Erratum to: Exogenous Adipokine Peptide Resistin Protects Against Focal Cerebral Ischemia/Reperfusion Injury in Mice

Neurochemical Research -     

A new role for PGRP-S (Tag7) in immune defense: lymphocyte migration is induced by a chemoattractant complex of Tag7 with Mts1

PGRP-S (Tag7) is an innate immunity protein involved in the antimicrobial defense systems, both in insects and in mammals. We have previously shown that Tag7 specifically interacts with several proteins, including Hsp70 and the calcium binding protein S100A4 (Mts1), providing a number of novel cellular functions. Here we show that Tag7–Mts1 complex causes chemotactic migration of lymphocytes, with NK cells being a preferred target. Cells of either innate immunity (neutrophils and monocytes) or acquired immunity (CD4⁺ and CD8⁺ lymphocytes) can produce this complex, which confirms the close connection between components of the 2 branches of immune response.     

Oxidative bioconversion of toluene to 1,3-butadiene-1,4-dicarboxylic acid (cis,cis-muconic acid)

Pseudomonas sp. strains, isolated from soil, utilized toluene as their sole carbon source through ameta cleavage pathway. Strains metabolizing toluene through anortho cleavage pathway were selected from the wild typemeta strain. Theortho pathway strains were subjected to chemostat selection to obtain a fast-growing strain with doubling time reduced from 14 to 1.2 h. Benzoale and antibiotics enrichment selection procedures were utilized to select a blocked mutant. The blocked mutant grew on acetate as its sole carbon source and oxidatively converted toluene tocis, cis-muconic acid. Double-blocked and muconate-permeable mutants were also selected to reduce reversion frequency and to enhance muconic acid production. In shake-flask experiments, muconic acid at 3.5 g/l was obtained after 2 days of fermentation. In a 14 l fermenter, muconic acid was produced at 45 g/l in 4 days of controlled fed-batch fermentation. The oxidative bioconversion process was also demonstrated in a 1500 l fermenter.

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Résumé Des souches dePseudomonas sp., isolées du sol, ont utilisé le toluène comme seule source de carbone par la vole de la rupture de cycle enmeta. On a sélectionné des souches métabolisant le toluène par la voie de la rupture de cycle enortho, à partir de la souche sauvage de typemeta. Les souches de la voieortho ont été soumises à la sélection en chémostat pour obtenir des souches à croissance rapide dont le temps de doublement est rédult de 14 à 1.2 h. Les procédures de sélection par enrichissement sur benzoale et antibiotiques ont été utilisées pour sélectionner un mutant bloqué. Le mutant bloqué croît sur acétate comme seule source de carbone et convertit le toluène par voie oxydative en acidecis,cis-muconique. On a également sélectionné des mutants doublement bloqués et perméables au muconate pour réduire la lréquence de réversion et pour augmenter la production d'acide muconique. En expérimentation en flacons agités, on a obtenu 3.5 g/l d'acide muconique après 2 jours de fermentation. En fermentuer de 14 l, on a produit 45 g/l d'acide muconique en 4 jours de fermentation contrôlée en milieu non renouvelé à allmentation étagée. Le processus de bioconversion oxydative a également été démontré en fermenteur de 1500 l.

     

Cell-specific expression of pyruvate, pi dikinase : in situ mRNA hybridization and immunolocalization labeling of protein in wheat seed

Pyruvate, Pi dikinase (PPDK) is a key enzyme in the C4 photosynthetic pathway. However, its metabolic role in C3 plants remains uncertain. Northern blot analyses of PPDK mRNAs from wheat leaves and seeds probed with maize PPDK cDNA indicates the presence of organ-specific mRNAs. Immunofluorescent labeling of protein in wheat seed demonstrate that the PPDK polypeptide and the ribulose-1, 5-bisphosphate carboxylase small subunit polypeptide are localized predominantly in the aleurone layer and the chlorophyllous pericarp tissue, respectively. This differential distribution of the two polypeptides in wheat seed is paralleled by the differential localization of the their mRNAs as revealed by in situ hybridization. These results suggest a distinct role of cytoplasmic PPDK in seeds, which is different from the well established role in C4 photosynthesis.     

Dominant pleiotropy controls enzymes co-segregating with paraquat resistance in Conyza bonariensis

Summary The genetics of paraquat-resistance in Conyza bonariensis was studied. Reciprocal crosses were prepared between resistant and sensitive individuals. The enzymes of the pathway that detoxifies superoxide to innocuous oxygen species involved in resistance were evaluated in the F₁ and F₂ generations. All F₁ plants were as resistant as the resistant parent, irrespective of parental sex, demonstrating dominance and excluding maternal inheritance. The activities of superoxide-dismutase, ascorbate-peroxidase and glutathione-reductase in the F₁ were constitutively as high as in the resistant parent. Resistance in the F₂ generation was distributed in a 31 ratio (resistant to sensitive). Leaves from F₂ plants were removed for a resistance assay and enzyme immuno-assays of single plants were performed. The high levels of superoxide-dismutase and glutathione-reductase, the two enzymes for which antibodies were available, were similar in resistant individuals to the levels in the resistant parent; the levels were low in the susceptible individuals. These results indicate either a very tight linkage, or more probably, that one dominant nuclear gene controls resistance by pleiotropically controlling the levels of enzymes of the activeoxygen detoxification pathway.     

Cloning of plant cDNAs encoding calmodulin-binding proteins using35S-labeled recombinant calmodulin as a probe

Radiolabelled calmodulin has previously been used to screen cDNA expression libraries to isolate calmodulin-binding proteins. We have modified this technique for the isolation of plant calmodulin-binding proteins. [³⁵S]-methionine was used instead of the inorganic [³⁵S]-sulfate, or¹²⁵I used in previous methods. In addition, theE. coli pET expression system was chosen to obtain high levels of recombinant calmodulin at the time of labelling. The procedure thus takes into account both the specific activity of the probe and the amount of protein necessary for screening a large number of filters. Here we describe in detail a procedure for the production and purification of [³⁵S]-recombinant calmodulin and the use of the radiolabelled protein as a probe to screen plant cDNA expression libraries. The [³⁵S]-labeled calmodulin probe easily detects the λICM-1 phage encoding a partial mouse calmodulin-dependent protein kinase II that was previously isolated using a [¹²⁵I]-calmodulin probe (Sikela and Hahn, 1987). Subsequently, a tobacco root cDNA expression library was screened and a positive clone encoding a calcium-dependent calmodulin-binding protein was isolated.