Nasal nanovaccines.

Nasal administration of vaccines is convenient for the potential stimulation of mucosal and systemic immune protection. Moreover the easy accessibility of the intranasal route renders it optimal for pandemic vaccination. Nanoparticles have been identified as ideal delivery systems and adjuvants for vaccine application. Heterogeneous protocols have been used for animal studies. This complicates the understanding of the formulation influence on the immune response and the comparison of the different nanoparticles approaches developed. Moreover anatomical and immunological differences between rodents and humans provide an additional hurdle in the rational development of nasal nanovaccines. This review will give a comprehensive expertise of the state of the art in nasal nanovaccines in animals and humans focusing on the nanomaterial used.

Mechanisms allowing protein delivery in nasal mucosa using NPL nanoparticles.

The intranasal administration of proteins using nanoparticles is a promising approach for several applications, especially for mucosal vaccines. Delivery of protein within the epithelial barrier is a key point to elicit an immune response and nano-carrier has to show no toxicity. The aim of this work was to elucidate the interactions of cationic porous nanoparticles loaded with protein delivery for antigen delivery in the nose. We investigated the loading, the cellular delivery and the epithelial transcytosis of proteins associated to these nanoparticles containing an anionic lipid in their core (NPL). NPL were highly endocytosed by airway epithelial cells and significantly improved the protein delivery into the cell. In vitro transcytosis studies showed that NPL did not modify the in vitro epithelial permeability suggesting no toxicity of these carriers. Moreover protein and NPL did not translocate the epithelial barrier. In vivo studies demonstrated that NPL prolonged the nasal residence time of the protein and no NPL were found beyond the epithelial barrier in vivo, precluding a negative side effect. All together these results establish the NPL as a bio-eliminable and optimal vaccine carrier.

Development of innovative paclitaxel-loaded small PLGA nanoparticles: study of their antiproliferative activity and their molecular interactions on prostatic cancer cells.

Taxanes, including paclitaxel, are anti-cancer drugs approved for the treatment of prostate cancer but which have limited clinical application due to their hydrophobicity, their low therapeutic index and the emergence of chemoresistance. These side effects may be avoided through the use of new drug delivery systems such as nanoparticles, and paclitaxel-loaded PLGA nanoparticles up to 200 nm in size have shown encouraging results. As it is known that size affects the tissular penetration and distribution of tumors via the enhanced permeability and retention effect, so nanoparticles smaller than 100 nm are potentially interesting vehicles for improving paclitaxel delivery and efficacy. In this work, new paclitaxel-loaded small PLGA nanoparticles, between 49 nm and 95 nm in size and with positive or negative surface charges, were prepared without detergent. They were stable in the presence of serum, and HPLC showed that high paclitaxel loading and stability were achieved. Intracellular uptake of these nanoparticles was studied in PC3 cells by flow cytometry. Confocal studies confirmed a high tubulin destructuration at very low dose with these nanoparticles. This study suggests that both positively and negatively charged paclitaxel-loaded small PLGA nanoparticles deliver this drug into PC3 cells, and that this nanoparticle mode of delivery highly improves paclitaxel efficiency by up to two log-increase. These results also highlight the importance of small nanoparticles for drug delivery in cancer applications and are extremely promising for in vivo studies.

Binding of biogenic and synthetic polyamines to ß-lactoglobulin.

The bindings of biogenic polyamines spermine (spm), spermidine (spmd) and synthetic polyamines 3,7,11,15-tetrazaheptadecane·4HCl (BE-333) and 3,7,11,15,19-pentazahenicosane·5HCl (BE-3333) with ß-lactoglobulin (ß-LG) were determined in aqueous solution. FTIR, UV-vis, CD and fluorescence spectroscopic methods as well as molecular modeling were used to determine the polyamine binding sites and the effect of polyamine complexation on protein stability and secondary structure. Structural analysis showed that polyamines bind ß-LG via both hydrophilic and hydrophobic contacts. Stronger polyamine-protein complexes formed with synthetic polyamines than biogenic polyamines, with overall binding constants of K(spm-ß-LG)=3.2(±0.6)×10(4) M(-1), K(spmd-ß-LG)=1.8(±0.5)×10(4) M(-1), K(BE-333-ß-LG)=5.8(±0.3)×10(4) M(-1) and K(BE-3333-ß-LG)=6.2(±0.05)×10(4) M(-1). Molecular modeling showed the participation of several amino acids in the polyamine complexes with the following order of polyamine-protein binding affinity: BE-3333>BE-333>spermine>spermidine, which correlates with their positively charged amino group content. Alteration of protein conformation was observed with a reduction of ß-sheet from 57% (free protein) to 55-51%, and a major increase of turn structure from 13% (free protein) to ∼21% in the polyamine-ß-LG complexes, indicating a partial protein unfolding.

Polyamines interaction with thylakoid proteins during stress.

The involvement of polyamines in plant responses to abiotic stresses is well investigated, while there has been few reports on the specific mode of action of polyamines on the photosynthetic apparatus. The objective of this review is thus to examine the mode of interaction of polyamines with proteins of photosystem II core and LHCII, including methylamine (monoamine) as a simplified model to better understand the mode of action of polyamines. Spectroscopic methods used to determine the binding mode of amines with PSII proteins showed that amines such as spermine, putrescine and methylamine interact with protein (H-bonding) through polypeptide C=O, C-N and N-H groups with major perturbations of protein secondary structure as the concentration of amines was raised. High concentration of amines added to PSII-enriched submembrane fractions causes a significant loss of PSII activity. However, at lower concentration, polyamines, especially spermine, improve the photosynthetic functions under stress. We concluded from this review that besides the conjugation of polyamines with LHC polypeptides, polyamines are likely to interact with extrinsic proteins and the hydrophilic part of intrinsic proteins of PSII by electrostatic interaction. This could stabilize the conformation of proteins under various stresses. However, at high concentration of polyamines a strong inhibition of PSII activity is observed.

Methylamine interaction with proteins of photosystem II: a comparison with biogenic polyamines.

Biogenic polyamines are essential for cell growth and differentiation. The interaction of polyamines with protein of photosystem II (PSII) are well investigated, while there has been no report on the effect of monoamines complexation on photosynthetic oxygen evolution. This study was designed to investigate the interaction of methylamine with proteins of PSII, using PSII-enriched submembrane fractions with various concentrations of methylamine. Fourier transformed infrared (FTIR) and fluorescence spectroscopic methods were used in order to determine the methylamine binding mode, the protein conformational changes, and the effect of amine interaction on photosynthetic oxygen evolution. Spectroscopic evidence showed that methylamine interacts with protein (H-bonding) through polypeptide C=O, C-N and NH groups with major perturbations of protein secondary structure. Major reduction of alpha-helix from 50% (free PSII) to 35% with an increase of beta-sheet from 10% (free PSII) to 16% was observed in methylamine-PSII complexes. At very low methylamine concentration, no inhibition of oxygen-evolution occurred, while at higher amine content (12 mM), 100% inhibition was observed. Chlorophyll (Chl) fluorescence measurements indicated the inhibition mainly affects the oxygen evolving complex (OEC) of PSII. Comparisons of the effects of methylamine with biogenic polyamine spermine, spermidine and putrescine showed a similar mode of binding with protein (H-bonding) through polypeptide C=O, C-N and NH groups. However, major alterations of the protein secondary structure are induced by monoamine and not by polyamines.

Plant tissue-and photosynthesis-based biosensors.

Biosensors are promising biotools, alternative or complementary to conventional analysis techniques, for fast, simple, cheap and reliable screening. This article reviews the biosensors that use plant components as biorecognition elements. In the first section, plant tissue-based biosensors are summarised and classified according to the enzyme used. Afterwards, photosynthesis-based biosensors, including the types of photosynthetic materials and immobilisation methods, are described.

Polyamine analogues bind human serum albumin.

Polyamine analogues show antitumor activity in experimental models, and their ability to alter activity of cytotoxic chemotherapeutic agents in breast cancer is well documented. Association of polyamines with nucleic acids and protein is included in their mechanism of action. The aim of this study was to examine the interaction of human serum albumin (HSA) with several polyamine analogues, such as 1,11-diamino-4,8-diazaundecane (333), 3,7,11,15-tetrazaheptadecane.4HCl (BE-333), and 3,7,11,15,19-pentazahenicosane.5HCl (BE-3333), in aqueous solution at physiological conditions using a constant protein concentration and various polyamine contents (microM to mM). FTIR, UV-visible, and CD spectroscopic methods were used to determine the polyamine binding mode and the effects of polyamine complexation on protein stability and secondary structure. Structural analysis showed that polyamines bind nonspecifically (H-bonding) via polypeptide polar groups with binding constants of K333 = 9.30 x 10(3) M(-1), KBE-333 = 5.63 x 10(2) M(-1), and KBE-3333 = 3.66 x 10(2) M(-1). The protein secondary structure showed major alterations with a reduction of alpha-helix from 55% (free protein) to 43-50% and an increase of beta-sheet from 17% (free protein) to 29-36% in the 333, BE-333, and BE-3333 complexes, indicating partial protein unfolding upon polyamine interaction. HSA structure was less perturbed by polyamine analogues compared to those of the biogenic polyamines.

Resveratrol binding to human serum albumin.

Resveratrol (Res), a polyphenolic compound found largely in the skin of red grape and wine, exhibits a wide range of pharmaceutical properties and plays a role in prevention of human cardiovascular diseases [Pendurthi et al., Arterioscler. Thromb. Vasc. Biol. 19, 419-426 (1999)]. It shows a strong affinity towards protein binding and used as inhibitor for cyclooxygenase and ribonuclease reductase. The aim of this study was to examine the interaction of resveratrol with human serum albumin (HSA) in aqueous solution at physiological conditions, using a constant protein concentration (0.3 mM) and various pigment contents (microM to mM). FTIR, UV-Visible, CD, and fluorescence spectroscopic methods were used to determine the resveratrol binding mode, the binding constant and the effects of pigment complexation on protein secondary structure. Structural analysis showed that resveratrol bind non-specifically (H-bonding) via polypeptide polar groups with overall binding constant of K(Res) = 2.56 x 10(5) M(-1). The protein secondary structure, analysed by CD spectroscopy, showed no major alterations at low resveratrol concentrations (0.125 mM), whereas at high pigment content (1 mM), major increase of alpha-helix from 57% (free HSA) to 62% and a decrease of beta-sheet from 10% (free HSA) to 7% occurred in the resveratrol-HSA complexes. The results indicate a partial stabilization of protein secondary structure at high resveratrol content.

The effects of drug complexation on the stability and conformation of human serum albumin: protein unfolding.

We report different analytical methods used to study the effects of 3\'-azido-3\'-deoxythymidine, aspirin, taxol, cisplatin, atrazine, 2,4-dichlorophenoxyacetic, biogenic polyamines, chlorophyll, chlorophyllin, poly(ethylene glycol), vanadyl cation, vanadate anion, cobalt-hexamine cation, and As2O3, on the stability and secondary structure of human serum albumin (HSA) in aqueous solution, using capillary electrophoresis, Fourier transform infrared, ultraviolet visible, and circular dichroism (CD) spectroscopic methods. The concentrations of HSA used were 4% to 2% or 0.6 to 0.3 mM, while different ligand concentrations were 1 microM to 1 mM. Structural data showed drugs are mostly located along the polypeptide chains with both specific and nonspecific interactions. The stability of drug-protein complexes were in the order K(VO(2+)) 1.2 x 10(8) M(-1) > K(AZT) 1.9 x 10(6) M(-)1 > K(PEG) 4.1 x 10(5) M(-1) > K(atrazine) 3.5 x 10(4) M(-1) > K(chlorophyll) 2.9 x 10(4) M(-1) > K2,4-D 2.5 x 10(4) M-1 > K(spermine) 1.7 x 10(4) M(-1) > K(taxol) 1.43 x 10(4) M(-1) > K(Co(3+)) > 1.1 x 10(4) M(-1) > K(aspirin) 1.04 x 10(4)i(-1) > K(chlorophyllin) 7.0 x 10(3) M(-1) > K(VO(3)(-)) 6.0 x 103 M(-1) > K(spermidine) 5.4 x 10(3) M(-1) > K(putrescine) 3.9 x 10(3) M(-1) > K(As(2)O(3)) 2.2 x 10(3) M(-1)> K(cisplatin) 1.2 x 10(2) M(-1). The protein conformation was altered (infrared and CD results) with major reduction of alpha-helix from 60 to 55% (free HSA) to 49 to 40% and increase of beta-structure from 22 to 15% (free HSA) to 33 to 23% in the drug-protein complexes. The alterations of protein secondary structure are attributed to a partial unfolding of HSA on drug complexation.

Effects of organic and inorganic polyamine cations on the structure of human serum albumin.

The presence of several high affinity binding sites on human serum albumin (HSA) makes it a possible target for many organic and inorganic molecules. Organic polyamines are widely distributed in living cells and their biological roles have been associated with their physical and chemical interactions with proteins, nucleic acids, and lipids. This study is designed to examine the effects of spermine, spermidine, putrescine, and cobalt [Co(III)]-hexamine cations on the solution structure of HSA using Fourier transform IR, UV-visible, and circular dichroism (CD) spectroscopic methods. The spectroscopic results show that polyamine cations are located along the polypeptide chains with no specific interaction. The order of perturbations is associated with the number of positive charges of the polyamine cation: spermine > Co(III)-hexamine > spermidine > putrescine. The overall binding constants are 1.7 x 10(4), 1.1 x 10(4), 5.4 x 10(3), and 3.9 x 10(3)M(-1), respectively. The protein conformation is altered (IR and CD data) with reductions of alpha helices from 60 to 55% for free HSA to 50-40% and with increases of beta structures from 22 to 15% for free HSA to 33-23% in the presence of polyamine cations.

Secondary structural analysis of the Na(+),K(+)-ATPase and its Na(+) (E(1)) and K(+) (E(2)) complexes by FTIR spectroscopy.

The Na(+),K(+)-ATPase is an integral membrane protein which transports sodium and potassium cations against an electrochemical gradient. The transport of Na(+) and K(+) ions is presumably connected to an oscillation of the enzyme between the two conformational states, the E(1) (Na(+)) and the E(2) (K(+)) conformations. The E(1) and E(2) states have different affinities for ligand interaction. However, the determination of the secondary structure of this enzyme in its sodium and potassium forms has been the subject of much controversy. This study was designed to provide a quantitative analysis of the secondary structure of the Na(+),K(+)-ATPase in its sodium (E(1)) and potassium (E(2)) states in both H(2)O and D(2)O solutions at physiological pH, using Fourier transform infrared (FTIR) with its self-deconvolution and second derivative resolution enhancement methods, as well as curve-fitting procedures. Spectroscopic analysis showed that the secondary structure of the sodium salt of the Na(+),K(+)-ATPase in H(2)O solution contains alpha-helix 19.8+/-1%, beta-sheet 25.6+/-1%, turn 9.1+/-1%, and beta-anti 7.5+/-1%, whereas in D(2)O solution, the enzyme shows alpha-helix 16.8+/-1%, beta-sheet 24.5+/-1.5%, turn 10.9+/-1%, beta-anti 9.8+/-1%, and random coil 38.0+/-2%. Similarly, the potassium salt in H(2)O solution contains alpha-helix 16.6+/-1%, beta-sheet 26.4+/-1.5%, turn 8.9+/-1%, and beta-anti 8.1+/-1%, while in D(2)O solution it shows alpha-helix 16.2+/-1%, beta-sheet 24.5+/-1.5%, turn 10.3+/-1%, beta-anti 9.0+/-1%, and random coil 40+/-2%. Thus the main differences for the sodium and potassium forms of the Na(+),K(+)-ATPase are alpha-helix 3.2% in H(2)O and 0.6% in D(2)O, beta-sheet (pleated and anti) 1.5% in H(2)O and random structure 2% (D(2)O), while for other minor components (turn structure), the differences are less than 1%.

Changes in the structure of chlorophyll-protein complexes and excitation energy transfer during photoinhibitory treatment of isolated photosystem I submembrane particles.

The activity of light-induced oxygen consumption, absorption spectra, low temperature (77 K) chlorophyll fluorescence emission and excitation spectra were studied in suspensions of photosystem (PS) I submembrane particles illuminated by 2000 microE m(-2) s(-1) strong white light (WL) at 4 degrees C. A significant stimulation of oxygen uptake was observed during the first 1-4 h of photoinhibitory treatment, which rapidly decreased during further light exposure. Chlorophyll (Chl) content gradually declined during the exposure of isolated PSI particles to strong light. In addition to the Chl photobleaching, pronounced changes were found in Chl absorption and fluorescence spectra. The position of the major peak in the red part of the absorption spectrum shifted from 680 nm towards shorter wavelengths in the course of strong light exposure. A 6-nm blue shift of that peak was observed after 5-h illumination. Even more pronounced changes were found in the characteristics of Chl fluorescence. The magnitude of the dominating long-wavelength emission band at 736 nm located in untreated particles was five times reduced after 2-h exposure, whereas the loss in absolute Chl contents did not exceed 10% of its initial value. The major peak in low-temperature Chl fluorescence emission spectra shifted from 736 to 721 nm after 6-h WL treatment. Individual Chl-protein complexes differed in the response of their absorption spectra to strong WL. Unlike light-harvesting complexes (LHC), LHCI-680 and LHC-730, which did not exhibit changes in the major peak position, its maximum was shifted from 678 to 671 nm in CPIa complex after PSI submembrane particles were irradiated with strong light for 6 h. The results demonstrated that excitation energy transfer represents the stage of photosynthetic utilization of absorbed quanta which is most sensitive to strong light in isolated PSI particles.

Nonphotosynthetic reduction of the intersystem electron transport chain of chloroplasts following heat stress. The pool size of stromal reductants.

The properties of a negative transient signal (negative peak) observed during the first seconds of the induction of the photoacoustic (PA) signal in dark-adapted barley leaves treated with methyl viologen (MV) and diuron and then exposed to high temperatures have been examined. Under those conditions no electron donation from photosystem II (PSII) occurred, and electron flow through PSI could be supported only by soluble reductants located in the chloroplast stroma. The negative peak was observed only if the PA signal had been monitored at low, and not high, frequencies. The peak obviously originated from the oxygen consumption by PSI. The size of the peak increased as the temperature of preheating was raised from 39 to 45 degrees C. The size of the peak decreased exponentially with a half-time of 3.7 s during illumination under low light. This decrease was found to be much faster under strong light. The recovery of the peak during dark acclimation required several minutes. It is concluded that the negative peak reflects the oxygen consumption supported by stromal reductants, their pool being rapidly exhausted under light in the presence of MV. The maximal size of the pool was calculated as 140 eq: P700 in dark-adapated leaves.

A simple mediatorless amperometric method using the cyanobacterium Synechococcus leopoliensis for the detection of phytotoxic pollutants.

The unicellular cyanobacterium Synechoccocus leopoliensis is used in a micro-electrochemical cell to generate photocurrents. The photocurrent is dependent on photosynthetic electron transport and is mediated by hydrogen peroxide formation following the reduction of oxygen on the acceptor side of photosystem I. This is the first known application of cyanobacteria in an electrochemical device where no artificial electroactive mediator is needed. The potential for the development of this micro-electrochemical cell for the detection of phytotoxic pollutants, such as herbicides and toxic metal cations, using the photosynthetic system of the cyanobacteria without interference from added electron acceptor is discussed.

Interactions of atrazine and 2,4-D with human serum albumin studied by gel and capillary electrophoresis, and FTIR spectroscopy.

The herbicides 6-chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4-diamine (atrazine) and 2,4-dichlorophenoxyacetic acid (2,4-D) are widely used in agricultural practice to fight dicotyledon weeds mainly in maize, cereals, and lucerne. As a result, these compounds are found not only in the plants, soil, and water, but also in the cultivated ground in the following years as well as in agricultural products such as fruits, milk, butter, and sugar beet. The toxicological effects of herbicides occur in vivo, when transported to the target organ through the bloodstream. It has been suggested that human serum albumin (HSA) serves as a carrier protein to transport 2,4-D to molecular targets. This study was designed to examine the interaction of atrazine and 2,4-D with HSA in aqueous solution at physiological pH with herbicide concentrations of 0.0001-1 mM, and final protein concentration of 1% w/v. Gel and capillary electrophoresis, UV-visible and Fourier transform infrared spectroscopic methods were used to determine the drug binding mode, the drug binding constant, and the protein secondary structure in aqueous solution. Structural analysis showed that different types of herbicide-HSA complexes are formed with stoichiometric ratios (drug/protein) of 3:1 and 11:1 for atrazine and 4.5:1 and 10:1 for 2,4-D complexes. Atrazine showed a weak binding affinity (K=3.50 x 10(4) M(-1)), whereas two bindings (K(1)=2.50 x 10(4) M(-1) and K(2)=8.0 x 10(3) M(-1)) were observed for 2,4-D complexes. The herbicide binding results in major protein secondary structural changes from that of the alpha-helix 55% to 45--39% and beta-sheet 22% to 24--32%, beta-anti 12% to 10--22% and turn 11% to 12--15%, in the drug-HSA complexes. The observed spectral changes indicate a partial unfolding of the protein structure, in the presence of herbicides in aqueous solution.

Heterogeneity of Photosystem I reaction centers in barley leaves as related to the donation from stromal reductants.

The light-response curves of P700 oxidation and time-resolved kinetics of P700(+) dark re-reduction were studied in barley leaves using absorbance changes at 820 nm. Leaves were exposed to 45 degrees C and treated with either diuron or diuron plus methyl viologen (MV) to prevent linear electron flow from PS II to PS I and ferredoxin-dependent cyclic electron flow around PS I. Under those conditions, P700(+) could accept electrons solely from soluble stromal reductants. P700 was oxidized under weak far-red light in leaves treated with diuron plus MV, while identical illumination was nearly ineffective in diuron-treated leaves in the absence of MV. When heat-exposed leaves were briefly illuminated with strong far-red light, which completely oxidized P700, the kinetics of P700(+) dark reduction was fitted by a single exponential term with half-time of about 40 ms. However, two first-order kinetic components of electron flow to P700(+) (fast and slow) were found after prolonged leaf irradiation. The light-induced modulation of the kinetics of P700(+) dark reduction was reversed following dark adaptation. The fast component (half time of 80-90 ms) was 1.5 larger than the slow one (half time of about 1 s). No kinetic competition occurred between two pathways of electron donation to P700(+) from stromal reductants. This suggests the presence of two different populations of PS I.

Degradation of the photosystem I complex during photoinhibition.

Exposure of isolated photosystem I (PSI) complexes to illumination (2300 microE m(-2) s(-1)) for various periods of time resulted in striking changes in their absorption spectra. A 6 nm blueshift of the absorption maximum in the red was detected after 100 min illumination. The fourth derivative of the absorption spectra verifies that the main change of the red peak was attributed to the 682 nm absorption band. Further, it was also shown that a shoulder in the absorption spectra located around 470 nm decreased after the first 5 min of illumination and almost disappeared after 40 min illumination, suggesting that chlorophyll b bound to light-harvesting complex I (LHCI) is also sensitive to excess light. A maximum inhibitory effect on the oxygen uptake rates and a strong stimulation were observed when the PSI complexes were exposed to illumination for about 20 and 40 min, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that LHCI-680 started to degrade during the first 5 min of illumination and almost completely disappeared after 40 min of illumination. These observations demonstrated that LHCI was more sensitive to illumination than the PsaA/B subunits which also presented some degradation signs after 40 min illumination. In addition, insoluble-cohesive-denatured proteins also appeared between the stacking and resolving gel after prolonged illumination (100 min). A photoprotective function of LHCI for the PSI reaction center is proposed.

Nonphotosynthetic reduction of the intersystem electron transport chain of chloroplasts following heat stress. Steady-state rate.

The consequence of elevated temperatures in the range of 39-51 degrees C on the steady-state rate of light-induced electron transport through photosystem I (PSI) supported by stromal reductants was studied in intact barley leaves using photoacoustic and chlorophyll fluorescence techniques. Measurable electron flow through PSI in diuron-treated leaves occurred only after exposure to temperatures above 37 degrees C. The steady-state rate of the above diuron-insensitive electron flow with methyl viologen as electron acceptor was estimated to be 3.7 mu eq m-2 s-1 or 0.018 mu eq mumol chlorophyll-1 s-1 in leaves exposed for 5 min to 45 degrees C.

Redox characteristics of manganese and cobalt complexes obtained from pyridine N-oxide.

Manganese and cobalt complexes, using pyridine N-oxide as ligand, have been synthesized, and their cyclic and square-wave voltammetric measurements have been carried out. The results reveal that the complexes exhibit different voltammetric pattern, which suggests that the redox processes are most probably metal-centered. In both complexes, extra redox activity is observed once the potential exceeds certain value of the voltage. The observation of an oxidation wave in manganese complex at + 0.75 V vs. Ag/AgCl or + 0.95 V vs. NHE strongly suggests that this complex can bring about oxidation of water and can, thus, serve as a synthetic analogue of water oxidizing complex (WOC) of PS II.