Electron donation in Photosystem II

The electron transfer resulting from illumination and dark storage of PS II has been studied using EPR signals from several electron carriers. The recombination of D⁺ (Signal II) and Q⁻_A formed by illumination occurred during dark storage at 77 K and was used to deplete reaction centres of D⁺. The donor D was then shown to be oxidized in the dark by the S₂ state of the oxygen-evolving complex. A slow change which occurred during dark storage of PS II samples was detected using the power saturation characteristics of D. We interpret this effect on D to be an indirect result of a rearrangement of the manganese complex during long-term dark adaptation. A role for D in the stability, protection and perhaps initial manganese binding of the oxygen-evolving complex is suggested.     

Anaerobic degradation of papermill sludge in a two-phase digester containing rumen microorganisms and colonized polyurethane foam

Summary The use of reticulated polyurethane foam as a support material for the immobilization of methanogenic associations and its application to the anaerobic treatment of fine particulate solid wastes was investigated. The colonization of polyurethane support particles in a continuous upflow reactor fed on a mixture of acetate, propionate and butyrate, was both rapid and dense. The combination of rumen microorganisms and colonized support particles in a two-phase digester resulted in an efficient anaerobic decomposition of papermill sludge.     

A genetically related response to iron deficiency stress in muskmelon

A mutant muskmelon (Cucumis melo L.) with characteristic Fe-deficiency chlorosis symptoms was compared to related cultivars in its ability to obtain Fe via the widely known Fe-stress response mechanisms of dicotyledonous plants. The three cultivars (fefe, the Fe-inefficient mutant; Mainstream and Edisto, both Fe efficient plants) were grown in nutrient solution in either 0 or 3.5 mg L^-1 Fe as FeCl₃. None of the three cultivars released reductants or phytosiderophores, but both Edisto and Mainstream produced massive amounts of H+ ions to reduce and maintain the pH of nutrient solutions below pH 4.0. The roots of these two Fe-efficient cultivars were also capable of reducing Fe³⁺ to Fe²⁺. These responses maintained green plants, resulted in high leaf Fe in both Edisto and Mainstream, and produced Mn toxicity in Mainstream. The lack of Fe-deficiency stress response in fefe not only affected leaf Fe concentration and chlorosis, but also resulted in reduced uptake of Mn. The importance of reduced Fe (Fe²⁺) to the Fe-efficient cultivars was confirmed by growing the cultivars with BPDS (4, 7-diphenyl-1, 10-phenanthroline disulfonic acid, a ferrous chelator) and EDDHA [ethylene-diamine di (0-hydroxphenylacetic acid)] (a ferric chelator), and observing increased chlorosis and reduced Fe uptake in BPDS grown plants. The Fe-deficiency response observed in these cultivars points out the diversity of responses to Fe deficiency stress in plants. The fefe mutant has a limited ability to absorb Fe and Mn and perhaps could be used to better understand Mn uptake in plants.     

International scientific meetings: relation between structure and function.

Increasing pressure is being placed on the scientific community to evaluate research activities. Scientific meetings consume a small but important fraction of the research budget. Audit of a well established series of scientific meetings showed that they met their immediate objectives in that they were international and multidisciplinary and provided a forum in which all participants actively contributed to discussion. The meetings had a positive outcome for the participants, leading in many cases to the subsequent exchange of research material (60% of participants) and to the establishment of collaborative research projects (31%). The impact of the meetings on the scientific community at large was assessed by citation analysis, which showed that the proceedings were cited early, often, and over a substantial period.     

A radioimmunoassay for'plasma progesterone

A radioimmunoassay for progesterone was developed which uses one micro-column chromatography for purification of hexane extracts of plasma.     

Kinetics of basic fibroblast growth factor binding to its receptor and heparan sulfate proteoglycan: a mechanism for cooperactivity.

Basic fibroblast growth factor (bFGF) binds to cell surface receptor (CSR) proteins and to heparan sulfate proteoglycans (HSPG). On the basis of equilibrium dissociation constants (Kd), the CSR has been considered a "high-affinity" binding site and HSPG a "low-affinity" site. We measured the apparent individual on and off rate constants (kon and koff) for bFGF binding to these two sites on intact cells and to each class of binding site in the absence of the other. While the kon's for CSR and HSPG on intact cells were not statistically different (konC = 2.27 x 10(8) M-1 min-1; konH = 0.90 x 10(8) M-1 min-1), the koff for the HSPG was 22.7-fold greater than that for the CSR (koffC = 0.003 min-1; koffH = 0.68 min-1). Thus, the difference in Kd's appears to result from the faster rate at which bFGF is released from the HSPG sites compared to the CSR. The kon's for isolated CSR and HSPG, and the koff for isolated HSPG, did not differ significantly from those for intact cells konC = 2.50 x 10(8) M-1 min-1; konH = 0.92 x 10(8) M-1 min-1; koffH = 0.095 min-1). However, the off rate for isolated CSR (koffC = 0.048 min-1) was statistically indistinguishable from the off rate for HSPG and 16-fold greater than the off rate for CSR on intact cells. The "high-affinity" binding of bFGF to intact cells probably refers only to a complex of bFGF with both CSR and HSPG, and not to the CSR alone.     

Antisense oligonucleotides to crabp I and II alter the expression of TGF-β3, RAR-β, and tenascin in primary cultures of embryonic palate cells

Summary The cellular retinoic acid-binding proteins (CRABPs) are thought to modulate the responsiveness of cells to retinoic acid (RA). We have previously shown that primary cultures of murine embryonic palate mesenchymal (MEPM) cells express both CRABP-I and CRABP-II genes and that this expression is regulated by RA and transforming growth factor β (TGF-β). These cells also express high levels of TGF-β3, which is also regulated by RA and TGF-β. We have used an antisense strategy to investigate the role of the CRABPs in retinoid-induced gene expression. Subconfluent cultures of MEPM cells were treated for several days with phosphorothioate modified 18-mer oligonucleotides antisense to CRABP-I or CRABP-II and then with all-trans-retinoic acid at a concentration of 3.3 μM or 0.33 μM for 5 or 22 h. Total RNA was then extracted and the expression of TGF-β3, retinoic acid receptor β (RAR-β), and tenascin was assessed by northern blot analysis. Antisense oligonucleotides to CRABP-I partially inhibited the RA-induced TGF-β3, RAR-β, and tenascin mRNA expression. The corresponding mis-sense oligonucleotides were without effect. Antisense oligonucleotides to CRABP-II also partially inhibited RA-induced expression of these genes. As with the CRABP-I antisense, mis-sense oligonucleotides to CRABP-II had no effect. These data suggest that both CRABPs modulate the responsiveness of MEPM cells to retinoic acid. Inhibition of endogenous CRABP expression renders MEPM cells less responsive to RA with respect to induction of TGF-β3, RAR-β, and tenascin gene expression. These results have important implications for our understanding of the role of the CRABPs in retinoid teratology.     

Building the State, Making the Nation: The Bases and Limits of State Centralization in "Modern" Peru

Most contemporary discussions of the nature of the state, state-building, and national consciousness formation are based on an oppositional model of state-society relations. In such models, state-building is depicted as a process in which the state must impose its central institutions and cultural/moral values on the recalcitrant local populations found within its territorial boundaries who, in resisting the state, cling to a myriad of local, oppositional identities. The present article critiques the oppositional model, proposes an alternative conceptualization that encompasses dimensions of cooperation and conflict in state-society relations, and thus points toward the forces that underlie the fragmentation and consolidation of states and national cultures.     

Spectroscopic studies of the manganese complex of Photosystem II

Our recent EPR and EXAFS experiments investigating the structure of the oxygen-evolving complex of PS II are discussed. PS II treatments which affect the cofactors calcium and chloride have been used to poise samples in modified forms of the S-states, S₁, S₂ and S₃. X-ray absorption studies indicate a similar overall structure for the manganese complex between treated and native samples although the influence of the treatments and cofactors is observed. Manganese oxidation (or oxidation of a ligand to the manganese cluster) is indicated to occur on each of the transitions S₁ S₂ and S₂ S₃ in these modified samples. The cluster appears to contain at least two inequivalent Mn-Mn pairs. In the native samples the Mn-Mn distance is 2.7 Å, but in samples where the calcium site is affected, one of the pairs has a 3.0 Å Mn-Mn distance. The intensity of the 3.3/3.6 Å interaction is reduced on sodium chloride treatment (calcium depletion) perhaps indicating calcium binding close to the manganese cluster. From EPR data we also propose that treatments which affect calcium and chloride binding cause a modification of the native S₂ state, slow the reduction of Y_z and allow an S3 EPR signal to be observed following illumination. The origin of the S3 EPR signal, a modified S₃ or S₂ X where X is an organic radical of unknown charge, is discussed in relation to the results from the EXAFS studies.Abbreviations EPR electron paramagnetic resonance spectroscopy - EXAFS extended X-ray absorption fine structure - HTG n-heptyl -d-thioglucoside - MES 2(N-morpholino)ethanesulfonic acid - OEC oxygen evolving complex - PPBQ phenyl-1,4-benzoquinone - PS II Photosystem II - Y_z redox active tyrosine