Roles of curli,cellulose and BapA in <Emphasis Type="Italic">Salmonella</Emphasis> biofilm morphology studied by atomic force microscopy

Background  

Curli, cellulose and the cell surface protein BapA are matrix components in Salmonella biofilms. In this study we have investigated the roles of these components for the morphology of bacteria grown as colonies on agar plates and within a biofilm on submerged mica surfaces by applying atomic force microscopy (AFM) and light microscopy.     

Identification of AtPIS, a phosphatidylinositol synthase from Arabidopsis.

Phosphatidylinositol synthase is the enzyme responsible for the synthesis of phosphatidylinositol, a key phospholipid component of all eukaryotic membranes and the precursor of messenger molecules involved in signal transduction pathways for calcium-dependent responses in the cell. Using the amino acid sequence of the yeast enzyme as a probe, we identified an Arabidopsis expressed sequence tag potentially encoding the plant enzyme. Sequencing the entire cDNA confirmed the homology between the two proteins. Functional assays, performed by overexpression of the plant cDNA in Escherichia coli, a bacteria which lacks phosphatidylinositol and phosphatidylinositol synthase activity, showed that the plant protein induced the accumulation of phosphatidylinositol in the bacterial cells. Analysis of the enzymatic activity in vitro showed that synthesis of phosphatidylinositol occurs when CDP-diacylglycerol and myo-inositol only are provided as substrates, that it requires manganese or magnesium ions for activity, and that it is at least in part located to the bacterial membrane fraction. These data allowed us to conclude that the Arabidopsis cDNA codes for a phosphatidylinositol synthase. A single AtPIS genetic locus was found, which we mapped to Arabidopsis chromosome 1.     

A Plastidial Lysophosphatidic Acid Acyltransferase from Oilseed Rape

The biosynthesis of phosphatidic acid, a key intermediate in the biosynthesis of lipids, is controlled by lysophosphatidic acid (LPA, or 1-acyl-glycerol-3-P) acyltransferase (LPAAT, EC 2.3.1.51). We have isolated a cDNA encoding a novel LPAAT by functional complementation of the Escherichia coli mutant plsC with an immature embryo cDNA library of oilseed rape (Brassica napus). Transformation of the acyltransferase-deficient E. coli strain JC201 with the cDNA sequence BAT2 alleviated the temperature-sensitive phenotype of the plsC mutant and conferred a palmitoyl-coenzyme A-preferring acyltransferase activity to membrane fractions. The BAT2 cDNA encoded a protein of 351 amino acids with a predicted molecular mass of 38 kD and an isoelectric point of 9.7. Chloroplast-import experiments showed processing of a BAT2 precursor protein to a mature protein of approximately 32 kD, which was localized in the membrane fraction. BAT2 is encoded by a minimum of two genes that may be expressed ubiquitously. These data are consistent with the identity of BAT2 as the plastidial enzyme of the prokaryotic glycerol-3-P pathway that uses a palmitoyl-ACP to produce phosphatidic acid with a prokaryotic-type acyl composition. The homologies between the deduced protein sequence of BAT2 with prokaryotic and eukaryotic microsomal LAP acytransferases suggest that seed microsomal forms may have evolved from the plastidial enzyme.     

Fracture de verge: à propos de 12 cas

The authors would like to report their management of 12 patients with penile fracture. Their protocol consisted of emergency surgical repair, which gave the patients the greatest chance of full recovery, i.e. normal erection and effective sexual intercourse. A review of the literature shows the relative rarity of this traumatic disease and emphasizes the value of cavernosography, MRI and sonography in the diagnosis.     

A potent antimicrobial protein from onion seeds showing sequence homology to plant lipid transfer proteins.

An antimicrobial protein of about 10 kD, called Ace-AMP1, was isolated from onion (Allium cepa L.) seeds. Based on the near-complete amino acid sequence of this protein, oligonucleotides were designed for polymerase chain reaction-based cloning of the corresponding cDNA. The mature protein is homologous to plant nonspecific lipid transfer proteins (nsLTPs), but it shares only 76% of the residues that are conserved among all known plant nsLTPs and is unusually rich in arginine. Ace-AMP1 inhibits all 12 tested plant pathogenic fungi at concentrations below 10 micrograms mL-1. Its antifungal activity is either not at all or is weakly affected by the presence of different cations at concentrations approximating physiological ionic strength conditions. Ace-AMP1 is also active on two Gram-positive bacteria but is apparently not toxic for Gram-negative bacteria and cultured human cells. In contrast to nsLTPs such as those isolated from radish or maize seeds, Ace-AMP1 was unable to transfer phospholipids from liposomes to mitochondria. On the other hand, lipid transfer proteins from wheat and maize seeds showed little or no antimicrobial activity, whereas the radish lipid transfer protein displayed antifungal activity only in media with low cation concentrations. The relevance of these findings with regard to the function of nsLTPs is discussed.     

Turning, compacting and the addition of water as factors affecting gaseous emissions in farm manure composting

Composting allows simple management of animal manure but excessive aeration can increase emissions of polluting gases such as ammonia or nitrous oxide. The aim of the present work was to determine the effect of three techniques--turning, compacting and the addition of water--on gaseous emissions. One ton of cattle manure and 3 tons of turkey manure were composted in two and four cells for 46 and 51 days respectively. The manure was either turned, wetted, or compacted. Emissions of carbon dioxide, water vapor, ammonia and nitrous oxide were monitored. The results show that turning did not alter the free air space. Compacting can be used specifically to reduce the water loss. A reduction of free air space by 20-60%, either by compacting or adding water (or both), reduced the ammonia and nitrous oxide emissions by 30-70%.     

新疆杏树吐伦球坚蚧的防治指标与田间药效试验

通过对吐伦球坚蚧Rhodococcus turanicus Arch.种群密度与杏树产量的调查和田间药效试验研究得出:40％毒死蜱·机油乳剂对其防治效果较好,7d后的防治效果可达83.99％;初步探明了吐伦球坚蚧的为害损失率,建立了杏树损失率与吐伦球坚蚧种群密度的回归模型为y=24.671Ln (x)-16.56(y为...     

Composition of organic matter in sandy relict and cultivated heathlands as examined by pyrolysis-field ionization MS

Unusually high SOC levels have been reported for sandy cropland soils in North-Western Europe. A potential link with their general heathland land-use history was investigated by comparing two soil pairs of relict heathland and cultivated former heathland in the Belgian sandy region. A sequential chemical fractionation yielded similar sizes in corresponding SOM fractions between the heathland and cropland soils (i.e. NaOCl resistant: 12.3–15.0 g C kg⁻¹ and NaOCl + HF resistant: 2.6–5.3 g C kg⁻¹). Higher amounts of clay sized N in the cropland plots can be attributed to N additions from mineral fertilizers and animal manure. Temperature resolved Pyrolysis Field Ionization Mass Spectroscopy analysis showed that the composition of both relict heathland and cultivated soils was surprisingly similar, in spite of over 60 years of intense cropland management. The mass spectra of SOM in both heathland-cropland soil pairs investigated was dominated by signals from lipids, alkylaromatics and sterols. The accumulation of this SOM rich in aliphatics was logically linked to the high input of lipids, long-chain aliphatics and sterols from heathland vegetation and the low soil pH and microbial activity. Based on the relatively high OC surface loadings of HF-extractable OM (13–44 mg C m⁻² Fe and 1.2–2.3 mg C m⁻² clay), direct organo-mineral bonds between OM and Fe-oxides or clay minerals seem to be only partly involved as a stabilization mechanism in these soils. The distinct bimodal shape of the thermograms indicates that OM-crosslinking could furthermore contribute substantially to SOM stabilization in these soils. This study therefore corroborates the previously proposed view that lipids may be bound in networks of alkylaromatics, the structural building blocks of OM macromolecules. We hypothesize that such binding is able to explain the measured retention of these OM components, even under several decades of cropland management.     

Direct interaction between CCN family protein 2 and fibroblast growth factor 1

In an attempt to find out a new molecular counterpart of CCN family protein 2 (CCN2), a matricellular protein with multiple functions, we performed an interactome analysis and found fibroblast growth factor (FGF) -1 as one of the candidates. Solid-phase binding assay indicated specific binding between CCN2 and FGF-1. This binding was also confirmed by surface plasmon resonance (SPR) analysis that revealed a dissociation constant (Kd) of 3.98 nM indicating strong molecular interaction between the two. RNA analysis suggested that both FGF-1 and CCN2 could be produced by chondrocytes and thus their interaction in the cartilage is possible. These findings for the first time indicate the direct interaction of CCN2 and FGF-1 and suggest the co-presence of these molecules in the cartilage microenvironment. CCN2 is a well-known promoter of cartilage development and regeneration, whereas the physiological and pathological role of FGF-1 in cartilage mostly remains unclear. Biological role of FGF-1 itself in cartilage is also suspected.