Targeting carbohydrates: a novel paradigm for fungal control

The tendentious decrease in global arable land and the fast increment in world population will inevitably lead to a critical point. It is therefore imperative to protect crops from pathogen attack. Their deleterious impact on human health is not better, with several invasive infections reaching mortality rates up to 80%. The door is definitely open for biological control, based on molecular interactions between host and pathogen. The remarkable coding capacity of oligosaccharides, far superior to those of nucleotides and amino acids, justifies the emergence of the sugar code as the third life code. The word exoglycome is proposed to represent the oligosaccharide side chains presented at the outer surface of cell membranes, thus excluding the cell wall carbohydrates. The exoglycome plays a fundamental role in cell-cell recognition, in distinction between self from non-self, in the warfare between host and pathogen before infection is established, and in certain diseases such as cancer. Unlike pathogen cell wall carbohydrates, the exoglycome has been neglected as a target for developing effective ways to control disease. Pathogens often induce changes in their cell wall and/or exoglycome to elude and/or circumvent host defences. Identification of such host-induced alterations may prove extremely important for both treatment and diagnosis. Reading the sugar-encoded messages is mainly played by a class of carbohydrate-binding receptors called lectins. The available evidence strongly suggests that a large number of lectins remain to be discovered. Their role in defence should not be underestimated. A high-throughput procedure capable of single-step isolation of novel lections from previously unknown biological samples has recently been developed.     

Organoarsenical species contents in fresh and processed seafood products

A study was carried out to determine organic species of arsenic in the main varieties of seafood consumed in the Basque country (Spain). The concentrations of arsenobetaine (AB), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA), arsenocholine (AC), and tetramethylarsonium ion (TMA(+)) in 64 samples corresponding to different food items are presented. The study provides information about a possible distribution pattern of organoarsenical species in seafood products. AB was detected in all of the samples [0.3-104.1 microg g(-1) dry weight (dw)]. DMA was detected in all of the samples except squid and salted cod (0.027-1.757 microg g(-1) dw). MMA was detected only in certain fatty fish (0.004-0.028 microg g(-1) dw) and bivalves (0.031-0.047 microg g(-1) dw). AC was only present in some samples of lean fish (0.014-0.089 microg g(-1) dw), and TMA(+) was detected only in anchovy (0.039-0.169 microg g(-1) dw) and crustaceans (0.044-0.966 microg g(-1) dw).     

Spatio-temporal variations in nutrient concentration in soil solution under greenhouse tomato

The aim of this trial was to study the spatio-temporal variability in solution nutrient concentration under intensive greenhouse tomato production, to determine the number of suction-cups needed to obtain a representative sample and the influence by the position in the greenhouses. Twenty sampling points were selected within the greenhouse with one suction-cup per sampling point. One soil solution were sampled per point at weekly intervals to analyze for pH, electrical conductivity, chloride, nitrate, phosphate, sulfate, sodium, potassium, calcium, and magnesium (EC, Cl^?, NO₃^?, H₂PO₄^?, SO₄^2—, Na⁺, K⁺, Ca²⁺, and Mg²⁺) concentrations. The pH, Cl^?, H₂PO₄^?, and SO₄^2? concentrations showed no spatio-temporal variation but EC, NO₃^?, and K⁺ showed temporal variation. The spatial variability in EC, K⁺_, Na⁺, Mg²⁺, and Ca²⁺ can be influenced by microclimate and topography. The numbers of suction cups required for a representative sample ranged from 1 to 10 depending on nutrient.     

Conjugates of bovine serum albumin with chitin oligosaccharides prepared through the Maillard reaction

Chitin neoglycoconjugates (BSA-CO) were obtained by the conjugation of bovine serum albumin (BSA) with chitin oligosaccharides (CO) through the Maillard reaction (nonenzymatic glycation). CO produced by acid hydrolysis of chitin were fractionated using an ultrafiltration membrane system (1-3 kDa cutoff). The Maillard reaction was carried out by heating a freeze-dried mixture containing BSA and CO at 60 °C (under 43% relative humidity for 6 and 12 h). BSA-CO were characterized by available amino groups content, intrinsic tryptophan emission spectra, gel electrophoresis, and mass spectrometry. Biological assays included interaction with wheat germ agglutinin (WGA) and with bacterial adhesins of Escherichia coli K88+ and Salmonella choleraesuis. Glycation of BSA was revealed by reduction of available amino groups and fluorescence intensity and also retarded migration through SDS-PAGE. Conjugation of BSA with chitin oligomers appeared to be time dependent and was confirmed by mass spectrometry, by which molecular mass increase for monomers and dimers was observed. Monomers were estimated to contain either one or two glycation sites (at 6 and 12 h of treatment, respectively), with one or two tetrasaccharide units attached. Consequently, dimers showed two or four glycation sites. BSA-CO presented biological recognition by WGA and E. coli K88+ and S. cholerasuis adhesins. The strategy used in this work represents a simple method to obtain glycoconjugates to study applications involving protein-carbohydrate recognition.     

Putative Role of <Emphasis Type="Italic">Flavobacterium</Emphasis>, <Emphasis Type="Italic">Dokdonella</Emphasis> and <Emphasis Type="Italic">Methylophilus</Emphasis> Strains in Paracetamol Biodegradation

Paracetamol, the most widely and globally used analgesic and antipyretic, is easily accumulated in aquatic environments. In the present study, the biodegradation of paracetamol in different media (one for general growth, one specific for sulfate reducing bacteria, a mineral salts medium and municipal wastewater) inoculated with two types of sludge (from anaerobic lagoon and from oxidation ditch) under different oxygenic conditions (anoxic; moderate oxygenation in open flasks and high oxygenation by aeration) was investigated. In addition, bacteria with relative abundances increasing simultaneously with paracetamol degradation, when this drug was the only carbon source, thus with a putative role in its degradation, were identified using 16S rRNA gene sequences. The results show that aerobic microorganisms had a major role in the degradation of paracetamol, with 50 mg/L totally removed from municipal wastewater after 2 days incubation with aeration, and that the metabolites 4-aminophenol and hydroquinone plus one compound not identified in this work were produced in the process. The identification of bacteria with a role in the degradation of paracetamol revealed a strain from genus Pseudomonas with the highest final relative abundance of 21.2%, confirming previous works reporting strains of this genus as paracetamol decomposers. Besides, genera Flavobacterium, Dokdonella and Methylophilus were also in evidence, with initial relative abundances of 1.66%, 1.48 and 0.00% (not detected) in the inoculum and 6.91%, 3.80 and 3.83% after incubation, respectively. Therefore, a putative role of these genera in paracetamol biodegradation is suggested for the first time.

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Graphical Abstract ?

     

Evidence that fresh weight measurement is imprecise for reporting the effect of plant growth-promoting (rhizo)bacteria on growth promotion of crop plants

Eight greenhouse experiments were performed to compare the effect of seven plant growth-promoting (rhizo)bacteria (PGPR/PGPB) on fresh and dry weights of four crop plants. This has been done to validate if fresh weight measurements of plant variables can serve as reliable values when reporting the effect of these bacteria on plant growth. These experiments show that the growth promotion effects by the tested PGPR/PGPB, including Bacillus amyloliquefaciens GB03, Bacillus subtilis IN-937B, Bacillus altitudinis INR7, and Pseudomonas mandelii 89B-27 in corn and cucumber and Azospirillum brasilense Cd, A. brasilense Sp 245, and Azospirillum lipoferum Br 17 in pepper and tomato, varied significantly between fresh and dry weights of shoot, root, and/or whole plant in the repeated greenhouse experiments. These results support our hypothesis that using fresh weight determination for assessing plant growth promotion by beneficial bacteria is inherently faulty. Therefore, it is recommended that dry weight determination rather than fresh weight determination is used for plant growth promotion tests.     

Manganese Supply and pH Influence Growth,Carboxylate Exudation and Peroxidase Activity of Ryegrass and White Clover

ABSTRACT

The effects of differential manganese (Mn) supply (0 to 355 μ M) and pH (4.8 and 6.0) on dry weight (DW), tissue concentrations of Mn, exudation of carboxylates, and the peroxidase activity were studied in ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.) grown in nutrient solution. In both plant species, the increase in Mn supply caused a significant reduction in DW due to severe Mn toxicity, especially at pH 4.8. The critical toxicity concentration of Mn in shoots was 421 mg kg^{? 1} for ryegrass and 283 mg kg^{? 1} for white clover. For both plant species, an increase in Mn supply levels stimulated the exudation of carboxylates and the activity of peroxidase, which was related to stress conditions. The highest amount of carboxylates was exuded at pH 4.8. There was no clear effect of carboxylates on the complexation of Mn^{2 +}.     

Detection of two minor phosphorylation sites for bovine κ-casein macropeptide by reversed-phase liquid chromatography-tandem mass spectrometry

This work addresses the characterization of phosphopeptides in bovine κ-casein macropeptide by reversed-phase liquid chromatography-electrospray ionization-tandem mass spectrometry (RPLC-ESI-MS(2)). Two different mass spectrometers, equipped with an ion trap (IT) or a quadrupole time-of-flight (Q-TOF) analyzer, were used to perform an accurate phosphorylation site assignment. A total of 8 phosphopeptides from 26 identified peptides were characterized. MS(2) spectra of phosphopeptides were dominated by the neutral loss of a phosphoric acid molecule (H(3)PO(4)) and sufficient informative fragment ions resulting from peptide backbone cleavages enabling the elucidation of the phosphopeptide sequence. A higher number of sequence informative b and y ions were detected using a Q-TOF instead of an IT analyzer. In addition to the well-established phosphorylation sites at Ser(149) and Ser(127), this study also revealed the presence of two minor phosphorylation sites at Thr(145) and Ser(166). These findings indicate that RPLC-ESI-MS(2) on a Q-TOF analyzer is a useful technique for identifying low-abundance phosphorylation sites in caseins.