Publications by authors named "Luisa Lanzilao"

11 Publications

  • Page 1 of 1

Improvement in estimation of time since death by albumin and potassium concentrations in vitreous humor.

Forensic Sci Int 2020 Sep 27;314:110393. Epub 2020 Jun 27.

Section of Forensic Medical Sciences, Department of Health Sciences, University of Florence, Florence, Italy.

The measurement of the potassium concentration (K) in vitreous humor (VH) has been a well-known adjunct for the estimation of the post-mortem interval (PMI) since the early 1960s. For years, however, many authors have been using other biochemical markers in an attempt to improve predictions. In this paper we confirm the role of K in the determination of the PMI adopting a linear regression model and we investigate whether other biochemical markers could improve the model through a multiple regression analysis. Additionally, the research aims to confirm the data of the analytes of interest among different techniques and instrumentations. We deemed this as an important issue because a primary concern in the literature is that automated analytical methods are often calibrated and, for the most part, validated for serum or urine analysis. Our results confirmed the well-established role of K as well as highlighted Albumin as a novel marker to be considered for further improvement of prediction models, especially since 72 h after death.
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http://dx.doi.org/10.1016/j.forsciint.2020.110393DOI Listing
September 2020

Prognostic role of intrathecal IgM synthesis in multiple sclerosis: Results from a clinical series.

Mult Scler 2021 Feb 24;27(2):198-207. Epub 2020 Feb 24.

Department of Neurofarba, University of Florence, Florence, Italy/IRCCS Fondazione Don Carlo Gnocchi, Florence, Italy.

Background: There is emerging evidence that intrathecal IgM synthesis (ITMS) is a risk factor for conversion to clinically defined multiple sclerosis (CDMS) in clinically isolated syndrome (CIS) patients.

Objectives: The objective of this study is to verify the prognostic role of ITMS as a risk factor for the second clinical attack in patients after the first demyelinating event.

Methods: Monocentric observational study performed on prospectively acquired clinical data and retrospective evaluation of magnetic resonance imaging (MRI) data. ITMS was assessed according to Reiber's non-linear function. We compared time to the second attack by using Kaplan-Meier curves and performed adjustment by Cox regression analysis.

Results: Demographics and clinical data were collected prospectively in a cohort of 68 patients. ITMS occurred in 40% (27/68) of patients who had a higher T1-hypointense lesion load at brain MRI ( = 0.041). In multivariate Cox regression analysis (adjusted for age, sex, baseline Expanded Disability Status Scale, IgG oligoclonal bands and disease-modifying treatment exposure), relapsing-remitting multiple sclerosis (MS) patients with ITMS were at higher risk to experience a second clinical attack (adjusted hazard ratio (aHR) = 6.3, 95% confidence interval (CI) = 2.1-18.4,  = 0.001).

Conclusion: Together with previous studies, our findings support the role of ITMS as a prognostic biomarker in MS.
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http://dx.doi.org/10.1177/1352458520907913DOI Listing
February 2021

Comparative immunogenicity and efficacy of equivalent outer membrane vesicle and glycoconjugate vaccines against nontyphoidal .

Proc Natl Acad Sci U S A 2018 10 27;115(41):10428-10433. Epub 2018 Sep 27.

Jenner Institute, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7DQ, United Kingdom.

Nontyphoidal cause a devastating burden of invasive disease in sub-Saharan Africa with high levels of antimicrobial resistance. Vaccination has potential for a major global health impact, but no licensed vaccine is available. The lack of commercial incentive makes simple, affordable technologies the preferred route for vaccine development. Here we compare equivalent Generalized Modules for Membrane Antigens (GMMA) outer membrane vesicles and O-antigen-CRM glycoconjugates to deliver lipopolysaccharide O-antigen in bivalent Typhimurium and Enteritidis vaccines. strains were chosen and deleted to induce GMMA production. O-antigens were extracted from wild-type bacteria and conjugated to CRM Purified GMMA and glycoconjugates were characterized and tested in mice for immunogenicity and ability to reduce infection. GMMA and glycoconjugate O-antigen had similar structural characteristics, O-acetylation, and glucosylation levels. Immunization with GMMA induced higher anti-O-antigen IgG than glycoconjugate administered without Alhydrogel adjuvant. With Alhydrogel, antibody levels were similar. GMMA induced a diverse antibody isotype profile with greater serum bactericidal activity than glycoconjugate, which induced almost exclusively IgG1. Immunization reduced bacterial colonization of mice subsequently infected with Typhimurium numbers were lower in tissues of mice vaccinated with GMMA compared with glycoconjugate. Enteritidis burden in the tissues was similar in mice immunized with either vaccine. With favorable immunogenicity, low cost, and ability to induce functional antibodies and reduce bacterial burden, GMMA offer a promising strategy for the development of a nontyphoidal vaccine compared with established glycoconjugates. GMMA technology is potentially attractive for development of vaccines against other bacteria of global health significance.
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http://dx.doi.org/10.1073/pnas.1807655115DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6187145PMC
October 2018

Immunogenicity of a Bivalent Adjuvanted Glycoconjugate Vaccine against Typhimurium and Enteritidis.

Front Immunol 2017 27;8:168. Epub 2017 Feb 27.

Laboratorio di Microbiologia Molecolare e Biotecnologia (LA.M.M.B.), Dipartimento di Biotecnologie Mediche, Università di Siena , Siena , Italy.

serovars Typhimurium and Enteritidis are the predominant causes of invasive non-typhoidal (iNTS) disease. Considering the co-endemicity of . Typhimurium and . Enteritidis, a bivalent vaccine formulation against both pathogens is necessary for protection against iNTS disease, thus investigation of glycoconjugate combination is required. In the present work, we investigated the immune responses induced by . Typhimurium and . Enteritidis monovalent and bivalent glycoconjugate vaccines adjuvanted with aluminum hydroxide (alum) only or in combination with cytosine-phosphorothioate-guanine oligodeoxynucleotide (CpG). Humoral and cellular, systemic and local, immune responses were characterized in two different mouse strains. All conjugate vaccines elicited high levels of serum IgG against the respective O-antigens (OAg) with bactericidal activity. The bivalent conjugate vaccine induced systemic production of antibodies against both . Typhimurium and . Enteritidis OAg. The presence of alum or alum + CpG adjuvants in vaccine formulations significantly increased the serum antigen-specific antibody production. The alum + CpG bivalent vaccine formulation triggered the highest systemic anti-OAg antibodies and also a significant increase of anti-OAg IgG in intestinal washes and fecal samples, with a positive correlation with serum levels. These data demonstrate the ability of monovalent and bivalent conjugate vaccines against . Typhimurium and . Enteritidis to induce systemic and local immune responses in different mouse strains, and highlight the suitability of a bivalent glycoconjugate formulation, especially when adjuvanted with alum + CpG, as a promising candidate vaccine against iNTS disease.
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http://dx.doi.org/10.3389/fimmu.2017.00168DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5326758PMC
February 2017

Strain Selection for Generation of O-Antigen-Based Glycoconjugate Vaccines against Invasive Nontyphoidal Salmonella Disease.

PLoS One 2015 7;10(10):e0139847. Epub 2015 Oct 7.

Sclavo Behring Vaccines Institute for Global Health S.r.l., a GSK company (formerly Novartis Vaccines Institute for Global Health S.r.l), Via Fiorentina 1, 53100, Siena, Italy.

Nontyphoidal Salmonellae, principally S. Typhimurium and S. Enteritidis, are a major cause of invasive bloodstream infections in sub-Saharan Africa with no vaccine currently available. Conjugation of lipopolysaccharide O-antigen to a carrier protein constitutes a promising vaccination strategy. Here we describe a rational process to select the most appropriate isolates of Salmonella as source of O-antigen for developing a bivalent glycoconjugate vaccine. We screened a library of 30 S. Typhimurium and 21 S. Enteritidis in order to identify the most suitable strains for large scale O-antigen production and generation of conjugate vaccines. Initial screening was based on growth characteristics, safety profile of the isolates, O-antigen production, and O-antigen characteristics in terms of molecular size, O-acetylation and glucosylation level and position, as determined by phenol sulfuric assay, NMR, HPLC-SEC and HPAEC-PAD. Three animal isolates for each serovar were identified and used to synthesize candidate glycoconjugate vaccines, using CRM197 as carrier protein. The immunogenicity of these conjugates and the functional activity of the induced antibodies was investigated by ELISA, serum bactericidal assay and flow cytometry. S. Typhimurium O-antigen showed high structural diversity, including O-acetylation of rhamnose in a Malawian invasive strain generating a specific immunodominant epitope. S. Typhimurium conjugates provoked an anti-O-antigen response primarily against the O:5 determinant. O-antigen from S. Enteritidis was structurally more homogeneous than from S. Typhimurium, and no idiosyncratic antibody responses were detected for the S. Enteritidis conjugates. Of the three initially selected isolates, two S. Typhimurium (1418 and 2189) and two S. Enteritidis (502 and 618) strains generated glycoconjugates able to induce high specific antibody levels with high breadth of serovar-specific strain coverage, and were selected for use in vaccine production. The strain selection approach described is potentially applicable to the development of glycoconjugate vaccines against other bacterial pathogens.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0139847PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4596569PMC
June 2016

Relationship between antibody susceptibility and lipopolysaccharide O-antigen characteristics of invasive and gastrointestinal nontyphoidal Salmonellae isolates from Kenya.

PLoS Negl Trop Dis 2015 Mar 4;9(3):e0003573. Epub 2015 Mar 4.

Novartis Vaccines Institute for Global Health (NVGH), Siena, Italy.

Background: Nontyphoidal Salmonellae (NTS) cause a large burden of invasive and gastrointestinal disease among young children in sub-Saharan Africa. No vaccine is currently available. Previous reports indicate the importance of the O-antigen of Salmonella lipopolysaccharide for virulence and resistance to antibody-mediated killing. We hypothesised that isolates with more O-antigen have increased resistance to antibody-mediated killing and are more likely to be invasive than gastrointestinal.

Methodology/principal Findings: We studied 192 NTS isolates (114 Typhimurium, 78 Enteritidis) from blood and stools, mostly from paediatric admissions in Kenya 2000-2011. Isolates were tested for susceptibility to antibody-mediated killing, using whole adult serum. O-antigen structural characteristics, including O-acetylation and glucosylation, were investigated. Overall, isolates were susceptible to antibody-mediated killing, but S. Enteritidis were less susceptible and expressed more O-antigen than Typhimurium (p<0.0001 for both comparisons). For S. Typhimurium, but not Enteritidis, O-antigen expression correlated with reduced sensitivity to killing (r = 0.29, 95% CI = 0.10-0.45, p = 0.002). Both serovars expressed O-antigen populations ranging 21-33 kDa average molecular weight. O-antigen from most Typhimurium were O-acetylated on rhamnose and abequose residues, while Enteritidis O-antigen had low or no O-acetylation. Both Typhimurium and Enteritidis O-antigen were approximately 20%-50% glucosylated. Amount of S. Typhimurium O-antigen and O-antigen glucosylation level were inversely related. There was no clear association between clinical presentation and antibody susceptibility, O-antigen level or other O-antigen features.

Conclusion/significance: Kenyan S. Typhimurium and Enteritidis clinical isolates are susceptible to antibody-mediated killing, with degree of susceptibility varying with level of O-antigen for S. Typhimurium. This supports the development of an antibody-inducing vaccine against NTS for Africa. No clear differences were found in the phenotype of isolates from blood and stool, suggesting that the same isolates can cause invasive disease and gastroenteritis. Genome studies are required to understand whether invasive and gastrointestinal isolates differ at the genotypic level.
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http://dx.doi.org/10.1371/journal.pntd.0003573DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4352093PMC
March 2015

Invasive African Salmonella Typhimurium induces bactericidal antibodies against O-antigens.

Microb Pathog 2013 Oct 10;63:19-23. Epub 2013 Jun 10.

Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.

Nontyphoidal Salmonella are a major and emerging cause of fatal invasive disease in Africa, and are genetically distinct from those found elsewhere in the world. Understanding the targets of protective immunity to these African Salmonellae is key to vaccine development. We immunized mice and rabbits with heat-inactivated wild-type African invasive Salmonella Typhimurium D23580 and rough mutants lacking O-antigen. Wild-type Salmonella, unlike rough bacteria, induced a large bactericidal antibody response mainly against O-antigen. Bactericidal ability of anti-O-antigen antibodies was confirmed following purification by affinity chromatography. The current findings support the development of an O-antigen conjugate vaccine against invasive nontyphoidal Salmonellae for Africa.
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http://dx.doi.org/10.1016/j.micpath.2013.05.014DOI Listing
October 2013

Characterization of Citrobacter sp. line 328 as a source of Vi for a Vi-CRM(197) glycoconjugate vaccine against Salmonella Typhi.

J Infect Dev Ctries 2012 Nov 26;6(11):763-73. Epub 2012 Nov 26.

Novartis Vaccines Institute for Global Health.

Introduction: Salmonella enterica serovar Typhi is the causative agent of typhoid fever with over 22 million cases and over 200,000 deaths reported annually. A vaccine is much needed for use in young children and the Novartis Vaccines Institute for Global Health (NVGH) is developing a conjugate vaccine which targets S. Typhi Vi capsular polysaccharide.

Methodology: Here we describe a method suitable for industrial scale production of the Vi antigen based on expression by a Citrobacter line. We optimized the production of Vi by selecting a suitable Citrobacter strain (Citrobacter 328) that yields high and stable expression of Vi in chemically defined medium under industrial-scale fermentation conditions.

Results: Vi-CRM197 made using Vi from Citrobacter 328 elicited high anti-Vi antibody levels in mice and rabbits.

Conclusions: Citrobacter 328 is a suitable strain for production of Vi for conjugate anti-Typhi vaccines. Being a BSL-1 organism, which grows in defined medium and stably produces high yields of Vi, it offers excellent potential for safe production of inexpensive vaccines for populations at risk of typhoid fever.
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http://dx.doi.org/10.3855/jidc.2495DOI Listing
November 2012

O:2-CRM(197) conjugates against Salmonella Paratyphi A.

PLoS One 2012 7;7(11):e47039. Epub 2012 Nov 7.

Novartis Vaccines Institute for Global Health, Siena, Italy.

Enteric fevers remain a common and serious disease, affecting mainly children and adolescents in developing countries. Salmonella enterica serovar Typhi was believed to cause most enteric fever episodes, but several recent reports have shown an increasing incidence of S. Paratyphi A, encouraging the development of a bivalent vaccine to protect against both serovars, especially considering that at present there is no vaccine against S. Paratyphi A. The O-specific polysaccharide (O:2) of S. Paratyphi A is a protective antigen and clinical data have previously demonstrated the potential of using O:2 conjugate vaccines. Here we describe a new conjugation chemistry to link O:2 and the carrier protein CRM(197), using the terminus 3-deoxy-D-manno-octulosonic acid (KDO), thus leaving the O:2 chain unmodified. The new conjugates were tested in mice and compared with other O:2-antigen conjugates, synthesized adopting previously described methods that use CRM(197) as carrier protein. The newly developed conjugation chemistry yielded immunogenic conjugates with strong serum bactericidal activity against S. Paratyphi A.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0047039PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3492368PMC
April 2013

Evaluation of the immunogenicity and biological activity of the Citrobacter freundii Vi-CRM197 conjugate as a vaccine for Salmonella enterica serovar Typhi.

Clin Vaccine Immunol 2011 Mar 19;18(3):460-8. Epub 2011 Jan 19.

Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy.

Typhoid fever remains a major health problem in developing countries. Young children are at high risk, and a vaccine effective for this age group is urgently needed. Purified capsular polysaccharide from Salmonella enterica serovar Typhi (Vi) is licensed as a vaccine, providing 50 to 70% protection in individuals older than 5 years. However, this vaccine is ineffective in infants. Vi conjugated to a carrier protein (i.e., an exoprotein A mutant from Pseudomonas aeruginosa [rEPA]) is highly immunogenic, provides long-term protection, and shows more than 90% protective efficacy in children 2 to 5 years old. Here, we describe an alternative glycoconjugate vaccine for S. Typhi, Vi-CRM(197), where Vi was obtained from Citrobacter freundii WR7011 and CRM(197), the mutant diphtheria toxin protein, was used as the carrier. We investigated the optimization of growth conditions for Vi production from C. freundii WR7011 and the immunogenicity of Vi-CRM(197) conjugates in mice. The optimal saccharide/protein ratio of the glycoconjugates was identified for the best antibody production. We also demonstrated the ability of this new vaccine to protect mice against challenge with Vi-positive Salmonella enterica serovar Typhimurium.
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http://dx.doi.org/10.1128/CVI.00387-10DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3067394PMC
March 2011

Curcumin protects cardiac cells against ischemia-reperfusion injury: effects on oxidative stress, NF-kappaB, and JNK pathways.

Free Radic Biol Med 2008 Sep 20;45(6):839-46. Epub 2008 Jun 20.

Department of Biochemical Sciences University of Florence, 50134 Florence, Italy.

In this study we explored the effects of curcumin in cardiac cells subjected to a protocol simulating ischemia-reperfusion (IR). Curcumin (10 microM) was administered before ischemia (pretreatment) or at the moment of reperfusion (posttreatment) and its effects were compared to those produced by a reference antioxidant (Trolox) with an equal antioxidant capacity. IR cardiac cells showed clear signs of oxidative stress, impaired mitochondrial activity, and a marked development of both necrotic and apoptotic processes; at the same time, increases in NF-kappaB nuclear translocation and JNK phosphorylation were observed. Curcumin pretreatment was revealed to be the most effective in attenuating all these modifications and, in particular, in reducing the death of IR cells. This confirms that the protective effect of curcumin is not related simply to its antioxidant properties but involves other mechanisms, notably interactions in the NF-kappaB and JNK pathways. These findings suggest that curcumin administration, in particular before the hypoxic challenge, represents a promising approach to protecting cardiac cells against IR injury. In this scenario our results point out the importance of the chronology for the outcome of the treatment and provide a differential valuation of the degree of protection that curcumin can exert by its antioxidant activity or by other mechanisms.
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http://dx.doi.org/10.1016/j.freeradbiomed.2008.06.013DOI Listing
September 2008