Azathioprine-induced in vitro inhibition of prostaglandin E2 production by rabbit retina

Although much work exists concerning the clinical and immunosuppressive effects of azathioprine, little is known of the mechanism of action of this drug. The present study reports that azathioprine at the concentrations of 0.1 and 0.05 micrograms/ml causes a significant suppression of in vitro prostaglandin E2 production by rabbit retinas. However, at concentrations below (0.01 microgram/ml) or above (1,10 micrograms/ml) azathioprine was not inhibitory effect. These results suggest a dose-dependent inhibitory effect of azathioprine on prostaglandin E2 production.     

Identification of the Cl− transport site of human red blood cells by a kinetic analysis of the inhibitory effects of a chemical probe

H₂DIDS, the dihydro analog of DIDS (4,4′-diisothiocyanostilbene-2,2′-disulfonic acid) can interact covalently with membrane sites, resulting in an irreversible inhibition of anion exchange. At low temperatures (0°C) and for relatively short times, however, its interaction is largely reversible, so that a kinetic analysis of the nature of its inhibitory effect on Cl^? self exchange can be performed. The effects of variations in the chloride concentration on the inhibitory potency of H₂DIDS are consistent with the concept that Cl^? and H₂DIDS compete for the transport site of the anion exchange system. The value of K_i for H₂DIDS is 0.046 μM, indicating that H₂DIDS has a higher affinity for the transport system than any other inhibitor so far examined. If, as seems probable, the covalent labelling of H₂DIDS occurs at the same site as the reversible binding, H₂DIDS can be used as a covalent label for the transport site. The specific localization of H₂DIDS in the band-3 protein thus indicates that this protein participates directly in anion exchange.     

Effect of PGPR on population growth parameters of cotton aphid

Plant-growth-promoting rhizobacteria (PGPR) help plants by increasing plant growth and defending them against pathogens and pests. There is considerable research supporting the use of these bacteria in plant pathogens management, but the number of research papers that have focused on their effect on insect pests control is quite few. In this research, we examined the ability of four native strains of Pseudomonas fluorescens to manage the cotton aphid, Aphis gossypii, on greenhouse-grown cucumbers in Iran. The strains UTPF (University of Tehran Pseudomonas fluorescens) 68, UTPF1, UTPF6 and PF169 of P. fluorescens were used as seed treatment to evaluate their impact on both cucumber growth and cotton aphid population growth parameters. The strains UTPF68 and PF169 of this bacterium influenced phenology of cucumber by decreasing the Growing Degree Days of flowering period. Also, the plants treated with these bacterial strains had a significant increase in yield weight of approximately 58%. In the other part of the study, various effects on developmental parameters of aphid were recorded among bacterial strains, but only PF169 caused significant decreasing effect on r_m value of aphid, which is the most important factor in aphid’s life table. We conclude that the PF169 treatment had significant negative effects on population growth rate of A. gossypii. The PF169 could be reliable in a control programme for A. gossypii on cucumber plants in greenhouses.     

The thylakoid lumen protease Deg1 is involved in the repair of photosystem II from photoinhibition in Arabidopsis

Deg1 is a Ser protease peripherally attached to the lumenal side of the thylakoid membrane. Its physiological function is unknown, but its localization makes it a suitable candidate for participation in photoinhibition repair by degradation of the photosystem II reaction center protein D1. We transformed Arabidopsis thaliana with an RNA interference construct and obtained plants with reduced levels of Deg1. These plants were smaller than wild-type plants, flowered earlier, were more sensitive to photoinhibition, and accumulated more of the D1 protein, probably in an inactive form. Two C-terminal degradation products of the D1 protein, of 16 and 5.2 kD, accumulated at lower levels compared with the wild type. Moreover, addition of recombinant Deg1 to inside-out thylakoid membranes isolated from the mutant could induce the formation of the 5.2-kD D1 C-terminal fragment, whereas the unrelated proteases trypsin and thermolysin could not. Immunoblot analysis revealed that mutants containing less Deg1 also contain less FtsH protease, and FtsH mutants contain less Deg1. These results suggest that Deg1 cooperates with the stroma-exposed proteases FtsH and Deg2 in degrading D1 protein during repair from photoinhibition by cleaving lumen-exposed regions of the protein. In addition, they suggest that accumulation of Deg1 and FtsH proteases may be coordinated.     

An N-terminal nucleotide-binding site in VDAC1: involvement in regulating mitochondrial function

In a previous study, we presented evidence for the existence of a nucleotide-binding site (NBS) in the N-terminal region of the voltage-dependent anion channel (VDAC1). In this study, further localization and possible roles of the proposed VDAC1-NBS were investigated using site-directed mutagenesis. The predicated NBS of murine VDAC1 (mVDAC1) was mutated by replacing two glycine residues with alanines or a conserved lysine residue with a serine. Expression of the G21A,G23A- and K20S-mVDAC1s in human T-REx-293 cells in which endogenous VDAC1 expression had been silenced affected cell growth and cytosolic ATP levels. While G21A,G23A-mVDAC1-expressing cells displayed growth rates similar to native-mVDAC1-expressing cells, the K20S-mVDAC1-expressing cells displayed significantly retarded growth and increased resistance to cell death. Cells expressing either mVDAC1 mutant also displayed significantly reduced cellular ATP levels. When K20S-mutant mVDAC1 was expressed in porin1-less yeast, the transformed cells grew slower on non-fermentable carbon sources, while isolated mitochondria expressing either mVDAC1 mutant showed significant reduction in ATP synthesis. Purified K20S-mVDAC1 displayed a significant decrease in [alpha-(32)P]BzATP-binding and altered channel properties, that is, reduced ion selectivity, while the G21A,G23A-mutant protein displayed only a mild reduction in channel selectivity. These results suggest that mutations in the proposed VDAC1-NBS, particularly the K20S, altered channel activity, thereby interfering with VDAC function as the major pathway for the transport of metabolites and adenine nucleotides across the outer mitochondrial membrane. Finally, involvement of the VDAC1-NBS in the control of mitochondrial ATP synthesis, cell growth and viability is discussed.     

Developing potent backbone cyclic peptides bearing the shared epitope sequence as rheumatoid arthritis drug-leads

Rheumatoid arthritis (RA) is a common human leukocyte antigen-associated disease. Most RA patients have a five-residue sequence motif called the shared epitope (SE) in the DRβ-chain of the HLA-DRB1 protein. The SE was found to activate nitric oxide (NO) production, suggesting a possible mechanism for RA development. The native conformation of the SE is presumed to be an α-helix, thus using cyclic peptides to stabilize this conformation may produce a potent SE mimetic which will have drug-like properties. We present the development of a backbone cyclic SE mimetic that activates NO production in the low nM range. Circular dichroism analysis revealed a conformational change from for the parent linear peptides to the cyclic analogs. The most active cyclic analog is completely stable towards trypsin/chymotrypsin degradation while the linear 15-mer analogs completely degraded within 30 min. The outcome of this study is a potent cyclic peptide with drug-like properties that can be used as a template for drug development.     

Quantal release of glutamate generates pure kainate and mixed AMPA/kainate EPSCs in hippocampal neurons

The relative contribution of kainate receptors to ongoing glutamatergic activity is at present unknown. We report the presence of spontaneous, miniature, and minimal stimulation-evoked excitatory postsynaptic currents (EPSCs) that are mediated solely by kainate receptors (EPSC(kainate)) or by both AMPA and kainate receptors (EPSC(AMPA/kainate)). EPSC(kainate) and EPSC(AMPA/kainate) are selectively enriched in CA1 interneurons and mossy fibers synapses of CA3 pyramidal neurons, respectively. In CA1 interneurons, the decay time constant of EPSC(kainate) (circa 10 ms) is comparable to values obtained in heterologous expression systems. In both hippocampal neurons, the quantal release of glutamate generates kainate receptor-mediated EPSCs that provide as much as half of the total glutamatergic current. Kainate receptors are, therefore, key players of the ongoing glutamatergic transmission in the hippocampus.     

The effect of fire on nutrients in a pine forest soil

The effect of a hot summer fire on soil nutrient contents in the upper 2 cm of Aleppo pine forest with a dense woody understory was studied from September 1985 to May 1986. In comparison with the adjacent unburned forest, total nitrogen decreased by 25% but available forms of nitrogen were much higher. In burned and unburned soils there was a similar trend to increase and decrease in NH ₄ ⁺ −N, However, while (NO ₂ ⁻ +NO ₃ ⁻ −N decreased in the unburned soil it rose rapidly in the burned ash soil. Total phosphorus increased by 300% after the fire but decreased again 2 months later. Also water-soluble P increased up to November and then decreased to the levels of the unburned soils. The same was true for electrical conductivity and pH, increasing immediately after the fire and then leveling off again. This increase in nutrient levels in the “ash soil” was reflected in the striking increase in shoot and root biomass and in the content of N, P, Mg, K, Ca, Zn and Fe in wheat and clover plants grown in pots in these soils. These nutrient levels were much higher in the wheat plants, which also produced 12 times more seeds in the “ash soil.” It seems that fire in these pine forests causes a short-term flush of the mineral elements in the upper “ash soil” layer which is reverted gradually via the herbaceous post-fire to the ecosystem.     

Ecological and Cultural Landscape Restoration and the Cultural Evolution towards a Post-Industrial Symbiosis between Human Society and Nature

I discuss ecological and cultural restoration within the broader context of the critical transition period from the fossil fuel age to the post-industrial global information age. In this cultural evolutionary process, the restoration of natural and cultural landscapes should play a vital role. For this purpose, it has to be guided by a holistic and transdisciplinary systems approach, aiming not only at the organismic but also at the functional and structural restoration of ecological and cultural diversity as total landscape ecodiversity. For the development of suitable restoration strategies, a clear distinction has to be made between different functional classes of natural and cultural solar-powered biosphere and fossil-powered technosphere landscapes, according to their inputs and throughputs of energy and materials, their organisms, their control by natural or human information, their internal self-organization and their regenerative capacities. Not only technosphere landscapes but also intensive agro-industrial landscapes have lost these capacities and are heavily subsidized by fossil energy and chemicals, to the detriment of the environment and human health. They therefore have to be rehabilitated by more sustainable but not less productive agricultural systems based on organic farming. But their natural regenerative capacities can be restored only by regenerative systems, with the help of cultural "neotechnic" information. The promise for an urgently required evolutionary symbiosis between human society and nature in a sustainable post-industrial total human ecosystem lies in the functional integration of such innovative regenerative systems and all natural and cultural biosphere landscapes with healthier and more livable technosphere landscapes. To this goal, ecological and cultural landscape restoration can make an important contribution.