Efficacy of ozonated water as a PS in photodynamic therapy: A tool for dental caries management? An in vitro study

Background: The most prevalent noncommunicable disease in the world is dental caries; and when it is not adequately treated, it is usually associated with tooth loss or severe dental lesions. In fact, expensive care or tooth extraction may be necessary due to the negative effects dental caries have on general health. This is due to its frequent pain and secondary bacterial infections. The aim of this study was to investigate the activity of ozonated water as such and in combination with appropriate light radiation so as to perform a photodynamic treatment (PDT) against the cariogenic bacterium Streptococcus mutans. Design and methods: This work has been performed in vitro by using an S. mutans strain mainly structured in a biofilm status, reproducing the natural condition of the tooth infection. The ozone was tested at three different concentrations by using a commercial device able to generate different O3 formulations in water. The PDT treatment requires an appropriate light wavelength, evaluated in this work through the UV-Vis adsorption spectrum of the ozonated water. Results: The obtained results suggested an effective and synergic property of O3 and light at 460–470nm against this microorganism. The most antibiofilm activity was observed using a concentration of ozone of 0.06mg/L alone as well as with PDT treatment. Conclusions: The results are encouraging for additional research and in vitro/in vivo fresh experimental investigations to perform an exhaustive antimicrobial treatment protocol against the S. mutans tooth infection

Mouthwash Based on Ozonated Olive Oil in Caries Prevention: A Preliminary In-Vitro Study

(1) Background: Ozone (O3) proved to oxidize organic and inorganic compounds, and its efficacy against bacteria, viruses and fungi plasma membranes was of interest. Ozone vehicle can be a gaseous form, ozonated water or ozonized oil. The aim of this in-vitro study was to evaluate the efficacy of ozonated olive oil against Streptococcus mutans. (2) Methods: Two different commercial mouthwashes were tested: Ialozon Blu (IB) (Gemavip, Cagliari, Italy), with ozonated olive oil, and Ialozon Rose (IR) (Gemavip, Cagliari, Italy), with ozonated olive oil, hyaluronic acid and vitamin E. All formulates were analyzed in a dilution range from 2- to 256-folds in saline solution, as to reproduce the salivary dilution. Streptococcus mutans CIP103220 strain was used for the antimicrobial susceptibility test, and the Kirby-Bauer inhibition method was performed to evaluate the Minimum Inhibitory (MIC), Minimum Bactericidal (MBC), and Minimum Biofilm Inhibitory Concentration (MBIC). (3) Results: Both formulates showed the same antimicrobial activity. MIC, MBC, and MBIC were observed for dilution factors of 1/32, 1/8 and 1/8, respectively. The mean value of inhibition zone diameter was 16.5 mm for IB, and 18 mm for IR. (4) Conclusions: The results suggested that ozonized olive oil formulates were able to inactivate Streptococcus mutans avoiding the salivary dilution effect in the oral cavity

Implementation of at-line capillary zone electrophoresis for fast and reliable determination of adenovirus concentrations in vaccine manufacturing

A CZE method was validated and implemented for fast and accurate in-process determination of adenovirus concentrations of downstream process samples obtained during manufacturing of adenovirus vector-based vaccines. An analytical-quality-by-design approach was embraced for method development, method implementation, and method maintenance. CZE provided separation of adenovirus particles from sample matrix components, such as cell debris, residual DNA and proteins. The intermediate precision of the virus particle concentration was 6.9% RSD and the relative bias was 2.3%. In comparison, the CZE method is intended to replace a quantitative polymerase chain reaction method which requires three replicates in three analytical runs to achieve an intermediate precision of 8.1% RSD. Given that, in addition, the time from sampling till reporting results of the CZE method was less than 2 h, whereas quantitative polymerase chain reaction requires 3 days, it follows that the CZE method enables faster processing times in downstream processing

Development and application of a MLST panel for the identification of informative polymorphisms in Leishmania infantum strains in the Mediterranean region

Background. Leishmaniasis is a zoonotic disease endemic in the Mediterranean region, where the causative agent of human and canine infection is Leishmania infantum. The spread of leishmaniasis is associated with population movements, ecology of phlebotomine vectors, and reservoir host. We used multilocus sequence typing (MLST) to explore the genetic variability of L. infantum strains in the Mediterranean region, including the borderline territory of Pantelleria island, and identify informative polymorphisms for rapid identification of genotypes through high-resolution melt (HRM)-based assays. Material and Methods. A customized sequencing panel targeting 14 housekeeping genes was designed and MLST analysis was performed using the Ion Torrent S5 on 9 L. infantum strains/isolates: 5 canine isolates (3 from Pantelleria Island and 2 from central Italy), and 4 human isolates/strains from Tunisia, France, central and southern Italy. MLST results and in silico analysis of sequences available in Genbank allowed to select two informative polymorphisms on ME and GPI genes (390T/G and 1834A/G, respectively) used to develop two HRM-based assays for fast screening of 28 clinical samples. Results. The MLST analysis identified a single L. infantum clonal complex regardless of the geographic origin or host (human or canine), except for the human isolate from central Italy that showed polymorphisms in 11 out of 14 housekeeping genes, and clustered independently in a molecular phylogenetic analysis. Successively, the screening through HRM-based assays of 28 clinical samples from central/south Italy and Pantelleria island allowed to identify 6 diploid sequence types (DSTs). Interestingly, the sequence type 390T/1834A was found only in Pantelleria island (prevalence 75%). Conclusion. This study represents a description of the genetic variability of L. infantum through a first approach based on MLST and then by HRM analysis on selected polymorphisms. The HRM assays could be used as fast and cheap tools for epidemiological surveillance of L. infantum

Evaluation of antibody response to BNT162b2 mRNA COVID-19 vaccine in patients affected by immune-mediated inflammatory diseases up to 5 months after vaccination

SARS-CoV-2 vaccination with mRNA product BNT162b2 elicited high immunogenicity in healthy subjects in trials. This study aims to better understand the factors that influence the humoral immune response to vaccination against SARS-CoV-2 in patients with immune-mediated inflammatory diseases (IMIDs). We enrolled patients and healthy healthcare workers control group (HCW) that underwent mRNA BNT162b2 vaccination and measured the serum IgG anti-S-RBD response at booster dose (T1), one month after booster dose (T2) and up to 5 months (T3). Demographic, disease-specific and vaccination data were recorded. Vaccination response of 551 participants naïve to SARS-CoV-2 infection were included in HCW and 102 in the IMID group, analyzing separately those on anti-CD20. At T2 all naïve HCW developed anti-S-RBD-IgG, while 94% of IMID responded (p < 0.001). IMID patients had a significantly different level of IgG than HCW at both T1 (p = 0.031), T2 (p < 0.001), while there was no significant difference at T3. There were no statistically significant differences according to the IMID type or to ongoing treatment with immunosuppressants, corticosteroids or biological drugs other than anti-CD20. The proportion and magnitude of response was significantly lower in IMID treated with anti-CD20 drugs. There was a correlation with age at T1 and at T2 but not at T3, stronger in patients than in HCW. Immune response close after BNT162b2 vaccination is reduced in patients with IMID, but there is no significant difference at 5 months. The measured reduction is related to age and the disease itself rather than treatments, with the exception of anti-CD20 drugs

Gut Microbiota Diversity and C-Reactive Protein Are Predictors of Disease Severity in COVID-19 Patients

Funding Information: We acknowledge support from the NOVA Medical School of Universidade NOVA de Lisboa, CINTESIS, and CHRC. Funding. This study was sponsored by the Funda??o para a Ci?ncia e a Tecnologia (FCT, project no. 268_596883842), BIOCODEX, and CINTESIS (reference UIDB/4255/2020). The funders had no role in study design, data collection, data analysis, data interpretation, or manuscript writing. Publisher Copyright: © Copyright © 2021 Moreira-Rosário, Marques, Pinheiro, Araújo, Ribeiro, Rocha, Mota, Pestana, Ribeiro, Pereira, de Sousa, Pereira-Leal, de Sousa, Morais, Teixeira, Rocha, Silvestre, Príncipe, Gatta, Amado, Santos, Maltez, Boquinhas, de Sousa, Germano, Sarmento, Granja, Póvoa, Faria and Calhau.The risk factors for coronavirus disease 2019 (COVID-19) severity are still poorly understood. Considering the pivotal role of the gut microbiota on host immune and inflammatory functions, we investigated the association between changes in the gut microbiota composition and the clinical severity of COVID-19. We conducted a multicenter cross-sectional study prospectively enrolling 115 COVID-19 patients categorized according to: (1) the WHO Clinical Progression Scale—mild, 19 (16.5%); moderate, 37 (32.2%); or severe, 59 (51.3%), and (2) the location of recovery from COVID-19—ambulatory, 14 (household isolation, 12.2%); hospitalized in ward, 40 (34.8%); or hospitalized in the intensive care unit, 61 (53.0%). Gut microbiota analysis was performed through 16S rRNA gene sequencing, and the data obtained were further related to the clinical parameters of COVID-19 patients. The risk factors for COVID-19 severity were identified by univariate and multivariable logistic regression models. In comparison to mild COVID-19 patients, the gut microbiota of moderate and severe patients have: (a) lower Firmicutes/Bacteroidetes ratio; (b) higher abundance of Proteobacteria; and (c) lower abundance of beneficial butyrate-producing bacteria such as the genera Roseburia and Lachnospira. Multivariable regression analysis showed that the Shannon diversity index [odds ratio (OR) = 2.85, 95% CI = 1.09–7.41, p = 0.032) and C-reactive protein (OR = 3.45, 95% CI = 1.33–8.91, p = 0.011) are risk factors for severe COVID-19 (a score of 6 or higher in the WHO Clinical Progression Scale). In conclusion, our results demonstrated that hospitalized patients with moderate and severe COVID-19 have microbial signatures of gut dysbiosis; for the first time, the gut microbiota diversity is pointed out as a prognostic biomarker of COVID-19 severity.publishersversionpublishe

Human Leukocyte Antigen Complex and Other Immunogenetic and Clinical Factors Influence Susceptibility or Protection to SARS-CoV-2 Infection and Severity of the Disease Course. The Sardinian Experience

Aim: SARS-CoV-2 infection is a world-wide public health problem. Several aspects of its pathogenesis and the related clinical consequences still need elucidation. In Italy, Sardinia has had very low numbers of infections. Taking advantage of the low genetic polymorphism in the Sardinian population, we analyzed clinical, genetic and immunogenetic factors, with particular attention to HLA class I and II molecules, to evaluate their influence on susceptibility to SARS-CoV-2 infection and the clinical outcome. Method and Materials: We recruited 619 healthy Sardinian controls and 182 SARS-CoV-2 patients. Thirty-nine patients required hospital care and 143 were without symptoms, pauci-symptomatic or with mild disease. For all participants, we collected demographic and clinical data and analyzed the HLA allele and haplotype frequencies. Results: Male sex and older age were more frequent in hospitalized patients, none of whom had been vaccinated during the previous seasonal flu vaccination campaignes. Compared to the group of asymptomatic or pauci-symptomatic patients, hospitalized patients also had a higher frequency of autoimmune diseases and glucose-6-phosphate-dehydrogenase (G6PDH) deficiency. None of these patients carried the beta-thalassemia trait, a relatively common finding in the Sardinian population. The extended haplotype HLA-A*02:05, B*58:01, C*07:01, DRB1*03:01 [OR 0.1 (95% CI 0–0.6), Pc = 0.015] was absent in all 182 patients, while the HLA-C*04:01 allele and the three-loci haplotype HLA-A*30:02, B*14:02, C*08:02 [OR 3.8 (95% CI 1.8–8.1), Pc = 0.025] were more frequently represented in patients than controls. In a comparison between in-patients and home care patients, the HLA-DRB1*08:01 allele was exclusively present in the hospitalized patients [OR > 2.5 (95% CI 2.7–220.6), Pc = 0.024]. Conclusion: The data emerging from our study suggest that the extended haplotype HLA-A*02:05, B*58:01, C*07:01, DRB1*03:01 has a protective effect against SARS-CoV-2 infection in the Sardinian population. Genetic factors that resulted to have a negative influence on the disease course were presence of the HLA-DRB1*08:01 allele and G6PDH deficiency, but not the beta-thalassemic trait. Absence of influenza vaccination could be a predisposing factor for more severe disease

Unveiling the gravitational universe at mu-Hz frequencies

Funding Information: Open access funding provided by Università degli Studi di Milano - Bicocca within the CRUI-CARE Agreement. Publisher Copyright: © 2021, The Author(s).We propose a space-based interferometer surveying the gravitational wave (GW) sky in the milli-Hz to mu-Hz frequency range. By the 2040s, the mu-Hz frequency band, bracketed in between the Laser Interferometer Space Antenna (LISA) and pulsar timing arrays, will constitute the largest gap in the coverage of the astrophysically relevant GW spectrum. Yet many outstanding questions related to astrophysics and cosmology are best answered by GW observations in this band. We show that a mu-Hz GW detector will be a truly overarching observatory for the scientific community at large, greatly extending the potential of LISA. Conceived to detect massive black hole binaries from their early inspiral with high signal-to-noise ratio, and low-frequency stellar binaries in the Galaxy, this instrument will be a cornerstone for multimessenger astronomy from the solar neighbourhood to the high-redshift Universe.Peer reviewe

Assessment of diagnostic criteria for multifocal motor neuropathy in patients included in the Italian database

Background and purposeThis study aimed to assess the diagnostic criteria, ancillary investigations and treatment response using real-life data in multifocal motor neuropathy (MMN) patients.MethodsClinical and laboratory data were collected from 110 patients enrolled in the Italian MMN database through a structured questionnaire. Twenty-six patients were excluded due to the unavailability of nerve conduction studies or the presence of clinical signs and symptoms and electrodiagnostic abnormalities inconsistent with the MMN diagnosis. Analyses were conducted on 73 patients with a confirmed MMN diagnosis and 11 patients who did not meet the diagnostic criteria.ResultsThe European Federation of Neurological Societies/Peripheral Nerve Society (EFNS/PNS) diagnostic criteria were variably applied. AUTHOR:When applying the American Association of Electrodiagnostic Medicine criteria, an additional 17% of patients fulfilled the criteria for probable/definite diagnosis whilst a further 9.5% missed the diagnosis. In 17% of the patients only compound muscle action potential amplitude, but not area, was measured and subsequently recorded in the database by the treating physician. Additional investigations, including anti-GM1 immunoglobulin M antibodies, cerebrospinal fluid analysis, nerve ultrasound and magnetic resonance imaging, supported the diagnosis in 46%-83% of the patients. Anti-GM1 immunoglobulin M antibodies and nerve ultrasound demonstrated the highest sensitivity. Additional tests were frequently performed outside the EFNS/PNS guideline recommendations.ConclusionsThis study provides insights into the real-world diagnostic and management strategies for MMN, highlighting the challenges in applying diagnostic criteria