopsin (also called retinal protein)-carotenoid conjugates represent a unique class of light-harvesting (LH) complexes, but their specific interactions and LH properties are not completely elucidated as only few rhodopsins are known to bind carotenoids. Here, we report a natural sodium-ion (Na+)-pumping Nonlabens (Donghaeana) dokdonensis rhodopsin (DDR2) binding with a carotenoid salinixanthin (Sal) to form a thermally stable rhodopsin-carotenoid complex. Different spectroscopic studies were employed to monitor the retinal-carotenoid interaction as well as the thermal stability of the protein, while size-exclusion chromatography (SEC) and homology modeling are performed to understand the protein oligomerization process. In analogy with that of another Na+-pumping protein Krokinobacter eikastus rhodopsin 2 (KR2), we propose that DDR2 (studied concentration range: 2 × 10-6 to 4 × 10-5 M) remains mainly as a pentamer at room temperature and neutral pH, while heating above 55 °C partially converted it into a thermally less stable oligomeric form of the protein. This process is affected by both the pH and concentration. At high concentrations (4 × 10-5 to 2 × 10-4 M), the protein adopts a pentamer form reflected in the excitonic circular dichroism (CD) spectrum. In the presence of Sal, the thermal stability of DDR2 is increased significantly, and the pigment is stable even at 85 °C. The results presented could have implications in designing stable rhodopsin-carotenoid antenna complexes.

Bidirectional Photochemistry of Antarctic Microbial Rhodopsin: Emerging Trend of Ballistic Photoisomerization from the 13-cis Resting State.

The decades-long ultrafast examination of nearly a dozen microbial retinal proteins, ion pumps, and sensory photoreceptors has not identified structure-function indicators which predict photoisomerization dynamics, whether it will be sub-picosecond and ballistic or drawn out with complex curve-crossing kinetics. Herein, we report the emergence of such an indicator. Using pH control over retinal isomer ratios, photoinduced transient absorption is recorded in an inward proton pumping Antarctic microbial rhodopsin (AntR) for 13-cis and all-trans retinal resting states. The all-trans fluorescent state decays with 1 ps exponential kinetics. In contrast, in 13-cis it decays within ∼300 fs accompanied by continuous spectral evolution, indicating ballistic internal conversion. The coherent wave packet nature of 13-cis isomerization in AntR matches published results for bacteriorhodopsin (BR) and Anabaena sensory rhodopsin (ASR), which also accommodate both all-trans and 13-cis retinal resting states, marking the emergence of a first structure-photodynamics indicator which holds for all three tested pigments.

DEER experiments reveal fundamental differences between calmodulin complexes with IQ and MARCKS peptides in solution.

Calmodulin (CaM) is a calcium-binding protein that regulates the function of many proteins by indirectly conferring Ca2+ sensitivity, and it undergoes a large conformational change on partners' binding. We compared the solution binding mode of the target peptides MARCKS and IQ by double electron-electron resonance (DEER) distance measurements and paramagnetic NMR. We combined nitroxide and Gd(III) spin labels, including specific substitution of one of the Ca2+ ions in the CaM mutant N60D by a Gd(III) ion. The binding of MARCKS to holo-CaM resulted neither in a closed conformation nor in a unique relative orientation between the two CaM domains, in contrast with the crystal structure. Binding of IQ to holo-CaM did generate a closed conformation. Using elastic network modeling and 12 distance restraints obtained from multiple holo-CaM/IQ DEER data, we derived a model of the solution structure, which is in reasonable agreement with the crystal structure.

Kidney and blood pressure abnormalities 6 years after acute kidney injury in critically ill children: a prospective cohort study.

BACKGROUND: Acute kidney injury (AKI) in pediatric intensive care unit (PICU) children may be associated with long-term chronic kidney disease or hypertension. OBJECTIVES: To estimate (1) prevalence of kidney abnormalities (low estimated glomerular filtration rate (eGFR) or albuminuria) and blood pressure (BP) consistent with pre-hypertension or hypertension, 6 years after PICU admission; (2) if AKI is associated with these outcomes. METHODS: Longitudinal study of children admitted to two Canadian PICUs (January 2005-December 2011). Exposures (retrospective): AKI or stage 2/3 AKI (KDIGO creatinine-based definition) during PICU. Primary outcome (single visit 6 years after admission): presence of (a) low eGFR (<90 ml/min/1.73 m2) or albuminuria (albumin to creatinine ratio >30 mg/g) (termed "CKD signs") or (b) BP consistent with ≥pre-hypertension (≥90th percentile) or hypertension (≥95th percentile). RESULTS: Of 277 children, 25% had AKI. AKI and stage 2/3 AKI were associated with 2.2- and 6.6-fold higher adjusted odds, respectively, for the 6-year outcomes. Applying new hypertension guidelines attenuated associations; stage 2/3 AKI was associated with 4.5-fold higher adjusted odds for 6-year CKD signs or ≥elevated BP. CONCLUSIONS: Kidney and BP abnormalities are common 6 years after PICU admission and associated with AKI. Other risk factors must be elucidated to develop follow-up recommendations and reduce cardiovascular risk.

Cardiac Surgery-Associated Kidney Injury in Children and Renal Oximetry.

OBJECTIVES: Cardiac surgery-associated acute kidney injury is common in children and associates with negative outcomes. Novel interventions to reduce cardiac surgery-associated acute kidney injury require knowledge of its pathophysiology. States of altered perfusion, oxygen delivery, and energy consumption occur during cardiopulmonary bypass and could protect against or contribute to renal cellular injury and recovery. Near-infrared spectroscopy is noninvasive technology for monitoring regional blood flow and tissue oxygenation. This study evaluated the relationship between renal regional oxygen saturation and cardiac surgery-associated acute kidney injury, using near-infrared spectroscopy monitoring before, during, and after cardiopulmonary bypass in children. DESIGN: Prospective cohort study. SETTING: Single-center, tertiary care pediatric hospital (Stollery Children's Hospital, Edmonton, AB, Canada). PATIENTS: Children less than or equal to 10 kg undergoing congenital heart disease repair with cardiopulmonary bypass. Heart transplant, preoperative dialysis, sepsis, extracorporeal life support, congenital renal disease, and preoperative nephrotoxins were exclusions. INTERVENTIONS: Renal regional near-infrared spectroscopy monitoring before, during, and after cardiopulmonary bypass. MEASUREMENTS AND MAIN RESULTS: Outcome measure was cardiac surgery-associated acute kidney injury (defined according to Kidney Disease: Improving Global Outcomes criteria). Regional oxygen saturation was measured continuously using near-infrared spectroscopy (INVOS 5100C Cerebral/Somatic Oximeter; Medronic, Troy, MI) from time of anesthesia to time of transfer to intensive care. Cardiac surgery-associated acute kidney injury occurred in 65%. Lower baseline (precardiopulmonary bypass) regional oxygen saturation was associated with decreased risk of cardiac surgery-associated acute kidney injury (p = 0.01); children with baseline regional oxygen saturation in the highest tertile were 7.14 times more likely to get cardiac surgery- associated acute kidney injury (vs lowest tertile). Area under the curve for ability of baseline regional oxygen saturation to predict cardiac surgery-associated acute kidney injury was 0.73 (95% CI, 0.60-0.85). Children with lower baseline glomerular filtration rate had lower mean renal regional oxygen saturation. CONCLUSIONS: Findings demonstrate that preoperative oxygen supply/demand balance is an important predictor of cardiac surgery-associated acute kidney injury, suggesting lower preoperative (and intraoperative) renal blood flow may be protective. There is not yet a definite link between remote ischemic preconditioning and prevention of cardiac surgery-associated acute kidney injury; however, renal protective effects of sublethal ischemia should continue to be explored.

A high constitutive catalase activity confers resistance to methyl viologen-promoted oxidative stress in a mutant of the cyanobacterium Nostoc punctiforme ATCC 29133.

A spontaneous methyl viologen (MV)-resistant mutant of the nitrogen-fixing cyanobacterium Nostoc punctiforme ATCC 29133 was isolated and the major enzymatic antioxidants involved in combating MV-induced oxidative stress were evaluated. The mutant displayed a high constitutive catalase activity as a consequence of which, the intracellular level of reactive oxygen species in the mutant was lower than the wild type (N. punctiforme) in the presence of MV. The superoxide dismutase (SOD) activity that consisted of a SodA (manganese-SOD) and a SodB (iron-SOD) was not suppressed in the mutant following MV treatment. The mutant was, however, characterised by a lower peroxidase activity compared with its wild type, and its improved tolerance to externally added H2O2 could only be attributed to enhanced catalase activity. Furthermore, MV-induced toxic effects on the wild type such as (1) loss of photosynthetic performance assessed as maximal quantum yield of photosystem II, (2) nitrogenase inactivation, and (3) filament fragmentation and cell lysis were not observed in the mutant. These findings highlight the importance of catalase in preventing MV-promoted oxidative damage and cell death in the cyanobacterium N. punctiforme. Such oxidative stress resistant mutants of cyanobacteria are likely to be a better source of biofertilisers, as they can grow and fix nitrogen in an unhindered manner in agricultural fields that are often contaminated with the herbicide MV, also commonly known as paraquat.

Azide-functionalized lanthanide-based metal-organic frameworks showing selective CO2 gas adsorption and postsynthetic cavity expansion.

We report herein selective CO2 gas adsorption by two azide-functionalized lanthanide-based metal-organic frameworks (MOFs). This work also demonstrates that azide-functionalized MOFs can be used for postsynthetic cavity expansion, further corroborated by enhanced gas-sorption data.

The rates of charge separation and energy destructive charge recombination processes within an organic dyad in presence of metal-semiconductor core shell nanocomposites.

Steady state and time resolved spectroscopic measurements were made at the ambient temperature on an organic dyad, 1-(4-Chloro-phenyl)-3-(4-methoxy-naphthalen-1-yl)-propenone (MNCA), where the donor 1-methoxynaphthalene (1 MNT) is connected with the acceptor p-chloroacetophenone (PCA) by an unsaturated olefinic bond, in presence of Ag@TiO2 nanoparticles. Time resolved fluorescence and absorption measurements reveal that the rate parameters associated with charge separation, k(CS), within the dyad increases whereas charge recombination rate k(CR) reduces significantly when the surrounding medium is changed from only chloroform to mixture of chloroform and Ag@TiO2 (noble metal-semiconductor) nanocomposites. The observed results indicate that the dyad being combined with core-shell nanocomposites may form organic-inorganic nanocomposite system useful for developing light energy conversion devices. Use of metal-semiconductor nanoparticles may provide thus new ways to modulate charge recombination processes in light energy conversion devices. From comparison with the results obtained in our earlier investigations with only TiO2 nanoparticles, it is inferred that much improved version of light energy conversion device, where charge-separated species could be protected for longer period of time of the order of millisecond, could be designed by using metal-semiconductor core-shell nanocomposites rather than semiconductor nanoparticles only.

Is dye mixture more suitable rather than single dye to fabricate dye sensitized solar cell?

The steady state and time resolved spectroscopic studies reveal that two xanthene dyes Rhodamine 6G (R6G) and Rhodamine B (RB), used in the present investigations, form ground state hydrogen -bonded complexes with meso-tetrakis(4-carboxyphenyl) porphyrin (TCPP). However, it is apparent that upon photoexcitation the H-bonding complexes formed in the ground state decompose into the individual reacting components. This presumption was confirmed from the observation of the presence of only static quenching mode in the steady state fluorescence of the dyes in presence of porphyrin. The photoelectrochemical properties of the free dyes and the mixtures of each dye with porphyrin are investigated by measuring incident photon-to-current conversion efficiency (IPCE) using ZnO electrode and also with TiO2 electrode. It is seen that Rhodamine B-porphyrin mixture has attained maximum IPCE among the four samples studied at approximately 550 nm using ZnO electrode. Using TiO2 electrode, slight improvement in the value of IPCE was found for the same mixture. Therefore Rhodamine B-porphyrin mixture may act as a good sensitizer for converting solar energy to electrical energy.

Detailed spectroscopic investigations to reveal the nature of interaction of anionic porphyrin with calf thymus DNA.

The interaction between anionic form of meso-tetrakis(4-carboxyphenyl) porphyrin (TCPP) and calf thymus deoxyribonucleic acid (CT DNA) is investigated by measuring UV-vis absorption, steady-state fluorescence, steady-state fluorescence anisotropy, time-resolved fluorescence, resonance light scattering (RLS), FT-IR and circular dichroism (CD) spectra along with the help of atomic force microscopy (AFM) under Tris-Borate-EDTA (TBE) buffer solution of pH 8.3. The static mode of fluorescence quenching of porphyrin by calf thymus deoxyribonucleic acid indicates the formation of a ground-state complex. The formation of ground-state complex is a spontaneous molecular interaction procedure in which outside groove binding through hydrogen bond or van der Waals force plays a major role. For biomedical application this investigation is very important as here TCPP, i.e. the anionic porphyrin does not bring any changes in the original structure of the CT DNA to selectively cleaving the nucleic acid to destroy the cancer or tumor cells whereas cationic porphyrin makes change in the protein structure significantly during the same process.

Development of a nanocomposite system by combining an organic dyad 1-(4-chloro-phenyl)-3-(4-methoxy-naphthalen-1-yl)-Propenone with semiconductor TiO2 nanoparticles.

Steady state and time resolved spectroscopic measurements on an organic dyad, 1-(4-chlorophenyl)-3-(4-methoxy-naphthalen-1-yl)-propenone, where the donor 1-methoxynaphthalene is connected with the acceptor p-chloroacetophenone by an unsaturated olefinic bond, in presence of TiO2 nanoparticles were made at the ambient temperature. Time resolved fluorescence measurements reveal that the rate parameters associated with charge separation, kCS, within the dyad increases whereas charge recombination rate reduces when the surrounding medium is changed from chloroform to TiO2 nanoparticles. The observed results indicate that the dyad being combined with TiO2 nanoparticles may form organic-inorganic nanocomposite systems useful for developing light energy conversion devices.

Nonradiative inter- and intramolecular energy transfer from the aromatic donor anisole to a synthesized photoswitchable acceptor system.

We report steady state and time resolved fluorescence measurements on acetonitrile (ACN) solutions of the model compounds, energy donor anisole (A) and a photoswitchable acceptor N,N'-1,2-phenylene di-p-tosylamide (B) and the multichromophore (M) where A and B are connected by a spacer containing both rigid triple (acetylenic) and flexible methylene bonds. Both steady state and time correlated single photon counting measurements demonstrate that though intermolecular energy transfer, of Forster type, between the donor and acceptor moieties occurs with rate 10(8)s(-1) but when these two reacting components are linked by a spacer (multichromophore, M) the observed transfer rate ( approximately 10(11)s(-1)) enhances. This seemingly indicates that the imposition of the spacer by inserting a triple bond may facilitate in the propagation of electronic excitation energy through bond. The time resolved fluorescence measurements along with the theoretical predictions using Configuration interaction singles (CIS) method by using 6-31G (d,p) basis set, implemented in the Gaussian package indicate the formations of the two excited conformers of B. The experimental findings made from the steady state and time resolved fluorescence measurements demonstrate that, though two different isomeric species of the acceptor B are formed in the excited singlet states, the prevailing singlet-singlet nonradiative energy transfer route was found from the donor A to the relatively longer-lived isomeric species of B.

N-hetero atomic effect on the photophysics of 2,2'-dipyridylketone.

Emission characteristics of 2,2'-dipyridylketone (DPK) have been studied in the rigid glass matrices of ethanol (EtOH) and methyl cyclohexane (MCH) at 77 K. As in the case of the aromatic analogue benzophenone (BOP), DPK is found to exhibit only phosphorescence emission. Although both BOP and DPK have the lowest triplet states chiefly of n pi* nature, the presence of other triplet states in the neighbourhood of the lowest triplet one is found to play some significant roles in determining the photophysical properties of DPK, specially in the non-polar solvent (MCH). All the photophysical properties including the polarization characteristics have been explored in both the molecules. Experimental observations are corroborated with necessary quantum chemical calculations.

Photophysical processes involved within the anisole-thioindoxyl dyad system.

The photophysical properties and the nature of the photoinduced electron transfer (PET) reactions within a synthesized anisole (A)-thioindoxyl (T) dyad system (24MBTO) have been studied by electrochemical, steady-state, and time-resolved spectroscopic techniques. Computations on the dyad were performed both in gas phase as well as solvent environment by TD-DFT method with B3LYP density function. The geometry optimization calculation of 24MBTO was done by 6-311G(d,p) basis function set implemented in the Gaussian package. The theoretical values of singlet vertical excitation energies were found to correlate well with the experimentally observed ones. The electrochemical measurements indicate the possibility of occurrence of PET reactions within 24MBTO between the linked redox centers A and T. Both steady-state and time-resolved spectroscopic measurements on the novel synthesized 24MBTO dyad demonstrate the formations of the two types of isomeric species: Z- and E- forms, resulted from the charge separation reactions. From the detailed studies it reveals that the present thioaurone may behave as a versatile photoswitchable system. It has been hinted that the loss process (charge recombination) within 24MBTO could possibly be prevented by incorporating it within the hydrophobic cavity of beta-cyclodextrin (betaCD).