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Introduction: Procalcitonin is a marker for bacterial diseases and has been used to guide antibiotic prescription. Procalcitonin accuracy, measured at admission, in patients with community-acquired pneumonia (CAP), is unknown in the current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Objectives: To evaluate the diagnostic accuracy of procalcitonin to assess the need for antibiotic treatment in patients with CAP presenting to the emergency department during the SARS-CoV-2 pandemic. Methods: We performed a real-world diagnostic retrospective accuracy study of procalcitonin in patients admitted to the emergency department. Measures of diagnostic accuracy were calculated based on procalcitonin results compared to the reference standard of combined microbiological and radiological analysis. Sensitivity, specificity, positive and negative predictive values, and area under (AUC) the receiver-operating characteristic (ROC) curve were calculated in two analyses: first assessing procalcitonin ability to differentiate microbiologically proven bacteria from viral CAP and then clinically diagnosed bacterial CAP from viral CAP. Results: When using a procalcitonin threshold of 0.5 ng/mL to identify bacterial etiology within patients with CAP, we observed sensitivity and specificity of 50% and 64.1%, and 43% and 82.6%, respectively, in the two analyses. The positive and negative predictive values of a procalcitonin threshold of 0.5 ng/mL to identify patients for whom antibiotics should be advised were 46.4% and 79.7%, and 48.9% and 79% in the two analyses, respectively. The AUC for the two analyses was 0.60 (95% confidence interval [CI] 0.52–0.68) and 0.62 (95% CI, 0.55–0.69). Conclusions: Procalcitonin measured upon admission during the SARS-CoV-2 pandemic should not guide antibiotic treatment in patients with CAP.

Pseudomonas tolaasii, the causal organism of brown blotch disease of Agaricus bisporus and of the yellowing of Pleurotus ostreatus, was shown to produce in culture tolaasin I (1), tolaasin II (2), and five other minor metabolites, tolaasins A, B, C, D, and E (3-7). These compounds were demonstrated to be important in the development of the disease symptoms. This paper reports on the structural elucidation, based essentially on NMR studies and MS spectra, and biological activity of the above lipodepsipeptides (3-7). All the above analogues showed differences in the peptide moiety, as observed in other lipodepsipeptides of bacterial origin, and maintained the beta-hydroxyoctanoyl phi chain at the N-terminus, except tolaasin A, in which the acyl moiety was a gamma-carboxybutanoyl phi moiety. Among the target microorganisms used (fungi, yeast, and bacteria) the Gram-positive bacteria were the most sensitive, although the antimicrobial activity appeared to be correlated to the structural modification in the different analogues. The structure-activity relationships of these toxins are discussed.

The role of the exopolysaccharides (EPSs) produced by plant pathogenic bacteria has not completely clarified, they are considered either molecules able to avoid or delay the activation of plant defences, or acting as signal in the plant-pathogen cross-talk. In order to understand whether EPSs are recognized by infected plant cells and are able to induce the activation of plant defence responses, their capability to induce metabolic alteration in tobacco cells has been analysed. The results indicate that several EPSs, even if not chemically related, induce increases in phenylalanine ammonia-lyase, a marker enzyme of defence responses of plants against stress; but others are completely ineffective. The EPSs affecting phenylalanine ammonia-lyase also induce an increase in hydrogen peroxide production. Moreover, they alter the metabolism of ascorbate, another parameter indicative of the presence of stress conditions and the involvement of which in the hypersensitive reaction has been recently reported. The possibility that specific EPSs could act as signals in the plant-pathogenic bacteria interaction is discussed.

The biological activities of the lipodepsipeptides (LDP) white line–inducing principle (WLIP), produced by Pseudomonas reactans NCPPB1311, and tolaasin I, produced by P. tolaasii NCPPB2192, were compared. Antimicrobial assays showed that both LDP inhibited the growth of fungi—including the cultivated mushrooms Agaricus bisporus, Lentinus edodes, and Pleurotus spp.—chromista, and gram-positive bacteria. Assays of the two LDP on blocks of Agaricus bisporus showed their capacity to alter the mushrooms’ pseudo-tissues though WLIP was less active than that of tolaasin I. Contrary to previous studies, tolaasin I was found to inhibit the growth of gram-negative bacteria belonging to the genera Escherichia, Erwinia, Agrobacterium, Pseudomonas, and Xanthomonas. The only gram-negative bacterium affected by WLIP was Erwinia carotovora subsp. carotovora. Both WLIP and tolaasin I caused red blood cell lysis through a colloid-osmotic shock mediated by transmembrane pores; however, the haemolytic activity of WLIP was greater than that of tolaasin I. Transmembrane pores, at a concentration corresponding to 1.5 × C50, showed a radius between 1.5 and 1.7 ± 0.1 nm for WLIP and 2.1 ± 0.1 nm for tolaasin I. The antifungal activity of WLIP together with the finding that avirulent morphological variants of P. reactans lack WLIP production suggests that WLIP may play an important role in the interaction of the producing bacterium P. reactans and cultivated mushrooms. Additional keywords: Pseudomonas tolaasii.

The activity of tolaasin I and White Line Inducing Principle (WLIP), produced in vitro by virulent strains of Pseudomonas tolaasii and P. “reactans”, respectively, was comparatively evaluated on red blood cells and large unilamellar vesicles. Tolaasin I and WLIP caused red blood cell haemolysis through a colloid-osmotic shock apparently caused by transmembrane pores formation. Both lipodepsipeptides were able to form channels in lipid vesicles. Their activity was dependent on the toxin concentration and liposomes composition, and independent on the pH. Studies of dynamic light scattering suggested a detergent-like activity for WLIP at high concentration (higher than 27 µM). This effect was not detected for tolaasin I at the concentrations tested (up to 28.2 µM).

The 18-amino acid cytolytic lipodepsipeptide tolaasin, produced in culture by virulent strains of Pseudomonas tolaasii, is the causal agent of the brown blotch disease of the cultivated mushroom. Tolaasin has a sequence of D-amino acids in its N-terminal region, then alternates L- and D-amino acids, and bears a C-terminal lactone macrocycle composed of 5-residues. The solution structure of tolaasin in sodium dodecyl sulfate was studied by 2D-NMR spectroscopy and molecular dynamics simulated annealing calculations. Tolaasin forms an amphipathic left-handed alpha-helix in the regionDPro2-DalloThr14 comprising the sequence of seven D-amino acids and the adjacent L-D-L-D-D-region. To the best of our knowledge, this is the first recognized example of a left-handed alpha-helix including both D- and L-amino acids. The lactone macrocycle adopts a "boat-like" conformation and is shifted from the helical axis as to form a "golf-club" overall conformation. These structural features will be of importance in understanding, and preventing, tolaasin's role in the bacterial colonization of the host plant, and its toxic action on cells. Furthermore, the observed antimicrobial activity together with the potential resistance to enzymatic degradation and the increased antigenicity (both due to the presence of L- and D-amino acids) strongly suggests for tolaasin a potential role as a template model for the design of new therapeutic antibacterial molecules.