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B Kok

Publications and source records attributed to B Kok.

At least 19 recordsLinked to original sources
PubMed1995

Nitrite and nitrate determinations in plasma: a critical evaluation.

Plasma nitrite and nitrate determinations are increasingly being used in clinical chemistry as markers for the activity of nitric oxide synthase and the production of nitric oxide radicals. However, a systematic evaluation of the determination of nitrite and nitrate in plasma has not been performed. In this study the recovery and stability of nitrite and nitrate in whole blood and in plasma, the relation between nitrite and nitrate concentrations in plasma, and possible sources of artifacts were investigated. The main conclusions are: (a) Recovery of nitrite and nitrate from plasma is near-quantitative (87%) and reproducible; (b) nitrite and nitrate are stable in (frozen) plasma for at least 1 year; (c) nitrite in whole blood is very rapidly (> 95% in 1 h) oxidized to nitrate, and therefore plasma nitrite determination alone is meaningless; (d) the ranges of nitrite and nitrate concentrations in plasma samples of 26 healthy persons are 1.3-13 mumol/L (mean 4.2 mumol/L) and 4.0-45.3 mumol/L (mean 19.7 mumol/L), respectively; (e) plasma nitrite and nitrate concentrations were not correlated (nitrite as % of total nitrite + nitrate varied from 3.9% to 88% in plasma samples); and (f) plasma samples should be deproteinized, and background controls for each sample should be included in the assay, to avoid measuring artifactually high nitrite and nitrate concentrations in plasma.

Drug Stability↗
PubMed1979

Redox titration of electron acceptor Q and the plastoquinone pool in photosystem II.

The primary photochemical quencher Q and the secondary electron acceptor pool in Photosystem II have been titrated. We used particles of Scenedesmus mutant No. 8 that lack System I and allowed the system to equilibrate with external redox mediators in darkness prior to measurement of the fluorescence rise curve. The titration of Q, as indicated by the dark level of Fi, occurs in two discrete steps. The high-potential component (Qh) has a midpoint potential of +68 mV (pH 7.2) and accounts for approximately 67% of Q. The pH sensitivity of the midpoint potential is -60 mV, indicating the involvement of 1 H+/e. The low-potential component (Q1) accounts for the remaining 33% of Q and shows a midpoint potential near--300 mV (pH 7.2). The plastoquinone pool, assayed as the half-time of the fluorescence rise curve, titrates as a single component with a midpoint potential 30--40 mV more oxidizing than that of Qh, i.e., at 106 mV (pH 7.2). The Em shows a pH sensitivity of -60 mV/pH unit, indicating the involvement of 1 H+/e. The observation that all 12--14 electron equivalents in the pool titrate as a single component indicates that the heterogeneity otherwise observed in the secondary acceptor system is a kinetic rather than a thermodynamic property. Illumination causes peculiar, and as yet unclarified, changes of both Q and the secondary pool under anaerobic conditions that are reversed by oxygen.

Chlorophyta↗
PubMed1978

Evidence that the intermediate electron acceptor, A2, in photosystem I is a bound iron-sulfur protein.

Absorption changes accompanying the formation of light-induced P-700+ were investigated in a highly enriched Photosystem I preparation where an intermediate electron acceptor preceding P-430 could be detected. In an enriched Photosystem I particle, light-induced reversible absorption changes observed at 700 nm in the presence of dithionite resembled those previously seen at 703 nm and 820 nm [9], thus indicating the presence of a backreaction between P700+ and A-2. After this same Photosystem I particle was treated to denature the bound iron-sulfur centers, the photochemical changes that could be attributed to P-700 in equilibrium A2 were completely lost. These results provide evidence that the intermediate electron acceptor, A2, is a bound iron-sulfur protein. Additional studies in the 400--500 nm region with Photosystem I particles prepared by sonication indicate that the spectrum of A2 is different from that of P-430.

Cytochromes↗
PubMed1978

Photosynthetic oxygen evolution from hydrogen peroxide.

A study was made of the interactions of flash-illuminated chloroplasts with hydrogen peroxide. We conclude: 1. The oxygen precursor system can be reduced beyond the S0 state to an S-1 state, which can be oxidized to S0 by a single flash. 2. In the dark, a two-electron donation by H2O2 takes place which reduces S2 to S0 and S1 to S-1. 3. At the same time, two-electron oxidations by H2O2 re-form, S2 from S0 and S1 from S-1. 4. The catalase-like activity due to this cyclic oxidation and reduction of the S enzyme is higher with the S2 in equilibrium S0 couple than with the S1 in equilibrium S-1 couple. Another process, however, is responsible for most of the O2 evolution from H2O2 in the light. Our evidence indicates that this process: (1) is independent of the S states and insensitive to Tris washing, (2) turns over rapidly in high concentrations of peroxide, (3) yields 1 O2 per electron passing through system II; (4) dismutates two H2O2 molecules, so that there is no net consumption of 'holes'.

Catalase↗
PubMed1976

Stabilization by glutaraldehyde of high-rate electron transport in isolated chloroplasts.

Treatment of isolated chloroplasts with glutaraldehyde affects their ability to photoreduce artificial electron acceptors. The remaining rate of O2 evolution approaches zero with methyl viologen, is low with ferricyanide, but nearly normal with lipophilic Photosystem II acceptors, like oxidized p-phenylenediamine and oxidized diaminodurene. Since Photosystem I donor reactions are also affected, a specific site of inhibition of electron transport to Photosystem I is indicated. At the same time, glutaraldehyde prolongs the longevity of the chloroplasts stored in dark. In control samples the half-life of Photosystem II activity varied between 5 days at 4degreesC and 1 day at 25degreesC. Glutaraldehyde treatment increased these half times approx. 3-fold. The glutaraldehyde doses required to induce inhibition and stabilization were very similar.

Aldehydes↗
PubMed1976

Determination of H+/e- ratios in chloroplasts with flashing light.

Using a rapid pH electrode, measurements were made of the flash-induced proton transport in isolated spinach chloroplasts. To calibrate the system, we assumed that in the presence of ferricyanide and in steady-state flashing light, each flash liberates from water one proton per reaction chain. We concluded that with both ferricyanide and methylviologen as acceptors two protons per electron are translocated by the electron transport chain connecting Photosystem II and I. With methyl viologen but not with ferricyanide as an acceptor, two additional protons per electron are taken up due to Photosystem I activity. One of these latter protons is translocated to the inside of the thylakoid while the other is taken up in H2O2 formation. Assuming that the proton released during water splitting remains inside the thylakoid, we compute H+/e- ratios of 3 and 4 for ferricyanide and methylviologen, respectively. In continuous light of low intensity, we obtained the same H+/e- ratios. However, with higher intensities where electron transport becomes rate limited by the internal pH, the H+/e- ratio approached 2 as a limit for both acceptors. A working model is presented which includes two sites of proton translocation, one between the photoacts, the other connected to Photosystem I, each of which translocates two protons per electron. Each site presents a approximately 30 ms diffusion barrier to proton passage which can be lowered by uncouplers to 6-10 ms.

Chloroplasts↗
PubMed1972

Photon trapping in photosystem II of photosynthesis. The fluorescence rise curve in the presence of 3-(3,4-dichlorophenyl)-1,1-dimetnhylurea.

Using isolated chloroplasts in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), an analysis was made of the rise of the fluorescence yield effected by weak light. Depending on the pretreatment, the time-course of the rapid photochemical part of the rise varied between nearly first-order and quadratic kinetics, i.e., reflected either a one-quantum or a two-quantum conversion. We consider the occurrence of two photoreductants per system II unit, which are reoxidized in different dark reactions. The data further showed that the "first-order process" is also inhomogeneous.

Chloroplasts↗