Publications by authors named "Paolo Donato"

8 Publications

  • Page 1 of 1

Manufacture of a Multi-Purpose Low-Cost Animal Bench-Model for Teaching Tracheostomy.

J Vis Exp 2019 05 18(147). Epub 2019 May 18.

Physiology and Experimental Surgery, Department of Translational Medicine, Università del Piemonte Orientale.

Tracheostomy is one of the most frequent procedures, performed through various techniques in the intensive care unit and emergency situations. Despite this, there is a lack of training on this procedure that affects its outcome, which is also dependent on operator's dexterity. Here, we take the specific training and simulation into consideration. This article aims to describe every step of the manufacture of a new multi-purpose low-cost animal bench-model, with the support of video and images, and to obtain an opinion about the quality of this model by administering a questionnaire to professionals with experience in the procedures. Ten experts in the technique were enrolled. The model scored an average of 3.45/5 for its anatomical realism; 4.75/5 for its usefulness as a training tool for simulation courses and assessments. The time necessary to build the model was 15 minutes, and the cost amounted to 10€. The animal bench-model was considered a very useful simulator for tracheostomy training and assessments. Therefore, it could be used as a tool for medical courses and residencies.
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http://dx.doi.org/10.3791/59396DOI Listing
May 2019

Analysis of Staphylococcus aureus-Infected Mice Reveals Differential Temporal and Spatial Expression Patterns of .

Infect Immun 2017 10 20;85(10). Epub 2017 Sep 20.

GSK Vaccines Srl, Siena, Italy

is an opportunistic human pathogen and a major cause of invasive infections such as bacteremia, endocarditis, pneumonia, and wound infections. FhuD2 is a staphylococcal lipoprotein involved in the uptake of iron-hydroxymate and is under the control of the iron uptake regulator Fur. This protein is part of an investigational multicomponent vaccine formulation that has shown protective efficacy in several murine models of infection. Even though expression has been shown to be upregulated in murine kidneys infected with , it is not known whether the bacterium undergoes increased iron deprivation during prolonged infection. Furthermore, different infection niches might provide different environments and levels of iron availability, resulting in different expression patterns among organs of the same host. To address these questions, we characterized the expression of the gene and confirmed Fur-dependent regulation of its expression. We further investigated its expression in mice infected with a bioluminescent reporter strain of expressing the luciferase operon under the control of the promoter. The emission of bioluminescence in different organs was followed over a 7-day time course, and quantitative real-time PCR analysis of the RNA transcribed from the endogenous gene was performed. Using this approach, we were able to show that expression was induced during infection in all organs analyzed and that differences in expression were observed at different time points and in different infected organs. Our data suggest that undergoes increased iron deprivation during the progression of infection in diverse host organs and accordingly induces dedicated iron acquisition mechanisms. Since FhuD2 plays a central role in providing the pathogen with the required iron, further knowledge of the patterns of expression during infection will be instrumental in better defining the role of this antigen in pathogenesis and as a vaccine antigen.
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http://dx.doi.org/10.1128/IAI.00270-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5607405PMC
October 2017

Staphylococcus aureus-dependent septic arthritis in murine knee joints: local immune response and beneficial effects of vaccination.

Sci Rep 2016 11 30;6:38043. Epub 2016 Nov 30.

GSK Vaccines, Via Fiorentina 1, Siena, 53100, Italy.

Staphylococcus aureus is the major cause of human septic arthritis and osteomyelitis, which deserve special attention due to their rapid evolution and resistance to treatment. The progression of the disease depends on both bacterial presence in situ and uncontrolled disruptive immune response, which is responsible for chronic disease. Articular and bone infections are often the result of blood bacteremia, with the knees and hips being the most frequently infected joints showing the worst clinical outcome. We report the development of a hematogenous model of septic arthritis in murine knees, which progresses from an acute to a chronic phase, similarly to what occurs in humans. Characterization of the local and systemic inflammatory and immune responses following bacterial infection brought to light specific signatures of disease. Immunization of mice with the vaccine formulation we have recently described (4C-Staph), induced a strong antibody response and specific CD4+ effector memory T cells, and resulted in reduced bacterial load in the knee joints, a milder general inflammatory state and protection against bacterial-mediated cellular toxicity. Possible correlates of protection are finally proposed, which might contribute to the development of an effective vaccine for human use.
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http://dx.doi.org/10.1038/srep38043DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5128924PMC
November 2016

Perioperative and periprocedural airway management and respiratory safety for the obese patient: 2016 SIAARTI Consensus.

Minerva Anestesiol 2016 12 19;82(12):1314-1335. Epub 2016 Oct 19.

Anesthesia and Intensive Care Unit, AOU Policlinico Vittorio Emanuele University Hospital, Catania, Italy.

Proper management of obese patients requires a team vision and appropriate behaviors by all health care providers in hospital. Specialist competencies are fundamental, as are specific clinical pathways and good clinical practices designed to deal with patients whose Body Mass Index (BMI) is ≥30 kg/m2. Standards of care for bariatric and non-bariatric surgery and for the critical care management of this population exist but are not well defined nor clearly followed in every hospital. Thus every anesthesiologist is likely to deal with this challenging population. Obesity is a multisystem, chronic, proinflammatory disorder. Unfortunately many countries are facing a marked increase in the obese population, defined as "globesity". Obesity presents an added risk in hospital, leading health care organizations to call for action to avoid adverse events and preventable complications. Periprocedural assessment and critical care strategies designed specifically for obese patients are crucial for reducing morbidity and mortality during surgery and in emergency settings, critical care and other particular settings (e.g., obstetrics). Specific care is needed for airway management, as are proactive strategies to reduce the risk of cardiovascular, endocrine, metabolic and infective complications; any effort can be fruitful, including special attention to the science of human factors. The Italian Society of Anesthesia, Analgesia, Resuscitation and Intensive Care (SIAARTI) organized a consensus project involving other national scientific societies to increase risk awareness, define the best multidisciplinary approach for treating obese patients in election and emergency, and enable every hospital to provide appropriate levels of care and good clinical practices. The Obesity Project Task Force, a section of the SIAARTI Airway Management Study Group, used a formal consensus process to identify a series of notes, alerts and statements, to be adopted as bundles, to define appropriate clinical pathways for hospitalized obese patients. The consensus, approved by the Task Force and endorsed by several European scientific societies actively operating in this field, is presented herein.
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December 2016

SslE elicits functional antibodies that impair in vitro mucinase activity and in vivo colonization by both intestinal and extraintestinal Escherichia coli strains.

PLoS Pathog 2014 May 8;10(5):e1004124. Epub 2014 May 8.

Novartis Vaccines and Diagnostics Srl, Siena, Italy.

SslE, the Secreted and surface-associated lipoprotein from Escherichia coli, has recently been associated to the M60-like extracellular zinc-metalloprotease sub-family which is implicated in glycan recognition and processing. SslE can be divided into two main variants and we recently proposed it as a potential vaccine candidate. By applying a number of in vitro bioassays and comparing wild type, knockout mutant and complemented strains, we have now demonstrated that SslE specifically contributes to degradation of mucin substrates, typically present in the intestine and bladder. Mutation of the zinc metallopeptidase motif of SslE dramatically impaired E. coli mucinase activity, confirming the specificity of the phenotype observed. Moreover, antibodies raised against variant I SslE, cloned from strain IHE3034 (SslEIHE3034), are able to inhibit translocation of E. coli strains expressing different variants through a mucin-based matrix, suggesting that SslE induces cross-reactive functional antibodies that affect the metallopeptidase activity. To test this hypothesis, we used well-established animal models and demonstrated that immunization with SslEIHE3034 significantly reduced gut, kidney and spleen colonization by strains producing variant II SslE and belonging to different pathotypes. Taken together, these data strongly support the importance of SslE in E. coli colonization of mucosal surfaces and reinforce the use of this antigen as a component of a broadly protective vaccine against pathogenic E. coli species.
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http://dx.doi.org/10.1371/journal.ppat.1004124DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4014459PMC
May 2014

Protective activity of Streptococcus pneumoniae Spr1875 protein fragments identified using a phage displayed genomic library.

PLoS One 2012 3;7(5):e36588. Epub 2012 May 3.

The Elie Metchnikoff Department, Università di Messina, Messina, Italy.

There is considerable interest in pneumococcal protein antigens capable of inducing serotype-independent immunoprotection and of improving, thereby, existing vaccines. We report here on the immunogenic properties of a novel surface antigen encoded by ORF spr1875 in the R6 strain genome. An antigenic fragment encoded by spr1875, designated R4, was identified using a Streptococcus pneumoniae phage displayed genomic library after selection with a human convalescent serum. Immunofluorescence analysis with anti-R4 antisera showed that Spr1875 was expressed on the surface of strains belonging to different serotypes. Moreover, the gene was present with little sequence variability in 27 different pneumococcal strains isolated worldwide. A mutant lacking Spr1875 was considerably less virulent than the wild type D39 strain in an intravenous mouse model of infection. Moreover, immunization with the R4 recombinant fragment, but not with the whole Spr1875 protein, induced significant protection against sepsis in mice. Lack of protection after immunization with the whole protein was related to the presence of immunodominant, non-protective epitopes located outside of the R4 fragment. In conclusion, our data indicate that Spr1875 has a role in pneumococcal virulence and is immunogenic. As the R4 fragment conferred immunoprotection from experimental sepsis, selected antigenic fragments of Spr1875 may be useful for the development of a pneumococcal protein-based vaccine.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0036588PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3343019PMC
January 2013

Multi high-throughput approach for highly selective identification of vaccine candidates: the Group A Streptococcus case.

Mol Cell Proteomics 2012 Jun 27;11(6):M111.015693. Epub 2012 Jan 27.

Novartis Vaccines and Diagnostics, Via Fiorentina 1, 53100 Siena, Italy.

We propose an experimental strategy for highly accurate selection of candidates for bacterial vaccines without using in vitro and/or in vivo protection assays. Starting from the observation that efficacious vaccines are constituted by conserved, surface-associated and/or secreted components, the strategy contemplates the parallel application of three high throughput technologies, i.e. mass spectrometry-based proteomics, protein array, and flow-cytometry analysis, to identify this category of proteins, and is based on the assumption that the antigens identified by all three technologies are the protective ones. When we tested this strategy for Group A Streptococcus, we selected a total of 40 proteins, of which only six identified by all three approaches. When the 40 proteins were tested in a mouse model, only six were found to be protective and five of these belonged to the group of antigens in common to the three technologies. Finally, a combination of three protective antigens conferred broad protection against a panel of four different Group A Streptococcus strains. This approach may find general application as an accelerated and highly accurate path to bacterial vaccine discovery.
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http://dx.doi.org/10.1074/mcp.M111.015693DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3433891PMC
June 2012

Peptide mimics of the group B meningococcal capsule induce bactericidal and protective antibodies after immunization.

J Immunol 2007 Apr;178(7):4417-23

Dipartimento di Scienze Microbiologiche, Genetiche e Molecolari, Università degli Studi di Messina, Messina, Italy.

Neisseria meningitidis serogroup B (MenB) is a leading cause of sepsis and meningitis in children. No vaccine is available for the prevention of these infections because the group B capsular polysaccharide (CP) (MenB CP) is unable to stimulate an immune response, due to its similarity with human polysialic acid. Because the MenB CP bears both human cross-reactive and non-cross-reactive determinants, we developed immunogenic peptide mimics of the latter epitopes. Peptides were selected from phage display libraries for their ability to bind to a protective anti-MenB CP mAb. One of these peptides (designated 9M) induced marked elevations in serum bactericidal activity, but not polysialic acid cross-reacting Abs, after gene priming followed by carrier-conjugate boosting. Moreover, the occurrence of bacteremia was prevented in infant rats by administration of immune sera before MenB challenge. 9M is a promising lead candidate for the development of an effective and affordable anti-MenB vaccine.
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http://dx.doi.org/10.4049/jimmunol.178.7.4417DOI Listing
April 2007