Resistance to CO2 diffusion in cuticular membranes of amphibious plants and the implication for CO2 acquisition

Cuticular membranes (CMs) were isolated from leaves of amphibious and submerged plants and their CO2 resistances were determined as a contribution to establish quantitatively the series of resistances met by CO2 diffusing from bulk water to the chloroplasts of submerged leaves. The isolation was performed enzymatically; permeabilities were determined and converted to resistances. The range of permeance values was 3 to 43 x 10(-6) m s(-1) corresponding to resistance values of 23 to 295 x 10(3) s m(-1), i.e. of the same order of magnitude as boundary layer resistances. The sum of boundary layer, CM, leaf cell and carboxylation resistances could be contained within the total diffusion resistance as determined from the photosynthetic CO2 affinity of the leaf. From the same species, the aerial leaf CM resistance was always higher than the aquatic leaf CM resistance. In a terrestrial plant, the CM resistance to CO2 diffusion was found lower in leaves developed submerged.     

Water permeability of isolated cuticular membranes: The effect of cuticular waxes on diffusion of water

Summary The water permeability of astomatous cuticular membranes isolated from Citrus aurantium L. leaves, pear (Pyrus communis L.) leaves and onion (Allium cepa L.) bulb scales was determined before and after extraction of cuticular waxes with lipid solvents. In pear, the permeability coefficients for diffusion of tritiated water across cuticular membranes (CM) prior to extraction [P _d(CM)] decreased by a factor of four during leaf expansion. In all three species investigated P _d(CM) values of cuticular membranes from fully expanded leaves varied between 1 to 2×10^-7 cm^-3 s^-1·P _d(CM) values were not affected by pH. Extraction of cuticular waxes from the membranes increased their water permeability by a factor of 300 to 500. Permeability coefficients for diffusion of THO across the cutin matrix (MX) after extraction [P _d(MX)] increased with increasing pH. P _dvalues were not inversely proportional to the thickness of cuticular membranes. By treating the cutin matrix and cuticular waxes as two resistances acting in series it was shown that the water permeability of cuticles is completely determined by the waxes. The lack of the P _d(CM) values to respond to pH appeared to be due to structural effects of waxes in the cutin matrix. Cuticular membranes from the submerse leaves of the aquatic plant Potamogeton lucens L. were three orders of magnitude more permeable to water than the cuticular membranes of the terrestrial species investigated.Abbreviations CM cuticular membrane - MX cutin matrix - WAX waxes This study was supported by a grant from the Deutsche Forschungsgemeinschaft.     

Partitioning of mesophyll conductance for CO2 into intercellular and cellular components using carbon isotope composition of cuticles from opposite leaf sides

We suggest a new technique for estimating the relative drawdown of CO₂ concentration (c) in the intercellular air space (IAS) across hypostomatous leaves (expressed as the ratio c_d/c_b, where the indexes d and b denote the adaxial and abaxial edges, respectively, of IAS), based on the carbon isotope composition (δ¹³C) of leaf cuticular membranes (CMs), cuticular waxes (WXs) or epicuticular waxes (EWXs) isolated from opposite leaf sides. The relative drawdown in the intracellular liquid phase (i.e., the ratio c_c/c_bd, where c_c and c_bd stand for mean CO₂ concentrations in chloroplasts and in the IAS), the fraction of intercellular resistance in the total mesophyll resistance (r_IAS/r_m), leaf thickness, and leaf mass per area (LMA) were also assessed. We show in a conceptual model that the upper (adaxial) side of a hypostomatous leaf should be enriched in ¹³C compared to the lower (abaxial) side. CM, WX, and/or EWX isolated from 40 hypostomatous C₃ species were ¹³C depleted relative to bulk leaf tissue by 2.01–2.85‰. The difference in δ¹³C between the abaxial and adaxial leaf sides (δ¹³C_AB ? ¹³C_AD, Δ_b–d), ranged from ??2.22 to +?0.71‰ (??0.09 ± 0.54‰, mean ± SD) in CM and from ??7.95 to 0.89‰ (??1.17 ± 1.40‰) in WX. In contrast, two tested amphistomatous species showed no significant Δ_b–d difference in WX. Δ_b–d correlated negatively with LMA and leaf thickness of hypostomatous leaves, which indicates that the mesophyll air space imposes a non-negligible resistance to CO₂ diffusion. δ¹³C of EWX and 30-C aldehyde in WX reveal a stronger CO₂ drawdown than bulk WX or CM. Mean values of c_d/c_b and c_c/c_bd were 0.90 ± 0.12 and 0.66 ± 0.11, respectively, across 14 investigated species in which wax was isolated and analyzed. The diffusion resistance of IAS contributed 20 ± 14% to total mesophyll resistance and reflects species-specific and environmentally-induced differences in leaf functional anatomy.

     

The 'hydrology' of leaves: co-ordination of structure and function in temperate woody species

The hydraulic conductance of the leaf lamina (K_lamina) substantially constrains whole‐plant water transport, but little is known of its association with leaf structure and function. K_lamina was measured for sun and shade leaves of six woody temperate species growing in moist soil, and tested for correlation with the prevailing leaf irradiance, and with 22 other leaf traits. K_lamina varied from 7.40 × 10^?5 kg m^?2 s^?1 MPa^?1 for Acer saccharum shade leaves to 2.89 × 10^?4 kg m^?2 s^?1 MPa^?1 for Vitis labrusca sun leaves. Tree sun leaves had 15–67% higher K_lamina than shade leaves. K_lamina was co‐ordinated with traits associated with high water flux, including leaf irradiance, petiole hydraulic conductance, guard cell length, and stomatal pore area per lamina area. K_lamina was also co‐ordinated with lamina thickness, water storage capacitance, 1/mesophyll water transfer resistance, and, in five of the six species, with lamina perimeter/area. However, for the six species, K_lamina was independent of inter‐related leaf traits including leaf dry mass per area, density, modulus of elasticity, osmotic potential, and cuticular conductance. K_lamina was thus co‐ordinated with structural and functional traits relating to liquid‐phase water transport and to maximum rates of gas exchange, but independent of other traits relating to drought tolerance and to aspects of carbon economy.     

Gas permeability of plant cuticles

Cuticles from the adaxial surface of Citrus aurantium L. leaves and from the pericarp of Lycopersicon esculentum L. and Capsicum annuum L. were isolated enzymatically and their oxygen permeability was determined. Isolated cuticles were mounted between a gaseous and an aqueous compartment with the physiological outer side of the membrane facing the gaseous compartment. Permeability for oxygen was characterized by permeability (P) and diffusion (D) coefficients. P and D were independent of the driving force (gradient of oxygen concentration) across the cuticle, thus, Henry's law was obeyed. P values for the diffusion of oxygen varied between 3·10^-7 (Citrus), 1.4·10^-6 (Capsicum), and 1.1·10^-6 (Lycopersicon) m·s^-1. Extraction of soluble lipids from the cuticles increased the permeability. By treating the cutin matrix and the soluble lipids as resistances in series, it could be demonstrated that the soluble lipids were the main resistance for oxygen permeability in Citrus cuticles. However, in Lycopersicon and Capsicum, both the cutin matrix and the soluble lipids determined the total resistance. P values were not affected by either the proton concentration (pH 3–9) or the cations (Na⁺, Ca²⁺) present at the morphological inner side of the cuticles. It is concluded that the water content of cuticles does not affect the permeability properties for oxygen. Partition coefficients indicated a high solubility of oxygen in the cuticle of Citrus. The data suggest a solubility process in the cuticle of Citrus with respect to oxygen permeation.Abbreviations CM cuticular membrane - MX cutin polymer matrix - SCL soluble cuticular lipids     

THE RELATIONSHIP BETWEEN THE THICKNESS OF DECIDUOUS LEAVES AND THEIR MAXIMUM PHOTOSYNTHETIC RATE

Mc Clendon , John H. (U. Delaware, Newark.) The relationship between the thickness of deciduous leaves and their maximum photosynthetie rate. Amer. Jour. Bot. 49(4): 320–322. Illus. 1962.—Data of Willstätter and Stoll (25°, saturating light and CO₂) are used to show that it is useful to plot photosynthetie rate per unit area as a function of the density thickness (g/cm²; fresh wt) of the leaves. Data for 23 species were plotted together. If P is photosynthetie rate, T the total density thickness, E the density thickness of the epidermis, a linear relation is found using the maximum values of P, such that P = R (T - E), where R is about 30 (mg CO₂) (hr)^-1 (g F.W.)^-1. Using this expression, 15 species were represented by values of R over 23, while very young leaves, aurea² leaves and a few others fell in the range 8–16. A mean value of E was taken, from other data, to be about 3 × 10^-3 g/cm², while T ranged from 10 to 40 × 10^-3 g/cm², and P ranged from 0.2 to 0.8 (mg CO₂) hr^-1 cm^-2 for “normal” leaves.     

Carbon: freshwater plants

δ¹³C values for freshwater aquatic plant matter varies from ?11 to ?50‰ and is not a clear indicator of photosynthetic pathway as in terrestrial plants. Several factors affect δ¹³C of aquatic plant matter. These include: (1) The δ¹³C signature of the source carbon has been observed to range from +1‰ for HCO₃^? derived from limestone to ?30‰ for CO₂ derived from respiration. (2) Some plants assimilate HCO₃^?, which is –7 to –11‰ less negative than CO₂. (3) C₃, C₄, and CAM photosynthetic pathways are present in aquatic plants. (4) Diffusional resistances are orders of magnitude greater in the aquatic environment than in the aerial environment. The greater viscosity of water acts to reduce mixing of the carbon pool in the boundary layer with that of the bulk solution. In effect, many aquatic plants draw from a finite carbon pool, and as in terrestrial plants growing in a closed system, biochemical discrimination is reduced. In standing water, this factor results in most aquatic plants having a δ¹³C value similar to the source carbon. Using Farquhar's equation and other physiological data, it is possible to use δ¹³C values to evaluate various parameters affecting photosynthesis, such as limitations imposed by CO₂ diffusion and carbon source.     

Comparative NMR studies of diffusional water permeability of red blood cells from different species: XVIII platypus (Ornithorhynchus anatinus) and saltwater crocodile (Crocodylus porosus)

As part of a programme of comparative measurements of P _d (diffusional water permeability) the RBCs (red blood cells) from an aquatic monotreme, platypus (Ornithorhynchus anatinus), and an aquatic reptile, saltwater crocodile (Crocodylus porosus) were studied. The mean diameter of platypus RBCs was estimated by light microscopy and found to be ～6.3 μm. P _d was measured by using an Mn²⁺‐doping 1H NMR (nuclear magnetic resonance) technique. The P _d (cm/s) values were relatively low: ～2.1×10^?3 at 25°C, 2.5×10^?3 at 30°C, 3.4×10^?3 at 37°C and 4.5 at 42°C for the platypus RBCs and ～2.8×10^?3 at 25°C, 3.2×10^?3 at 30°C, 4.5×10^?3 at 37°C and 5.7×10^?3 at 42°C for the crocodile RBCs. In parallel with the low water permeability, the E _a,d (activation energy of water diffusion) was relatively high, ～35 kJ/mol. These results suggest that “conventional” WCPs (water channel proteins), or AQPs (aquaporins), are probably absent from the plasma membranes of RBCs from both the platypus and the saltwater crocodile.     

Effect of Ca2+ on programmed death of guard and epidermal cells of pea leaves

The effect of Ca²⁺ on programmed death of guard cells (GC) and epidermal cells (EC) determined from destruction of the cell nucleus was investigated in epidermis of pea leaves. Ca²⁺ at concentrations of 1–100 μM increased and at a concentration of 1 mM prevented the CN—induced destruction of the nucleus in GC, disrupting the permeability barrier of GC plasma membrane for propidium iodide (PI). Ca²⁺ at concentrations of 0.1–1 mM enhanced drastically the number of EC nuclei stained by PI in epidermis treated with chitosan, an inducer of programmed cell death. The internucleosomal DNA fragmentation caused by CN^? was suppressed by 2 mM Ca²⁺ on 6 h incubation, but fragmentation was stimulated on more prolonged treatment (16 h). Presumably, the disruption of the permeability barrier of plasma membrane for PI is not a sign of necrosis in plant cells. Quinacrine and diphenylene iodonium at 50 μM concentration prevented GC death induced by CN^? or CN^? + 0.1 mM Ca²⁺ but had no influence on respiration and photosynthetic O₂ evolution in pea leaf slices. The generation of reactive oxygen species determined from 2′,7′-dichlorofluorescein fluorescence was promoted by Ca²⁺ in epidermal peels from pea leaves.     

The Effect of Leaf Water Potential, Leaf Temperature and Light Intensity on Leaf Diffusion Resistance and the Transpiration of Leaves of Malus sylvestris

The relationship between leaf resistance to water vapour diffusion and each of the factors leaf water potential, light intensity and leaf temperature was determined for leaves on seedling apple trees (Malus sylvestris Mill. cv. Granny Smith) in the laboratory. Leaf cuticular resistance was also determined and transpiration was measured on attached leaves for a range of conditions. Leaf resistance was shown to be independent of water potential until potential fell below — 19 bars after which leaf resistance increased rapidly. Exposure of leaves to CO₂-free air extended the range for which resistance was independent of water potential to — 30 bars. The light requirement for minimum leaf resistance was 10 to 20 W m^?2 and at light intensities exceeding these, leaf resistance was unaffected by light intensity. Optimum leaf temperature for minimum diffusion resistance was 23 ± 2°C. The rate of change measured in leaf resistance in leaves given a sudden change in leaf temperature increased as the magnitude of the temperature change increased. For a sudden change of 1°C in leaf temperature, diffusion resistance changed at a rate of 0.01 s cm^?1 min^?1 whilst for a 9°C leaf temperature change, diffusion resistance changed at a rate of 0.1 s cm^?1 min^?1. Cuticular resistance of these leaves was 125 s cm^?1 which is very high compared with resistances for open stomata of 1.5 to 4 s cm^?1 and 30 to 35 s cm^?1 for stomata closed in the dark. Transpiration was measured in attached apple leaves enclosed in a leaf chamber and exposed to a range of conditions of leaf temperature and ambient water vapour density. Peak transpiration of approximately 5 × 10^?6 g cm^?2 s^?1 occurred at a vapour density gradient from the leaf to the air of 12 to 14 g m^?3 after which transpiration declined due presumably to increased stomatal resistance. Leaves in CO₂-free air attained a peak transpiration of 11 × 10^?6 g cm^?2 s^?1 due to lower values of leaf resistance in CO₂ free air. Transpiration then declined in these leaves due to development of an internal leaf resistance (of up to 2 s cm^?1). The internal resistance was masked in leaves at normal CO₂ concentrations by the increase in stomatal resistance.     

Permeability evaluation of 45S5 Bioglass®-based scaffolds for bone tissue engineering

Permeability is a key parameter for microstructural design of scaffolds, since it is related to their capability for waste removal and nutrients/oxygen supply. In this framework, Darcy's experiments were carried out in order to determine the relationship between the pressure drop gradient and the fluid flow velocity in Bioglass^®-based scaffolds to obtain the scaffold's permeability. Using deionised water as working fluid, the measured average permeability value on scaffolds of 90–95% porosity was 1.96×10^?9 m². This value lies in the published range of permeability values for trabecular bone.     

Ecophysiology of cuticular transpiration: comparative investigation of cuticular water permeability of plant species from different habitats

Water permeabilities of astomatous, isolated cuticular membranes (CM) of 24 different plants species were measured. Permeances varied from 1.7×10^–11 m·s^–1 (Vanilla planifolia leaf) up to 2.1×10^–9 m·s^–1 (Malus cf. domestica fruit) among different plant species, thus covering a range of over 2 orders of magnitude. Ranking of species according to permeances resulted in four distinct groups. The first group, of species with the lowest cuticular transpiration rates, included evergreen species growing in warm dry tropical climates (e.g. Vanilla planifolia and Monstera deliciosa leaves). The second class, with slightly higher water permeabilities, included evergreen species with typical scleromorphic leaf properties, adapted to a typical mediterranean type of climate with a dry period during the year (e.g. Citrus limon and Olea europaea leaves). The third group of species, where the highest leaf cuticular transpiration rates were observed, included deciduous species normally growing in a tempeate climate (e.g. Juglans regia and Forsythia suspensa leaves). Fruit cuticular membranes (CM) made up the fourth group (e.g. Capsicum annuum and Malus cf. domestica fruits), with even higher permeances than leaves of species from group 3. Thus, it appears that the plant species investigated show ecophysiological adaptations to the climatic demands of their natural habitats in cuticular water permeability.     

Effects of diet, body size, age and temperature on growth rates in the amphipod Gammarus pulex

SUMMARY. Increase in body wet weight of Gammarus pulex fed on decaying elm leaves was followed to senescence and death. Growth in juveniles was approximately exponential; from birth to death it conformed to a logistic growth curve, with maximum absolute increments in weight about half-way through a life span of 350–450 days at 15°C. Some individuals lived longer, for up to 640–700 days. The instantaneous or specific growth rate was maximal near birth, at c. 5–6% wet wt day^?1, and declined exponentially with increasing size and age. Over the range 4.7–14.8°C there was a log-log relationship between temperature and specific growth rate. Growth was maximal at 20°C in newborn animals and at 15°C in 6–9-mg animals. The specific growth rate of young individuals was fastest on decaying leaves of elm with a well developed flora of fungi and other microorganisms. Leached elm leaves without this flora supported growth at a lower rate. The latter diet was sufficient for survival and growth of newborn individuals; detritus, faeces or other food items were not needed. Isolated specimens grew as fast as those kept in groups. Growth was generally slower on leached leaves of oak and sycamore. In newborn animals fed on the fine roots of aquatic plants (Veronica, Rorippa and Glyceria), growth was as fast as on decaying elm leaves; growth on the green living leaves of the plants was slower, as on detritus from two streams and on a pure culture of an aquatic fungus. Consumption of leached elm leaves was related to leaf thickness. In a full gut the wet weight (1.34–1.37 mg) and volume (3.8–4.1 mm³) (for 20-mg animals) was independent of leaf thickness but dependent on animal size, increasing 4-fold over the range 2–50 mg body wt. Daily consumption (dry wt) was approximately equivalent to 50% body dry wt at 5 mg and 20% at 50 mg body wet wt. Individuals fed on thick leaves ingested 50% more dry weight per day and absorbed more in the gut than when fed on thin leaves, but the relative efficiency of absorption was the same at 36–59% for 10–20-mg animals. Weight-specific absorption in the gut was highest in juveniles and decreased with increasing body weight; relative efficiency of absorption was generally lower in the larger individuals. Assuming an energy value of 5 cal mg^?1 dry wt for elm leaves, daily mean energy intake by absorption in thegutof G. pu/ex was2.2 cal mg^?1 animaldry wt (9.2 J mg^?1) in individuals of 0.4 mgdry wt (2 mg wet wt), decreasing to 0.3 cal mg^?1 (1.3 J mg^?1) at 10 mg dry wt (50 mg wet wt). Growth in Gammarus is briefly reviewed in the hght of work on other animals and it is emphasized that all aspects of feeding, growth and metabol-ism should be specifically related to size and age of the individuals, using well defined diets.     

Wnt11 preserves mitochondrial membrane potential and protects cardiomyocytes against hypoxia through paracrine signaling

We investigated the effect of Wnt11 on mitochondrial membrane integrity in cardiomyocytes (CMs) and the underlying mechanism of Wnt11-mediated CM protection against hypoxic injury. A rat mesenchymal stem cell (MSC) line that overexpresses Wnt11 (MSC^Wnt11) and a control cell line transduced with empty vector (MSC^Null) were established to determine the cardioprotective role of Wnt11 in response to hypoxia. Mitochondrial membrane integrity in MSC^Wnt11 cells was assessed using fluorescence assays. The role of paracrine signaling mediated by vascular endothelial growth factor (VEGF), basic fibroblast growth factor (b-FGF), and insulin-like growth factor 1 (IGF-1) in protecting CMs against hypoxia were investigated using cocultures of primary CMs from neonatal rats with conditioned medium (CdM) from MSC^Wnt11. MSC^Wnt11 cells exposed to hypoxia reduced lactate dehydrogenase release from CMs and increased CM survival under hypoxia. In addition, CMs cocultured with CdM that were exposed to hypoxia showed reduced CM apoptosis and necrosis. There was significantly higher VEGF and IGF-1 release in the MSC^Wnt11 group compared with the MSC^Null group, and the addition of anti-VEGF and anti-IGF-1 antibodies inhibited secretion. Moreover, mitochondrial membrane integrity was maintained in the MSC^Wnt11 cell line. In conclusion, overexpression of Wnt11 in MSCs promotes IGF-1 and VEGF release, thereby protecting CMs against hypoxia.     

Interaction of Growth Retardants and Temperature in Growth, Flowering, Regeneration, and Auxin Activity of Begonia × cheimantha Everett

Soil application of CCC reduced stem and leaf growth in Begonia plants. This effect was evident with all concentrations tested at 18°C, whereas at 21 and 24°C no growth–retarding effect was observed with 2 × 10^?2 M CCC, and with 5 × 10^?3 M growth was even stimulated. Flowering was promoted by CCC in long day and neur–critical temperature, particularly under low light intensity in the winter. The formation of adventitious buds in leaves of plants grown at 21 and 24°C was stimulated when the plants received 5 × 10^?2 and 2 × 10^?2 M CCC, while 8 7times; 10^?2 M was inhibitory. In plants grown at 18°C bud formation was inhibited by all CCC concentrations. Root formation in the the leaves was usually stimulated by high CCC concentrations, while root elongation was reduced. The level of ether–extractable. acidic auxin (presumably IAA) in the leaves was lowered by CCC treatment of the plants, hut this required higher CCC concentrations at higt than at low temperature. When applied to detached leaves CCC stimulated bud formation at concentrations ranging from 10^?4 to 10^?2 M in leaves planted at 18 and 21°C. At 24°C budding was inhibited by 10^?2 M CCC, the lower concentrations being stimulatory also at this temperature. Root formation and growth were not much affected by CCC treatment of the leaves, but increased with the temperature. Soil application of Phosfon (4 × 10^?4 M) had no effect on growth and flowering, nov did it affect the subsequent regeneration of buds and roots in the leaves. In detached leaves Phosfon stimulated bud formation with au optimum at 10^?6 M. Root formation was stimulated by Phosfon at all temperatures, the optimal concentration being 10^?5 M, whereas root length was conversely affected. Foliar application of B-995 to intact plants and treatment of detached leaves greatly inhibited the formation of buds and had little effect on root formation. B-99D reduced the growth and delayed flowering in the plants.     

pH-Induced Proton Permeability Changes of Plasma Membrane Vesicles

In vivo studies with leaf cells of aquatic plant species such as Elodea nuttallii revealed the proton permeability and conductance of the plasma membrane to be strongly pH dependent. The question was posed if similar pH dependent permeability changes also occur in isolated plasma membrane vesicles. Here we report the use of acridine orange to quantify passive proton fluxes. Right-side out vesicles were exposed to pH jumps. From the decay of the applied ΔpH the proton fluxes and proton permeability coefficients (P_H+) were calculated. As in the intact Elodea plasma membrane, the proton permeability of the vesicle membrane is pH sensitive, an effect of internal pH as well as external pH on P_H+ was observed. Under near symmetric conditions, i.e., zero electrical potential and zero ΔpH, P_H+ increased from 65 × 10⁻⁸ at pH 8.5 to 10⁻¹ m/sec at pH 11 and the conductance from 13 × 10⁻⁶ to 30 × 10⁻⁴ S/m². At a constant pH _i of 8 and a pH _o going from 8.5 to 11, P_H+ increased more than tenfold from 2 to 26 × 10⁻⁶ m/sec. The calculated values of P_H+ were several orders of magnitude lower than those obtained from studies on intact leaves. Apparently, in plasma membrane purified vesicles the transport system responsible for the observed high proton permeability in vivo is either (partly) inactive or lost during the procedure of vesicle preparation. The residue proton permeability is in agreement with values found for liposome or planar lipid bilayer membranes, suggesting that it reflects an intrinsic permeability of the phospholipid bilayer to protons. Possible implications of these findings for transport studies on similar vesicle systems are discussed. Received: 5 April 1995/Revised: 28 March 1996     

6‐Methyl 3‐chromonyl 2,4‐thiazolidinedione/2,4‐imidazolidinedione/2‐thioxo‐imidazolidine‐4‐one compounds: novel scavengers of reactive oxygen species

The benefits of antioxidants on human health are usually ascribed to their potential ability to remove reactive oxygen species providing protection against oxidative stress. In this paper the free radicals scavenging activities of nine 6‐methyl 3‐chromonyl derivatives (CMs) were evaluated for the first time by the chemiluminescence, electron paramagnetic resonance, spin trapping and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH^?) methods. The total antioxidant capacity was also measured using a ferric‐ferrozine reagent. Compounds having a hydrogen atom at the N3‐position of the β‐ring were effective in quenching CL resulted from the KO₂/18‐crown‐6‐ether system (a source of superoxide anion radical, ) in a dose‐dependent manner over the range of 0.05–1 mmol/L [IC₅₀ ranged from 0.353 (0.04) to 0.668 (0.05) mmol/L]. The examined compounds exhibited a significant scavenging effect towards hydroxyl radicals (HO^? HO^?), produced by the Fenton reaction, and this ranged from 24.0% to 61.0%, at the concentration of 2.5 mmol/L. Furthermore, the compounds examined were also found to inhibit DPPH^? and this ranged from 51.9% to 97.4% at the same concentration. In addition, the use of the total antioxidant capacity assay confirmed that CM compounds are able to act as reductants. According to the present study, CM compounds showed effective in vitro free radical scavenging activity and may be considered as potential therapeutics to control diseases of oxidative stress‐related etiology. Copyright © 2013 John Wiley & Sons, Ltd.     

Comparative NMR studies of diffusional water permeability of red blood cells from different species: XVI Dingo (Canis familiaris dingo) and dog (Canis familiaris)

As part of a programme of comparative measurements of P _d (diffusional water permeability) the RBCs (red blood cells) from dingo (Canis familiaris dingo) and greyhound dog (Canis familiaris) were studied. The morphologies of the dingo and greyhound RBCs [examined by light and SEM (scanning electron microscopy)] were found to be very similar, with regard to aspect ratio and size; the mean diameters were estimated to be the same (～7.2 μm) for both dingo and greyhound RBCs. The water diffusional permeability was monitored by using an Mn²⁺‐doping ¹H NMR technique at 400 MHz. The P _d (cm/s) values of dingo and greyhound RBCs were similar: 6.5×10^?3 at 25°C, 7.5×10^?3 at 30°C, 10×10^?3 at 37°C and 11.5×10^?3 at 42°C. The inhibitory effect of a mercury‐containing SH (sulfhydryl)‐modifying reagent PCMBS (p‐chloromercuribenzene sulfonate) was investigated. The maximal inhibition of dingo and greyhound RBCs was reached in 15–30 min at 37°C with 2 mmol/l PCMBS. The values of maximal inhibition were in the range 72–74% when measured at 25°C and 30°C, and ～66% at 37°C. The lowest value of P _d (corresponding to the basal permeability to water) was ～2–3×10^?3 cm/s in the temperature range 25–37°C. The E _a,d (activation energy of water diffusion) was 25 kJ/mol for dingo RBC and 23 kJ/mol for greyhound RBCs. After incubation with PCMBS, the values of E _a,d increased, reaching 46–48 kJ/mol in the condition of maximal inhibition of water exchange. The electrophoretograms of membrane polypeptides of the dingo and greyhound RBCs were compared and seen to be very similar. We postulate that the RBC parameters reported in the present study are characteristic of all canine species and, in particular in the two cases presented here, these parameters have not been changed by the peculiar Australian habitat over the millennia (as in the case of the dingo) or over shorter time periods, decades or centuries (as in the case of the domestic greyhound).     

Salt resistance in two cashew species is associated with accumulation of organic and inorganic solutes

This study establishes relationships between salt resistance and solute accumulation in roots and leaves of two contrasting cashew species. The sensitive (Anacardium microcarpum) and resistant (A. occidentale) species showed maximum root LD₅₀ values (the external NaCl concentration required for a 50% reduction in dry weight) of 63 and 128?mM NaCl, whereas the shoot LD₅₀ values were 90 and 132?mM, respectively. The salt sensitivity was directly associated with Na⁺ accumulation and especially with the Cl^? content in leaves and to a minor extent in roots. The accumulation of saline ions was associated with higher net uptake rates by roots and transport rates from root to shoot in the sensitive cashew species. The K⁺/Na⁺ ratios were not associated with salt resistance either in roots or leaves. Proline and free amino acid concentrations were strongly increased by salinity, especially in the leaves of the resistant species. The soluble sugar concentrations were not influenced by NaCl treatments in leaves of both species. In contrast, the root soluble sugar content was significantly decreased by salinity in the sensitive species only. In conclusion, the higher salt sensitivity of A. microcarpum is associated to an inefficient salt exclusion system of the leaves, especially for Cl^?. On the other hand, the resistant species displays higher concentrations of organic solutes especially a salt-induced accumulation of proline and free amino acids in leaves.     

Diversity and activity of aquatic fungi under low oxygen conditions

1. The objective was to test whether a decrease in oxygen concentration in streams affects the diversity and activity of aquatic hyphomycetes and consequently leaf litter decomposition. 2. Senescent leaves of Alnus glutinosa were immersed for 7 days in a reference stream, for fungal colonization, and then incubated for 18 days in microcosms at five oxygen concentrations (4%, 26%, 54%, 76% and 94% saturation). Leaf decomposition (as loss of leaf toughness), fungal diversity, reproduction (as spore production) and biomass (ergosterol content) were determined. 3. Leaf toughness decreased by 70% in leaves exposed to the highest O₂ concentration, whereas the decrease was substantially less (from 25% to 45%) in treatments with lower O₂. Fungal biomass decreased from 99 to 12 mg fungi g⁻¹ ash‐free dry mass on exposure to 94% and 4% O₂ respectively. Sporulation was strongly inhibited by reduction of dissolved O₂ in water (3.1 × 10⁴ versus 1.3 × 10³ spores per microcosms) for 94% and 4% saturation respectively. 4. A total of 20 species of aquatic hyphomycetes were identified on leaves exposed to 94% O₂, whereas only 12 species were found in the treatment with 4% O₂ saturation. Multidimensional scaling revealed that fungal assemblages exposed to 4% O₂ were separated from all the others. Articulospora tetracladia, Cylindrocarpon sp. and Flagellospora curta were the dominant species in microcosms with 4% O₂, while Flagellospora curvula and Anguillospora filiformis were dominant at higher O₂ concentrations. 5. Overall results suggest that the functional role of aquatic hyphomycetes as decomposers of leaf litter is limited when the concentration of dissolved oxygen in streams is low.