Collective behavior and virulence arsenal of the fish pathogen Piscirickettsia salmonis in the biofilm realm.

Piscirickettsiosis is a fish disease caused by the Gram-negative bacterium Piscirickettsia salmonis. This disease has a high socio-economic impact on the Chilean salmonid aquaculture industry. The bacterium has a cryptic character in the environment and their main reservoirs are yet unknown. Bacterial biofilms represent a ubiquitous mechanism of cell persistence in diverse natural environments and a risk factor for the pathogenesis of several infectious diseases, but their microbiological significance for waterborne veterinary diseases, including piscirickettsiosis, have seldom been evaluated. This study analyzed the in vitro biofilm behavior of P. salmonis LF-89T (genogroup LF-89) and CA5 (genogroup EM-90) using a multi-method approach and elucidated the potential arsenal of virulence of the P. salmonis LF-89T type strain in its biofilm state. P. salmonis exhibited a quick kinetics of biofilm formation that followed a multi-step and highly strain-dependent process. There were no major differences in enzymatic profiles or significant differences in cytotoxicity (as tested on the Chinook salmon embryo cell line) between biofilm-derived bacteria and planktonic equivalents. The potential arsenal of virulence of P. salmonis LF-89T in biofilms, as determined by whole-transcriptome sequencing and differential gene expression analysis, consisted of genes involved in cell adhesion, polysaccharide biosynthesis, transcriptional regulation, and gene mobility, among others. Importantly, the global gene expression profiles of P. salmonis LF-89T were not enriched with virulence-related genes upregulated in biofilm development stages at 24 and 48 h. An enrichment in virulence-related genes exclusively expressed in biofilms was also undetected. These results indicate that early and mature biofilm development stages of P. salmonis LF-89T were transcriptionally no more virulent than their planktonic counterparts, which was supported by cytotoxic trials, which, in turn, revealed that both modes of growth induced important and very similar levels of cytotoxicity on the salmon cell line. Our results suggest that the aforementioned biofilm development stages do not represent hot spots of virulence compared with planktonic counterparts. This study provides the first transcriptomic catalogue to select specific genes that could be useful to prevent or control the (in vitro and/or in vivo) adherence and/or biofilm formation by P. salmonis and gain further insights into piscirickettsiosis pathogenesis.

Butyric acid stimulates bovine neutrophil functions and potentiates the effect of platelet activating factor.

Increased short-chain fatty acid (SCFA) production is associated with subacute ruminal acidosis (SARA) and activation of inflammatory processes. In humans and rodents, SCFAs modulate inflammatory responses in the gut via free fatty acid receptor 2 (FFA2). In bovines, butyric acid is one of the most potent FFA2 agonists. Its expression in bovine neutrophils has recently been demonstrated, suggesting a role in innate immune response in cattle. This study aimed to evaluate if butyric acid modulates oxidative and non-oxidative functions or if it can potentiate other inflammatory mediators in bovine neutrophils. Our results showed that butyric acid can activate bovine neutrophils, inducing calcium (Ca(2+)) influx and mitogen-activated protein kinase (MAPK) phosphorylation, two second messengers involved in FFA2 activation. Ca(2+) influx induced by butyric acid was dependent on the extracellular and intracellular Ca(2+) source and phospholipase C (PLC) activation. Butyric acid alone had no significant effect on reactive oxygen species (ROS) production and chemotaxis; however, a priming effect on platelet-activating factor (PAF), a potent inflammatory mediator, was observed. Butyric acid increased CD63 expression and induced the release of neutrophil granule markers matrix metalloproteinase-9 (MMP-9) and lactoferrin. Finally, we observed that butyric acid induced neutrophil extracellular trap (NET) formation without affecting cellular viability. These findings suggest that butyric acid, a component of the ruminal fermentative process, can modulate the innate immune response of ruminants.

ESR, electrochemical and cyclodextrin-inclusion studies of triazolopyridyl pyridyl ketones and dipyridyl ketones derivatives.

The electron spin resonance (ESR) spectra of free radicals obtained by electrolytic reduction of triazolopyridyl pyridyl ketones and dipyridyl ketones derivatives were measured in dimethylsulfoxide (DMSO). The hyperfine patterns indicate that the spin density delocalization is dependent of the rings presented in the molecule. The electrochemistry of these compounds was characterized using cyclic voltammetry, in DMSO as solvent. When one carbonyl is present in the molecule one step in the reduction mechanism was observed while two carbonyl are present two steps were detected. The first wave was assigned to the generation of the correspondent free radical species, and the second wave was assigned to the dianion derivatives. The phase-solubility measurements indicated an interaction between molecules selected and cyclodextrins in water. These inclusion complexes are 1:1 with betaCD, and HP-betaCD. The values of Ks showed a different kind of complexes depending on which rings are included. AM1 and DFT calculations were performed to obtain the optimized geometries, theoretical hyperfine constants, and spin distributions, respectively. The theoretical results are in complete agreement with the experimental ones.

ESR and electrochemical studies of 2-acylpyridines and 6,6'-diacyl-2,2'-bipyridines.

The ESR spectra of radicals obtained by electrolytic reduction of 2-acylpyridines and 6,6'-diacyl-2,2'-bipyridines were measured in dimethylsulfoxide (DMSO) and analyzed by quantum chemical calculations. The electrochemistry of these compounds was characterized using cyclic voltammetry, in DMSO solvent. The results showed a two step reduction mechanism, first wave was assigned to the generation of the correspondent free radical species, and the second wave was assigned to the dianion derivatives. AM1 and DFT calculations were performed to obtain the optimized geometries, theoretical hyperfine constants, and spin distributions, respectively. The theoretical results are in complete agreement with the experimental ones.

Práctica contemporánea de la angioplastia coronaria en Chile: informe final del Registro Nacional de Angioplastia Coronaria (RENAC) 2001-2002 / Final report of the Chilean National Coronary Angioplasty Registry (RENAC)

Background:Coronary angioplasty was introduced in Chile in 1982, but only after the introduction of stents it became commonplace. However, the results of this procedure at the national level remain largely unknown. Aim: To assess the results and characteristics of coronary angioplasty and develop a national registry in Chile (RENAC). Patients and Methods: All centers performing angioplasty were invited to contribute prospectively with the clinical, angiographic, procedural characteristics and results of all consecutive coronary angioplasty procedures attempted between June 2001 and October 2002. Results: In 10 centers, 1768 lesions were treated in 1484 procedures (22.98 percent in women). Mean age was 60.8±11.3 years old. Diabetes was found in 21.8 percent, hypertension in 60.2 percent, dyslipidemia in 52.0 percent, active smoking in 40.0 percent, renal failure in 6.2 percent. Myocardial infarction was recent in 28.4 percent and acute in 19.7 percent. Forty eight percent of patients had unstable angina and 15.5 percent had stable angina. Fifty three percent of patients had single vessel disease and 71 percent, normal left ventricular eyection fraction. One vessel was treated in 90.6 percent of patients and 81.7 percent of lesions were treated with stents, 17.9 percent only with baloon and in 0.4 percent with atherectomy. Angiographic success was obtained in 95.2 percent. Clinical success was obtained in 92.2 percent, and 95.1 percent in patients without acute myocardial infarction. Overall inhospital death was 2.2 percent. In patients without myocardial infarction, the figure was 1.1 percent. Conclusions: Coronary angioplasty in Chile is performed mostly for the treatment of acute coronary syndromes. Stents are the most frequently used devices. The high success, low mortality and complications observed are comparable to North American registries (Rev Méd Chile 2004; 132: 913-22).