Proton efflux from oat coleoptile cells and exchange with wall calcium after IAA or fusicoccin treatment

Elongation growth of plant cells occurs by stretching of cell walls under turgor pressure when intermolecular bonds in the walls are temporarily loosened. The acid-growth theory predicts that wall loosening is the result of wall acidification because treatments (including IAA and fusicoccin) that cause lowered wall pH cause elongation. However, conclusive evidence that IAA primarily reduces wall pH has been lacking. Calcium has been reported to stiffen the cell walls. We have used a microelectrode ion-flux measuring technique to observe directly, and non-invasively, the net fluxes of protons and calcium from split coleoptiles of oats (Avena sativa L.) in unbuffered solution. Normal net fluxes are 10 nmol · m⁻² · s⁻¹ proton efflux and zero calcium flux. The toxin fusicoccin (1 μM) causes immediate efflux from tissue not only of protons, but also of calcium, about 110 nmol · m⁻² · s⁻¹ in each case. The data fit the “weak acid Donnan Manning” model for ion exchange in the cell wall. Thus we associate the known “acid-growth” effect of fusicoccin with the displacement of calcium from the wall by exchange for protons extruded from the cytoplasm. Application of 10 μM IAA causes proton efflux to increase transiently by about 15 nmol · m⁻² · s⁻¹ with a lag of about 10 min. The calcium influx decreases immediately to an efflux of about 20 nmol · m⁻² · s⁻¹. It appears that auxin too causes an “acid-growth” effect, with extruded protons exchanging for calcium in the cell walls. I. Arif is currently recieving an AIDAB scholarship. This work was supported by an Australian Research Council grant to I.A. Newman.     

Comparative analysis of four <Emphasis Type="Italic">β</Emphasis>-galactosidases from <Emphasis Type="Italic">Bifidobacterium bifidum</Emphasis> NCIMB41171: purification and biochemical characterisation

Four different β-galactosidases (previously named BbgI, BbgII, BbgIII and BbgIV) from Bifidobacterium bifidum NCIMB41171 were overexpressed in Escherichia coli, purified to homogeneity and their biochemical properties and substrate preferences comparatively analysed. BbgI was forming a hexameric protein complex of 875 kDa, whereas BbgII, BbgIII and BbgIV were dimers with native molecular masses of 178, 351 and 248 kDa, respectively. BbgII was the only enzyme that preferred acidic conditions for optimal activity (pH 5.4–5.8), whereas the other three exhibited optima in more neutral pH ranges (pH 6.4–6.8). Na⁺ and/or K⁺ ions were prerequisite for BbgI and BbgIV activity in Bis–Tris-buffered solutions, whereas Mg⁺⁺ was strongly activating them in phosphate-buffered solutions. BbgII and BbgIII were slightly influenced from the presence or absence of cations, with Mg⁺⁺, Mn⁺⁺ and Ca⁺⁺ ions exerting the most positive effect. Determination of the specificity constants (k _cat/K _m) clearly indicated that BbgI (6.11 × 10⁴ s⁻¹ M⁻¹), BbgIII (2.36 × 10⁴ s⁻¹ M⁻¹) and especially BbgIV (4.01 × 10⁵ s⁻¹ M⁻¹) are highly specialised in the hydrolysis of lactose, whereas BbgII is more specific for β-d-(1→6) galactobiose (5.59 × 10⁴ s⁻¹ M⁻¹) than lactose (1.48 × 10³ s⁻¹ M⁻¹). Activity measurements towards other substrates (e.g. β-d-(1→6) galactobiose, β-d-(1→4) galactobiose, β-d-(1→4) galactosyllactose, N-acetyllactosamine, etc.) indicated that the β-galactosidases were complementary to each other by hydrolysing different substrates and thus contributing in a different way to the bacterial physiology.     

Characterisation of acidic sites of Pseudomonas biomass capable of binding protons and cadmium and removal of cadmium via biosorption

Summary The ability of Pseudomonas aeruginosa to accumulate Cd(II) ions from wastewater industries was experimentally investigated and mathematically modelled. From the potentiometric titration and non-ideal competitive analysis (NICA) model, it was found that the biomass contains three acidic sites. The values of proton binding (pK _i =1.66±3.26×10⁻³, 1.92±1.63×10⁻⁴ and 2.16±3.79×10⁻⁴) and binding constant of cadmium metal ions (pK _M1=1.99±2.45×10⁻³ and pK _M2=1.67±4.08×10⁻³) on the whole surface of biomass showed that protonated functional groups and biosorption of Cd(II) ions could be attributed to a monodentate binding to one acidic site, mainly the carboxylic group. From the isothermal sorption experimental data and Langmuir model, it was also found that the value of Langmuir equilibrium (pK _f) constant is 2.04±2.1×10⁻⁵ suggesting that the carboxyl group is the main active binding site. In addition, results showed that the maximum cadmium capacity (q _max) and affinity of biomass towards cadmium metal ions (b) at pH 5.1 and 20 min were 96.5±0.06 mg/g and 3.40×10⁻³± 2.10×10⁻³, respectively. Finally, interfering metal ions such as Pb(II), Cu(II), Cr(III), Zn(II), Fe(II), Mn(II), Ca(II) and Mg(II) inhibited Cd(II) uptake. Comparing the biosorption of Cd(II) by various Pseudomonas isolates from contaminated environment samples (soil and sewage treatment plant) showed that maximum capacities and equilibrium times were different, indicating that there was a discrepancy in the chemical composition between biomasses of different strains.     

Dynamics of Calcium Fluxes in Nonexcitable Cells: Mathematical Modeling

Mathematical models simulating the dynamics of calcium redistribution (elicited by experimental interference with the pathways of calcium fluxes) in cellular compartments have been developed, based on a minimal scheme of the pathways of calcium fluxes in nonexcitable cells suspended in calcium-free medium. The models are consistent with available experimental data. All parameters are quantitatively related to the intrinsic properties of calcium adenosine triphosphatases (ATPases) and cellular membranes; there is no interdependence between the parameters. The models can be used as the basis for quantitative analysis and interpretation of experimental data. The activities of plasma membrane and sarcoendoplasmic reticulum calcium ATPases (PMCA and SERCAs) are governed by different mechanisms. PMCA is likely to undergo transitions from inactive to active to “dormant” (not identical to the initial) and back to inactive states, the mean duration of the cycle lasting for minutes or longer. The sequence of the transitions is initiated, presumably, by an increase in cytosolic calcium concentration. The transition of PMCA from inactive to active (at least at low rates of increase in cytosolic calcium concentration) is likely to be slower than that from active to dormant. SERCA, presumably, transits from inactive to active state in response to increases in calcium leakage from calcium stores. Whereas PMCA extrudes excess calcium (a definite quantity of it) in a short pulse, SERCA retakes calcium back into the stores permanently at a high rate. The models presented here may be the best means for the moment to quantitatively relate the dynamics of calcium fluxes in nonexcitable cells with known or putative properties of the mechanisms underlying activation of calcium ATPases.     

Indole-3-Acetic Acid Control on Acidic Oat Cell Wall Peroxidases

Incubation of oat coleoptile segments with 40 μm indoleacetic acid (IAA) induced a decrease of 35–60% in peroxidase activity at the cell wall compartment. Treatment with IAA also produced a similar decrease in the oxidation of NADH and IAA at the cell wall. Isoelectric focusing of ionic, covalent, and intercellular wall peroxidase fractions showed that acidic isoforms (pI 4.0–5.5) were reduced preferentially by IAA treatment. Marked differences were found between acidic and basic wall isoperoxidases in relation to their efficacy in the oxidation of IAA. A peroxidase fraction containing acidic isoforms oxidized IAA with a V _max/s_0.5 value of 2.4 × 10⁻² min⁻¹· g fw⁻¹, 4.0 times higher than that obtained for basic peroxidase isoforms (0.6 × 10⁻² min⁻¹· g fw⁻¹). In contrast, basic isoforms were more efficient than acidic isoperoxidases in the oxidation of coniferyl alcohol or ferulic acid with H₂O₂ (5.6 and 2.1 times, respectively). The levels of diferulate and lignin in the walls of oat coleoptile segments were not altered by treatment with IAA. The decrease in cell wall peroxidase activity by IAA was related more to reduced oxidative degradation of the hormone than to covalent cell wall cross-linking. Received November 1, 1998; accepted December 14, 1998     

Light-induced Mn2+ influx in Limulus ventral photoreceptors

In contrast to insect species, light-activated influx of divalent ions into Limulus ventral photoreceptors has proven difficult to demonstrate. We used the quench of the fluorescent indicator dye, fura-2, to measure Mn²⁺ influx. Limulus ventral photoreceptors were injected with fura-2 and excited at 360 nm. When the photoreceptors were bathed in 1 mmol · l⁻¹ Mn²⁺, an approximately 1% per 10 s decline in the fura-2 fluorescence during intervals between 50-ms flashes was taken as a measure of Mn²⁺ entry in darkness. Fluorescence decline during 10-s flashes was used to monitor Mn²⁺ entry during the photoresponse. During the 10-s flashes we observed a small rapid decline of the fura-2 fluorescence even in the absence of Mn²⁺. This reflected a contamination of the fluorescence signal arising from light-induced release of intracellular calcium stores. A subsequent slower decline in fluorescence during the 10-s flash, amounting to approximately 9% per 10 s, was only observed in the presence of extracellular Mn²⁺ and was attributed to Mn²⁺ influx. This light-activated influx was not through voltage-gated calcium channels since it persisted under voltage clamp, was not stimulated by depolarizing current injections, nor blocked by NiCl₂. Depletion of internal calcium stores by cyclopiazonic acid treatment did not accelerate Mn²⁺ influx. Accepted: 30 January 1998     

Kinetics of N-substituted phenothiazines and N-substituted phenoxazines oxidation catalyzed by fungal laccases

Laccase-catalyzed oxidation of N-substituted phenothiazines and N-substituted phenoxazines was investigated at pH 5.5 and 25°C. The recombinant laccase from Polyporus pinsitus (rPpL) and the laccase from Myceliophthora thermophila (rMtL) were used. The dependence of initial reaction rate on substrate concentration was analyzed by applying the laccase action scheme in which the laccase native intermediate (NI) reacts with a substrate forming reduced enzyme. The reduced laccase produces peroxide intermediate (PI) which in turn decays to the NI. The calculated constant (k_ox) values of the PI formation are (6.1±3.1)×10⁵ M⁻¹s⁻¹ for rPpL and (2.5±0.9)×10⁴ M⁻¹s⁻¹ for rMtL. The bimolecular constants of the reaction of the native intermediate with electron donor (kred) vary in the interval from 2.2×10⁵ to 2.1×10⁷ M⁻¹s⁻¹ for rPpL and from 1.3×10² to 1.8×10⁵ M^-1s⁻¹ for rMtL. The larger reactivity of rPpL in comparison to rMtL is associated with the higher redox potential of type I Cu of rPpL. The variation of k_red values for both laccases correlates with the change of the redox potential of substrates. Following outer sphere (Marcus) electron transfer mechanism the calculated activationless electron transfer rate and the apparent reorganization energy are 5.0×107 M⁻¹s⁻¹ and 0.29 eV, respectively.     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000     

Polyanion-mediated mineralization — mineralization in coccolithophore (Pleurochrysis carterae) variants which do not express PS2, the most abundant and acidic mineral-associated polyanion in wild-type cells

Summary Highly acidic macromolecules are postulated intermediates in biomineralization because they sequester large numbers of calcium ions and occur in high concentrations at mineralizing foci in distantly related organisms. The mineral-associated polyanions in the coccolithophorePleurochrysis carterae are acidic polysaccharides which surround calcite elements on mineralized scales called coccoliths. PS2 is the most abundant and acidicPleurochrysis polyanion, containing four carboxyl groups per disaccharide repeat. This study examines the mineralizing function of PS2 by analysis of calcium carbonate deposition in PS2-deficient cells. Mineral-deficientPleurochrysis variants were separated from wild-type cells by density gradient centrifugation and analyzed for PS2 expression. Three independent low-density variants (two spontaneous and one chemically-induced) not expressing PS2 (ps2⁻) were isolated. The mineral content of each ps2⁻ variant was less than 5% of the wild-type level. Most ps2⁻ coccoliths were unmineralized, but when crystals were present, they occurred on the coccolith rim as in wild-type cells. When ps2⁻ rim elements were adequately large and numerous, they displayed the alternating A and B anvil shapes characteristic of wild-type rim elements. Although ps2⁻ cells may have defects which affect the expression of cellular components in addition to PS2, correlation of PS2 nonexpression with mineral deficiency in three independent variants supports the hypothesis that PS2 is a functional intermediate in mineral deposition. PS2 may affect the level ofPleurochrysis mineralization by (a) controlling the amount of calcium ions available for mineralization, (b) accelerating the rate of calcite nucleation, or (c) both. However, PS2 does not influence crystal morphology, since the more mature crystals in ps2⁻ cells exhibit the anvil-shaped morphology of wild-type crystals.     

Toxicity assessment of paraverine hydrochloride and papaverine-derived metabolites in primary cultures of rat hepatocytes

Summary The present study was undertaken to assess and compare the toxic effects of papaverine hydrochloride and its metabolites. Primary cell cultures of rat hepatocytes were treated with papavarine (papaver), 3′-O-desmethyl (3′-OH), 4′-O-desmethyl (4′-OH), and 6-O-desmethyl (6-OH) papaverine at 1×10⁻⁵, 1×10⁻⁴, and 1×10⁻³ M for 4,8, 12, and 24-h periods. Cell injury was determined by: a) cell viability using the trypan blue exclusion test; b) cytosolic enzyme leakage of lactate dehydrogenase and aspartate aminotransferase; c) morphologic alterations; and d) lactate: pyruvate (L:P) ratios. Cell cultures showed concentration-and time-dependent responses. For example, a decrease in cell viability and an increase in enzyme leakage were observed after cell treatment with 1×10⁻⁴ and 1×10⁻³ M papaver for 8 h; 1×10⁻³ M 6-OH papaverine for 8 h and 1×10⁻⁴ M for 24 h; and 1×10⁻³ M 4′-OH papaverine for 24 h (P<0.05). Furthermore, changes in morphology correlated to cell viability and enzyme release in those cultures treated with papaver, 4′-OH and 6-OH papaverine. Some of these changes included size deformation, cell detachment from the dishes, and cell necrosis. On the other hand, an increase in L:P ratios (P<0.05) was detected with papaver as early as 8 h with 1×10⁻⁴ and 1×10⁻³ M and 12 h with 1×10⁻⁵ M; 6-OH showed an increase, in L:P ratios at 8 h with 1×10⁻³ M and 12 h with 1×10⁻⁴ M; these changes were evident with 4′-OH at 12 h with 1×10⁻³ M. In contrast, cells treated with 3′-OH papaverine did not show significant damage with any time period and concentration used in this study. The results of this study indicate that papaverine-derived metabolites are less cytotoxic than its parent compound, papaver. The toxicity was ranked as follows: papaver>6-OH>4′-OH>−3′-OH. This work was supported in part by grant ES04200-02 from the National Institute of Environmental Health Sciences, Bethesda, MD. Presented in part at the fall ASPET meeting in Salt Lake City, August, 1989. Daniel Acosta is a Burroughs Wellcome Scholar in Toxicology.     

The influence of differentiation-inducing agents on plasma membrane surface characteristics of THP-1 cells

Undifferentiated THP-1 cells from Cell Culture Collection of the Institute of Cytology, RAS (St. Petersburg), are characterized by weak expression of Toll-like receptor-4 (TLR4) on the cell surface (up to 2%) and by almost undetectable expression of CD14 and CD11b receptors. Differentiation agent phorbol-12-myristate-13-acetate independently of its concentration (2 × 10⁻⁷ M or 10⁻⁸ M) and incubation time (24 or 48 h) did not initiate CD11b surface expression and did not change the parameter S_app (0.605 ± 0.005 at 37°C) reflecting the cell membrane viscosity. Differentiation of THP-1 cells induced by another differentiation agent, 1α,25-dihydroxyvitamin D₃, caused expression of CD14 (up to 70–80%) and CD11b (up to 15–20%) receptors, again without changes in plasma membrane viscosity. The rate constants of the reduction of 5- and 16-doxyl-stearic acids by THP-1 cells were in the range of 6–8 × 10⁻³ s⁻¹ at 37°C. During cell differentiation significant changes in cell electrophoretic mobility (EM, μm s⁻¹ V⁻¹ cm) were observed. Mean value of EM for undifferentiated THP-1 cells was −1.332 ± 0.011, whereas for phorbol-12-myristate-13-acetate- and 1α,25-dihydroxyvitamin D₃-treated cells it was −1.432 ± 0.030 and −1.212 ± 0.016, respectively.     

Laser flash photolysis study on antioxidant properties of hydroxycinnamic acid derivatives

The antioxidant properties of hydroxycinnamic acid derivatives (HCA) were studied by laser flash photolysis. The transient species with maximum absorption at 360 nm were assigned to the phenoxyl radical of HCA. The SO₄^•− induced oxidation of HCA was also investigated. It was shown that the interaction of SO₄^•− with HCA resulted in the formation of HCA phenoxyl radicals with rate constants of 2.0–3.9×10⁹ M⁻¹ s⁻¹. The reactions of HCA with triplet state of benzophenone were analyzed and quenching rate constants of 4.3–7.8×10⁹ M⁻¹ s⁻¹ were determined.     

Studies on water transport through the sweet cherry fruit surface: characterizing conductance of the cuticular membrane using pericarp segments

Water conductance of the cuticular membrane (CM) of mature sweet cherry fruit (Prunus avium L. cv. Sam) was investigated by monitoring water loss from segments of the outer pericarp excised from the cheek of the fruit. Segments consisted of epidermis, hypodermis and several cell layers of the mesocarp. Segments were mounted in stainless-steel diffusion cells with the mesocarp surface in contact with water, while the outer cuticular surface was exposed to dry silica (22 ± 1 °C). Conductance was calculated by dividing the amount of water transpired per unit area and time by the difference in water vapour concentration across the segment. Conductance values had a log normal distribution with a median of 1.15 × 10⁻⁴ m s⁻¹ (n=357). Transpiration increased linearly with time. Conductance remained constant and was not affected by metabolic inhibitors (1 mM NaN₃ or 0.1 mM carbonylcyanide m-chlorophenylhydrazone) or thickness of segments (range 0.8–2.8 mm). Storing fruit (up to 42 d, 1 °C) used as a source of segments had no consistent effect on conductance. Conductance of the CM increased from cheek (1.16 ± 0.10 × 10⁻⁴ m s⁻¹) to ventral suture (1.32 ± 0.07 × 10⁻⁴ m s⁻¹) and to stylar end (2.53 ± 0.17 × 10⁻⁴ m s⁻¹). There was a positive relationship (r²=0.066**; n=108) between conductance and stomatal density. From this relationship the cuticular conductance of a hypothetical astomatous CM was estimated to be 0.97 ± 0.09 × 10⁻⁴ m s⁻¹. Removal of epicuticular wax by stripping with cellulose acetate or extracting epicuticular plus cuticular wax by dipping in CHCl₃/methanol increased conductance 3.6- and 48.6-fold, respectively. Water fluxes increased with increasing temperature (range 10–39 °C) and energies of activation, calculated for the temperature range from 10 to 30 °C, were 64.8 ± 5.8 and 22.2 ± 5.0 kJ mol⁻¹ for flux and vapour-concentration-based conductance, respectively. Received: 23 March 2000 / Accepted: 28 July 2000     

Olive mill wastewater disposal in evaporation ponds in Sfax (Tunisia): moisture content effect on microbiological and physical chemical parameters

The study of the isotherms desorption of olive mill wastewater (OMW) was investigated to describe its water activity under different saturated environments. The microbial biodegradation of OMW during its storage in 5 evaporation ponds located in Agareb (Sfax-Tunisia) was carried out during the oil-harvesting year held 105 days in 2004. Gravimetric static method using saturated salt solutions was used and OMW as placed at 30°C and under different water activities ranging from 0.11 to 0.90. Eight models were taken from the literature to describe experimental desorption isotherms. During storage, the evolution of physico-chemical parameters including pH, temperature, evaporation, humidity, total phosphorus, chemical oxygen demand (COD), biological oxygen demand (BOD) and phenols and three microbiological flora (aerobic mesophilic bacteria, yeasts and moulds) were considered. At 30°C, when relative humidity increased in the experimented ponds of 69, 84 and 90%, the evaporation speed decreased from 1.24 × 10⁻⁵ to 5 × 10⁻⁶ cm³ s⁻¹, from 6 × 10⁻⁵ to 7 × 10⁻⁶ cm³ s⁻¹ and from 5 × 10⁻⁶ to 1.1 × 10⁻⁷ cm³ s⁻¹ respectively. The desorption isotherm exhibited a sigmoidal curve corresponding to type II, typical of many organic material. The GAB and Peleg models gave the best fit for describing the relationship between the equilibrium moisture content and water activity in OMW (R ² = 0.998). During the storage period, the analysis showed an increase of all the physico-chemical parameters studied, except phenols and total phosphorus concentrations. The microbiological study showed the predominance of yeasts and moulds and the decrease of bacteria population after 75 days reflecting both effect of recalcitrant compounds and the water activity on microbial growth.     

Resistance to glyphosate in the cyanobacterium <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> as result of pre-selective mutations

Adaptation of Microcystis aeruginosa (Cyanobacteria) to resist the herbicide glyphosate was analysed by using an experimental model. Growth of wild-type, glyphosate-sensitive (G^s) cells was inhibited when they were cultured with 120 ppm glyphosate, but after further incubation for several weeks, occasionally the growth of rare cells resistant (G^r) to the herbicide was found. A fluctuation analysis was carried out to distinguish between resistant cells arising from rare spontaneous mutations and resistant cells arising from other mechanisms of adaptation. Resistant cells arose by rare spontaneous mutations prior to the addition of glyphosate, with a rate ranging from 3.1 × 10⁻⁷ to 3.6 × 10⁻⁷ mutants per cell per generation in two strains of M. aeruginosa; the frequency of the G^r allele ranged from 6.14 × 10⁻⁴ to 6.54 × 10⁻⁴. The G^r mutants are slightly elliptical in outline, whereas the G^s cells are spherical. Since G^r mutants have a diminished growth rate, they may be maintained in uncontaminated waters as the result of a balance between new resistants arising from spontaneous mutation and resistants eliminated by natural selection. Thus, rare spontaneous pre-selective mutations may allow the survival of M. aeruginosa in glyphosate-polluted waters via G^r clone selection.     

Internal dose assessment of <Superscript>210</Superscript>Po using biokinetic modeling and urinary excretion measurement

The mysterious death of Mr. Alexander Litvinenko who was most possibly poisoned by Polonium-210 (²¹⁰Po) in November 2006 in London attracted the attention of the public to the kinetics, dosimetry and the risk of this high radiotoxic isotope in the human body. In the present paper, the urinary excretion of seven persons who were possibly exposed to traces of ²¹⁰Po was monitored. The values measured in the GSF Radioanalytical Laboratory are in the range of natural background concentration. To assess the effective dose received by those persons, the time-dependence of the organ equivalent dose and the effective dose after acute ingestion and inhalation of ²¹⁰Po were calculated using the biokinetic model for polonium (Po) recommended by the International Commission on Radiological Protection (ICRP) and the one recently published by Leggett and Eckerman (L&E). The daily urinary excretion to effective dose conversion factors for ingestion and inhalation were evaluated based on the ICRP and L&E models for members of the public. The ingestion (inhalation) effective dose per unit intake integrated over one day is 1.7 × 10⁻⁸ (1.4 × 10⁻⁷) Sv Bq⁻¹, 2.0 × 10⁻⁷ (9.6 × 10⁻⁷) Sv Bq⁻¹ over 10 days, 5.2 × 10⁻⁷ (2.0 × 10⁻⁶) Sv Bq⁻¹ over 30 days and 1.0 × 10⁻⁶ (3.0 × 10⁻⁶) Sv Bq⁻¹ over 100 days. The daily urinary excretions after acute ingestion (inhalation) of 1 Bq of ²¹⁰Po are 1.1 × 10⁻³ (1.0 × 10⁻⁴) on day 1, 2.0 × 10⁻³ (1.9 × 10⁻⁴) on day 10, 1.3 × 10⁻³ (1.7 × 10⁻⁴) on day 30 and 3.6 × 10⁻⁴ (8.3 × 10⁻⁵) Bq d⁻¹ on day 100, respectively. The resulting committed effective doses range from 2.1 × 10⁻³ to 1.7 × 10⁻² mSv by an assumption of ingestion and from 5.5 × 10⁻² to 4.5 × 10⁻¹ mSv by inhalation. For the case of Mr. Litvinenko, the mean organ absorbed dose as a function of time was calculated using both the above stated models. The red bone marrow, the kidneys and the liver were considered as the critical organs. Assuming a value of lethal absorbed dose of 5 Gy to the bone marrow, 6 Gy to the kidneys and 8 Gy to the liver, the amount of ²¹⁰Po which Mr. Litvinenko might have ingested is therefore estimated to range from 27 to 1,408 MBq, i.e 0.2–8.5 μg, depending on the modality of intake and on different assumptions about blood absorption.     

Kinetics of isomerization for the proline helical forms of two oligoproline redox arrays by circular dichroic spectropolarimetry

Summary The kinetics of isomerization of the helical forms of three oligoprolines was determined by far-ultraviolet CD spectropolarimetry and kinetic analysis by singular value decomposition. ZRA (Pro₃-X-Pro₂-Y-Pro₂-Z-Pro₃) and ZRA2 (Pro₇-X-Pro₂-Y-Pro₂-Z-Pro₇) bear large redox-active substituents on proline residues X, Y, and Z, but P9 (Pro₉) does not. All three peptides formed a stable proline-II helix in water. In acetonitrile, both ZRA2 and P9 were converted into a proline-I helical form but ZRA remained predominantly in the proline-II helical form. Evidently, in order to undergo substantial proline II→I isomerization, an oligoproline chain containing large substituents needs to have a segment of consecutive unsubstituted proline residues that is sufficiently long to form a stable proline helix. Biexponential kinetics (A→B, k₁=∼3.3×10⁻⁴s⁻¹; B→C, k₂=∼0.8×10⁻⁴s⁻¹) were observed for the proline II→I isomerization of ZRA2 and P9 in acetonitrile and for the proline I→II isomerization of ZRA2 in water, which provides evidence for the growth and decay of a major kinetic intermediate.     

Swimming behaviour of <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> in relation to food concentration and feeding rates

We examined feeding rates and swimming speed in amictic females of Brachionus plicatilis over algal cell concentrations ranging from 15 × 10³ to 30 × 10⁶ cell ml⁻¹, to determine to what extent filtration rate is a consequence of a real modulation of swimming speed in response to food availability. Swimming rates were measured using an automated motion analysis system via video recording. The results showed that swimming speed changed as a function of food density. Swimming speed increased from the lowest tested concentration of algae to reach a maximum at 6 × 10⁶ cell ml⁻¹. Above this density, swimming speed declined slightly and then remained constant at a mean speed of 0.45 mm s⁻¹. Filtration and ingestion rates changed as cell concentration increased, following patterns consistent with those generally described for suspension feeders. However, the observed swimming pattern did not explain the recorded changes in clearance rate. These results suggest that filtration, and therefore ingestion, is mainly regulated by modifying particle retention efficiency. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer Research     

The effect of organic toxicants on sensitivity of intestinal glycosidases to Cu and Zn in juvenile roach

The chronic effect (duration of exposure 218 days) of polychlorinated biphenyls (PCBs) and the prolonged effect of the short-term action of chlorophos or of N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) during embryogenesis upon the sensitivity of intestinal glycosidases to Cu and Zn was studied in roach (Rutilus rutilus (L.) underyearlings. The Cu⁺² and Zn⁺² ions at concentrations ranging from 0.1 to 25 mg/L in vitro cause a 10–77% decrease in amylolytic activity in the intestinal mucosa of control roach. An elevated level of PCBs (50.8 ng/g wet weight of food and 426 ng/g dry weight of ground) increased the sensitivity of glycosidases to Cu and Zn. The embryotoxic action of chlorophos at concentrations of 1 × 10⁻⁶−1 × 10⁻² mg/L in most cases increased the inhibitory effect of Cu but decreased that of Zn. As a rule, MNNG (3 × 10⁻⁷−3 × 10⁻¹ mg/L) reduced the glycosidase sensitivity to the effect of metal ions. The magnitude and direction of the effect depend on the nature and concentration of toxicants.