Characterization of the cell division factor ZapB of Escherichia coli

PhD ThesisBacterial cell division relies on the formation and contraction of the Z-ring, coordinated and regulated by a dynamic protein complex called divisome. Here, we show that ZapB, a newly identified cell division factor in Escherichia coli, is recruited to the division site in the early stages of Z-ring assembly by ZapA. Inactivation or overproduction of ZapA caused ZapB delocalization and diffusion. Bacterial two-hybrid and in vitro assays showed that ZapB interacts directly with ZapA and, through it, with FtsZ and that the three proteins together can form a highmolecular- weight complex. Furthermore, during the cell cycle ZapB closely followed FtsZ dynamic localization but interestingly, using high-resolution 3D reconstruction microscopy, we found that it formed a ring located on the inside of the Z-ring, consistently in all cells in consecutive cell division events. Only in the absence of the bacterial actin homologue MreB, ZapB was not able to constrict ahead of FtsZ and instead co-localized seemingly perfectly with the Z-ring. Morphological analysis of cells carrying a zapB deletion and the ftsZ84 allele exhibited a synthetic detrimental phenotype and cell division defects. A model in which ZapB further increases the lateral association of FtsZ filaments by cross-linking ZapA molecules bound to adjacent FtsZ filaments is supported by light scattering assays and analysis of structures formed by FtsZ-ZapA-ZapB using electron microscopy. Surprisingly, ZapB seemed to be active in cell division in the absence of ZapA raising the possibility that ZapB might have a secondary ZapA-independent function

PDMS-based films containing surface-active amphiphilic block copolymers to combat fouling from barnacles B. amphitrite and B. improvisus

Surface-active amphiphilic diblock copolymers, Si-EFS14 and Si-EFS71, consisting of a poly(dimethyl siloxane) block (degree of polymerisation 11) and a poly(4-(triethyleneglycol monomethyl ether)-2,3,5,6-tetrafluorostyrene) block (average degree of polymerisation 14 and 71) were synthesised by ATRP. Films were prepared by incorporating each copolymer (4 wt%) into a PDMS matrix, which was then condensation cured. Bioassays were performed on the films using two barnacle species, Balanus amphitrite and Balanus improvisus, at different stages of their life cycles. The cyprids of B. improvisus settled on all test surfaces in higher numbers than those of B. amphitrite. However, the juveniles of B. improvisus were more easily removed from the films containing the copolymer Si-EFS14 than from those containing the copolymer Si-EFS71. An XPS analysis revealed that the near-surface region of copolymer Si-EFS71 was enriched in oxyethylenic chains and became even more populated by these hydrophilic chains after the films were immersed in water

Julolidine fluorescent molecular rotors as vapour sensing probes in polystyrene films

We introduce a new sensing polymer system for detection of volatile organic compounds (VOCs) based on the optical response of polystyrene (PS) films doped with julolidine fluorescent molecular rotors (FMRs). The julolidine FMRs exhibited viscosity-dependent changes in the fluorescence intensity, that was enhanced when glycerol was added to ethanol solutions and when they were dispersed in PS films. Thus, reduction in medium mobility slowed down internal motions and allowed for a major radiative decay pathway. The FMR/PS films were exposed to several VOCs, and showed a significant decrease in fluorescence emission when exposed to chloroform, whereas a negligible variation in their emission occurred when methanol was utilized. This vapour sensing behaviour was much more evident when a perfluorodecyl chain was linked to the julolidine core being the molecule segregated at the film surface. This responsive behaviour was affected by solvent composition and its reproducible response was easily determined by luminescence experiments

Experiences from a winter school on landscape agronomy: Stakes, difficulties, perspectives

International audienceIn the latest fifteen years, agronomic research has shown a growing interest for studies which link farm or field scale to landscape scale. Thus, agronomy is called to renew its research questions and methodologies, and as well its educational programmes. In this context, some French and Italian researchers interested in these topics, coming from different scientific fields but sharing interests on landscape scale issues in research and higher education, decided to join their efforts around a common one-week educational programme on Landscape Agronomy for undergraduate and PhD students. Their aim has been to develop a new form of knowledge transfer and application on Landscape Agronomy approaches to students of SSSA-Pisa (IT) and of the PhD School of ABIES-AgroParisTech-Paris (FR). The educational programme consisted of three phases : 1) some theoretical contributions supported by presentations on : issues regarding agriculture and farming practices at landscape level, main approaches on environmental functions of agriculture, changes in farmers practices driven by environmental questions,spatial organization of agricultural activities, role of farming in ecological dynamics, identification of complementarities among agro-environmental functions, environmental impacts of cropping systems,biodiversity influence on agro-ecosystem functions and vice-versa ; 2) two case-studies: a macro level one(at landscape scale) to analyse the role of agriculture on landscape dynamics, and a micro level one (at farm scale) to analyse farming practices and their environmental impacts ; 3) a final evaluation of the educational programme based on : contents of oral presentations on fieldwork results, global evaluation of the educational programme contents by all the participants, each individual ex-post analysis of fieldwork results. The evaluation of the students and teachers underlines benefits, requests and perspectives for education in landscape agronomy. Furthermore, this experience stimulated a collective conceptual and methodological debate that confirmed the necessity to favour and organise experience exchanges on researching and learning in landscape agronomy

Tetranuclear coordination assemblies based on half-sandwich ruthenium(II) complexes. Non covalent binding to DNA and cytotoxicity.

The reaction of [(cymene)RuCl2]2 with K2Hoxonate (H3oxonic = 4,6-dihydroxy-2-carboxy-1,3,5-triazine acid) in methanol leads to the formation of the dinuclear half-sandwich ruthenium(II) complex [(cymene)2Ru2(\u3bc-Hoxonato)Cl2] (1a). Removal of the chloride ligands of 1a by treatment with AgCF3SO3 yields [(cymene)2Ru2(\u3bc-Hoxonato)(CF3SO3)2] (1b), which, upon posterior reaction with N,N'-linkers (L = 4,4'-bipyridine (4,4'-bpy), 4,7-phenantroline (4,7-phen)), gives rise to the formation of the tetranuclear open boxes [(cymene)4Ru4(\u3bc-Hoxonato)2(\u3bc-N,N'-L)2](CF3SO3)4 (2a, L = 4,4'-bpy; 2b, L = 4,7-phen). These systems have been characterized by 1HNMR, UV-vis, and ESI-MS. The single-crystal structures of the dinuclear precursor 1a and of the clathrate 2b 824,7-phen have been determined. The interaction of these systems with cysteine, mononucleotides, and calf-thymus DNA has been studied by means of 1HNMR, UV-vis, circular dicroism, competitive binding assays, and atomic force microscopy imaging. The results show that the robust tetracationic ruthenium(II) cyclic systems 2a and 2b do not give ligand exchange reactions toward biorelevant ligands. Nevertheless, these systems are able to noncovalently bind to DNA, probably at the surface of the major groove, inducing significant conformational changes in this biomolecule. It is also interesting to note that compounds 2a and 2b, in spite of only giving supramolecular interactions with biomolecules, exhibit antitumor activity, particularly toward the human ovarian cancer cell line A2780cisR, showing acquired resistance to cisplatin, with respective 4.6 and 8.3 microM IC50 values

Cartografando modos de trabalhar e modos de subjetivar de um órgão público

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Amphiphilic pentablock copolymers and their blends with PDMS for antibiofouling coatings

Well-defined amphiphilic pentablock copolymers Siy-(EGx-FAz)2 composed of polysiloxane (Si), polyethylene glycol (EG), and perfluorohexylethyl polyacrylate (FA) blocks are synthesized by ATRP of FA monomer starting from a difunctional bromo-terminated macroinitiator. Diblock copolymers EGx-FAz are also synthesized as model systems. The block copolymers are used, either alone or blended with a PDMS matrix in varied loadings, to prepare antibiofouling coatings. Angle-resolved XPS and contact angle measurements show that the coating surface is highly enriched in fluorine content but undergoes reconstruction after contact with water. Protein adsorption experiments with human serum albumin and calf serum highlight that diblock copolymers resist protein adhesion better than do pentablock copolymers. Blending of the pentablock copolymer with PDMS results in increased protein adsorption. By contrast, the PDMS-matrix coatings show high removal percentages of sporelings of the green fouling alga Ulva linz

Cerebrospinal fluid anti-Epstein-Barr virus specific oligoclonal IgM and IgG bands in patients with clinically isolated and Guillain-Barré syndrome

Epstein-Barr virus (EBV) has been implicated in multiple sclerosis (MS) pathogenesis. We aimed to assess the frequency of EBV-specific IgG and IgM oligoclonal bands (OCB) in cerebrospinal fluid (CSF) of 50 patients with clinically isolated syndrome (CIS) and in 27 controls with Guillain-Barré syndrome (GBS). Furthermore, we assessed correlations between the presence of OCB and CIS patients' CSF, MRI, and clinical variables. There was no difference in the proportion of CIS and GB patients with positivity for anti-EBV-specific IgG/IgM OCB. There were no correlations between OCB and analyzed variables, nor were they predictive of a higher disability at 3 years

Amphiphilic modified-styrene copolymer films: Antifouling/fouling release properties against the green alga Ulva linza

Surface-active copolymers of a styrene carrying a polysiloxane side chain (SSi) and a triethyleneglycol monomethyl ether-modified pentafluorostyrene (EFS) (39 and 77 mol% EFS) were prepared and incorporated (8 wt% loading) into a polydimethyl siloxane (PDMS) matrix to produce crosslinked blend films. The wettability of the surface-active copolymer films and PDMS-blend films was investigated by contact angle measurements. An angle-resolved X-ray photoelectron spectroscopy (XPS) of the surface chemical composition before and after immersion in water for 7 days enabled location of the hydrophilic oxyethylenic segments of EFS within the top 10 nm from the film surface. Laboratory bioassays on the blend films against the marine green alga Ulva linza evidenced that the films containing the copolymer with the larger EFS content showed greater resistance to settlement of zoospores of U. linza, whereas both films had superior fouling-release properties of sporelings (young plants) compared to the PDMS standard films

Replication termination without a replication fork trap

International audienceBacterial chromosomes harbour a unique origin of bidirectional replication, oriC. They are almost always circular, with replication terminating in a region diametrically opposite to oriC, the terminus. The oriC-terminus organisation is reflected by the orientation of the genes and by the disposition of DNA-binding protein motifs implicated in the coordination of chromosome replication and segregation with cell division. Correspondingly, the E. coli and B. subtilis model bacteria possess a replication fork trap system, Tus/ter and RTP/ter, respectively, which enforces replication termination in the terminus region. Here, we show that tus and rtp are restricted to four clades of bacteria, suggesting that tus was recently domesticated from a plasmid gene. We further demonstrate that there is no replication fork system in Vibrio cholerae, a bacterium closely related to E. coli. Marker frequency analysis showed that replication forks originating from ectopic origins were not blocked in the terminus region of either of the two V. cholerae chromosomes, but progressed normally until they encountered an opposite fork. As expected, termination synchrony of the two chromosomes is disrupted by these ectopic origins. Finally, we show that premature completion of the primary chromosome replication did not modify the choreography of segregation of its terminus region