Effects of Ca, Cu, Al and La on pectin gel strength: implications for plant cell walls

Rheology of Ca-pectate gels is widely studied, but the behaviour of pectate gels formed by Cu, Al and La is largely unknown. It is well known that gel strength increases with increasing Ca concentration, and it is hypothesised that this would also be the case for other cations. Pectins are a critical component of plant cell walls, imparting various physicochemical properties. Furthermore, the mechanism of metal toxicity in plants is hypothesised to be, in the short term, related to metal interactions with cell wall pectin. This study investigated the influence of Ca, Cu, Al and La ion concentrations at pH 4 on the storage modulus as a function of frequency for metal-pectin gels prepared from pectin (1%) with a degree of esterification of 30%. Gels were formed in situ over 6 d in metal chloride solution adjusted daily to pH 4. Cation concentration was varied to develop a relationship between gel strength and cation concentration. At similar levels of cation saturation, gel strength increased in the order of La < Ca ? Al ? Cu. The swelling of the gels also varied between cations with Ca gels being the most swollen.     

Urgency,development stage and coordination degree analysis to support differentiation management of water pollution emission control and economic development in the eastern coastal area of China

The eastern coastal areas of China have high-density population, developed society and economy, and large water pollution emissions. How to reduce water pollution and realize the coordinated development of the economy and environment has become the national focus. Effective environmental policies should consider regional differences in development stage and sustainability performance. Here, we first analyzed the water pollution emissions intensity of the eastern coastal areas of China and the urgency of emissions reduction using 8-year environmental statistics from 2003 to 2010. We characterized development stages of the eastern coastal areas based on the relationships between water pollution emissions intensity and economic development. Further, we built a coordination degree index of economic development and water environment protection as a measure of sustainability. Results show that water pollution emissions intensity decreases as the economy grows from 2003 to 2010. The less-developed regions have a better coordination degree than some more-developed regions, especially those most-developed ones (e.g., Shanghai show more pressures on long-term sustainability than Hebei). The less-developed regions should take advantage of economic growth to invest more advanced environment protection technologies. The more-developed regions need to upgrade its economic structures and municipal infrastructures. Overall, the study provided a comprehensive approach to understand regional difference in development stage and sustainability performance in the eastern coastal region of China as well as the need of different environmental policies to reduce water pollution emissions.     

Partial complex I inhibition decreases mitochondrial motility and increases matrix protein diffusion as revealed by fluorescence correlation spectroscopy

We previously reported that inhibition of mitochondrial complex I (CI) by rotenone induces marked increases in mitochondrial length and degree of branching, thus revealing a relationship between mitochondrial function and shape. We here describe the first time use of fluorescence correlation spectroscopy (FCS) to simultaneously probe mitochondrial mobility and intra-matrix protein diffusion, with the aim to investigate the effects of chronic CI inhibition on the latter two parameters. To this end, EYFP was expressed in the mitochondrial matrix of human skin fibroblasts (mitoEYFP) using baculoviral transduction and its diffusion monitored by FCS. This approach revealed the coexistence of moving and stationary mitochondria within the same cell and enabled simultaneous quantification of mitochondrial velocity and mitoEYFP diffusion. When CI activity was chronically reduced by 80% using rotenone treatment, the percentage of moving mitochondria and their velocity decreased by 30%. MitoEYFP diffusion did not differ between moving and stationary mitochondria but was increased 2-fold in both groups of mitochondria following rotenone treatment. We propose that the increase in matrix protein diffusion together with the increase in mitochondrial length and degree of branching constitutes part of an adaptive response which serves to compensate for the reduction in CI activity and mitochondrial motility.     

Activation of Murine Macrophages by Polysaccharide Fractions from Marine Algae (Porphyra yezoensis)

Two discrete immunomodulating fractions were obtained from marine algae (Porphyra yezoensis): one was the Porphyra water-soluble fraction (PWSF) which was extracted with hot water from the whole body of algae, and the other was the Porphyra acid-soluble fraction (PASF) which was extracted with acid from the residue. The major constituent in both PWSF and PASF was a polysaccharide, the total sugar concentration in PWSF (56.4%) being lower than that in PASF (82.2%). The high contents of 3,6-anhydrogalactose and sulfate indicated the porphyran structure in PWSF and PASF. The results of an in vitro culture assay with proteose peptone-induced macrophages from mice revealed that PWSF and PASF both enhanced glucose consumption, as well as the production of nitrite and tumor necrosis factor (TNF), but that these were increased more by PWSF than by PASF. PWSF augmented IL-1 secretion from these macrophages, while PASF did not. On the other hand, the carbon clearance activity of phagocytes from mice injected intraperitoneally with PASF was higher than that from PWSF-injected mice. The injection of PASF into mice also enhanced the carbon clearance activity in a dose-dependent manner. These results suggest that the two individual fractions possessed the ability to activate macrophages in vitro and in vivo in different ways.     

Emission-wavelength-dependent decay of the fluorescent probe N-phenyl-1-naphthylamine

We have measured the fluorescence decay of $\text{N-}\text{phenyl}\text{-1-}\text{naphthylamine}$ using the phase-modulation method, in several solvent systems and egg phosphatidylcholine vesicles. The decay is monoexponential in pure solvents (both polar and non-polar) of low viscosity. In polar viscous solvents or in non-polar solvents containing an added polar solute, the decay is heterogeneous and emission wavelength dependent. In such cases, dielectric relaxation and/or excited-state complexing give rise to a shift of the emission spectrum on the nanosecond time scale. Emission-wavelength-dependent decay was also observed when $\text{N-}\text{phenyl}\text{-1-}\text{naphthylamine}$ was bound to egg phosphatidylcholine vesicles. From these results as well as the position of the emission spectral maximum, we conclude that $\text{N-}\text{phenyl}\text{-1-}\text{naphthylamine}$ probes the ester-carbonyl region of the phospholipid acyl chains, where it undergoes an excited-state reaction. This result contradicts the often made assumption that $\text{N-}\text{phenyl}\text{-1-}\text{naphthylamine}$ probes the deeper hydrocarbon region of the bilayer.     

Measurement of fluxes through the pentose phosphate pathway in erythrocytes from individuals with sickle cell anemia by carbon-13 nuclear magnetic resonance spectroscopy

Erythrocytes from individuals with sickle cell anemia have previously been shown to have increased levels of intracellular oxidants and increased oxidative damage. Oxidative damage has been implicated in the events leading to the painful crises and hemolytic anemia found in sickle cell anemia. Since the pentose phosphate pathway (PPP) is an important source of reducing capacity in erythrocytes, we have investigated the fluxes through the PPP in normal and sickle cell erythrocytes using [2-¹³C]D-glucose and carbon-13 nuclear magnetic resonance (NMR) spectroscopy. Our results indicate that sickle cell erythrocytes have a flux through the PPP of 0.13±0.02 μmol/h per ml erythrocytes that is comparable to that in normal erythrocytes, 0.21±0.02 μmol/h per ml erythrocytes. However, when stimulated with methylene blue, sickle cell erythrocytes show a decreased response, 0.59±0.10 μmol/h per ml erythrocytes, compared to normal erythrocytes, 1.64±0.10 μmol/h per ml erythrocytes. When homogeneous populations of sickle cell erythrocytes are isolated by density gradient centrifugation, the rate of flux through the PPP in methylene blue-stimulated sickle cell erythrocytes, 1.16±0.16 μmol/h per ml erythrocytes, approaches that in methylene blue-stimulated normal erythrocytes. In addition, by analyzing the dose response to methylene blue, we have found that the decreased stimulation of the PPP by methylene blue in heterogeneous populations of sickle cell erythrocytes is a failure of methylene blue to simulate the PPP rather than a deficiency in the PPP in sickle cell erythrocytes.     

Thermal stability of a flavoprotein assessed from associative analysis of polarized time-resolved fluorescence spectroscopy

Upon gradually heating a particular mutant of the flavoprotein NADH peroxidase, it was found from the peculiar time-resolved fluorescence anisotropy pattern of the flavin prosthetic group (FAD) that, at elevated temperature, FAD is released from the tetrameric enzyme. Since in this case a mixture of free and enzyme-bound FAD contributes to the time-dependent fluorescence anisotropy, its analysis can only be accomplished by an associative fitting model, in which specific fluorescence lifetimes of both species are linked to specific correlation times. In this letter the general approach to the associative polarized fluorescence decay analysis is described. The procedure can be used for other flavoproteins to determine the temperature at which the onset of thermal denaturation will start, leading to release of the flavin prosthetic group. Received: 20 November 1998 / Revised version: 6 April 1999 / Accepted: 8 April 1999