32 research outputs found

    Oral colonization by Candida species in orthodontic patients before, during and after treatment with fixed appliances : a prospective controlled trial

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    Orthodontic treatment with fixed appliances is associated with changes in oral microbiota, including increased Candida colonization. The Candida fungus can cause oral lesions and infections such as candidiasis and angular cheilitis, and is harmful to both the patient and the orthodontist. Poor hygiene facilitates the colonization of these microorganisms. The key aim was to quantify the colonization of C. albicans in patients prior to beginning orthodontic treatment, and during the treatment process. A total of 124 patients (43 males and 80 females) with a mean age of 19.5 years, who required treatment with metal or aesthetic (ceramic) braces, were studied. Microbiological samples were taken from the oral cavity using the swab technique throughout the treatment and cultured on a Sabouraud Dextrose Agar plate and, if positive, cultured on a CHROMagar® Candida plate. In contrast to other published studies, no statistically significant increase in C. albicans colonization was observed during the orthodontic treatment. The fixed appliances had no influence on the presence, absence or level of colonization by C. albicans and there were no significant differences between the different appliances studied. Our study showed that frequency of oral hygiene measures by study participants did not affect the rate of oral carriage of Candida in a statistically significant manner. This observation contrasted with published literature, which suggests that thorough brushing is important to prevent the build-up of Candida species

    Reporters for the analysis of N-glycosylation in \u3ci\u3eCandida albicans\u3c/i\u3e

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    A large proportion of Candida albicans cell surface proteins are decorated post-translationally by glycosylation. Indeed N-glycosylation is critical for cell wall biogenesis in this major fungal pathogen and for its interactions with host cells. A detailed understanding of N-glycosylation will yield deeper insights into host-pathogen interactions. However, the analysis of N-glycosylation is extremely challenging because of the complexity and heterogeneity of these structures. Therefore, in an attempt to reduce this complexity and facilitate the analysis of N-glycosylation, we have developed new synthetic C. albicans reporters that carry a single N-linked glycosylation site derived from Saccharomyces cerevisiae Suc2. These glycosylation reporters, which carry C. albicans Hex1 or Sap2 signal sequences plus carboxy-terminal FLAG3 and His6 tags, were expressed in C. albicans from the ACT1 promoter. The reporter proteins were successfully secreted and hyperglycosylated by C. albicans cells, and their outer chain glycosylation was dependent on Och1 and Pmr1, which are required for N-mannan synthesis, but not on Mnt1 and Mnt2 which are only required for O-mannosylation. These reporters are useful tools for the experimental dissection of N-glycosylation and other related processes in C. albicans, such as secretion

    Integration of antimicrobial pectin-based edible coating and active modified atmosphere packaging to preserve the quality and microbial safety of fresh-cut persimmon (Diospyros kaki Thunb. cv. Rojo Brillante)

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    BACKGROUND: The greatest hurdle to the commercial marketing of fresh-cut fruits is related to their higher susceptibility to enzymatic browning, tissue softening, and microbial growth. The aim of this study was to test the efficacy of a pectin-based edible coating and low oxygen modified atmosphere packaging (MAP) to control enzymatic browning and reduce microbial growth of fresh-cut Rojo Brillante' persimmon. The survival of Escherichia coli, Salmonella enteritidis and Listeria monocytogenes artificially inoculated on fresh-cut fruit was also assessed. The pectin coating was amended with 500 IU mL(-1) nisin (NI) as antimicrobial agent and 10gkg(-1) citric acid and 10gkg(-1) calcium chloride as anti-browning and firming agents, respectively. Persimmon slices were dipped in the coating or in water (control) and packed under 5 kPa O-2 (MAP) or in ambient atmosphere for up to 9 days at 5 degrees C. Microbial growth, package gas composition, colour, firmness, polyphenol oxidase activity, visual quality and overall sensory flavour of persimmon slices were measured during storage. RESULTS: Coating application combined with active MAP significantly reduced the CO2 emission and O-2 consumption in the package. The coating was effective in reducing browning and also inhibited the growth of mesophilic aerobic bacteria. Coating also reduced the populations of E. coli, S. enteritidis and L. monocytogenes. CONCLUSION: The combination of the pectin-based edible coating and active MAP proved to be the most effective treatment to maintain the sensory and microbiological quality of persimmon slices for more than 9 days of storage. (c) 2016 Society of Chemical Industr

    Browning inhibition and microbial control in fresh-cut persimmon (Diospyros kaki Thunb. cv. Rojo Brillante) by apple pectin-based edible coatings

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    The aim of this study was to develop new edible coatings based on apple pectin with a combination of antioxidants and antimicrobial agents to control enzymatic browning and microbial growth of fresh-cut ‘Rojo Brillante’ persimmon. The survival of important food-borne human pathogens artificially inoculated on fresh-cut fruit was also assessed. Potassium sorbate (PS) at 2 or 4 g kg−1, sodium benzoate (SB) at 4 g kg−1, or nisin (NI) at 500 IU mL−1, were added to apple pectin coatings containing 10 g kg−1 citric acid and 10 g kg−1 calcium chloride as antioxidants. Persimmon slices were dipped in the coatings, the aqueous antioxidant solution (citric acid and calcium chloride) or water (control), packed in an ambient atmosphere and stored at 5 °C for up to 9 days. Microbial growth, colour, firmness, polyphenol oxidase (PPO) activity, visual quality and overall sensory flavour were measured during storage. Coated samples and those dipped in the antioxidant aqueous solution presented lower a* values than control samples, which indicated effective browning inhibition. Persimmon slices treated with coatings containing PS and SB reached the limit of marketability after 7 days of storage. At the end of storage, the overall fruit flavour was ranked above the limit of acceptability. Antimicrobial coatings inhibited growth of mesophilic aerobic bacteria, and those containing SB and NI were the most effective. No growth of moulds, yeasts and psychrophilic aerobic bacteria was detected during storage. All the treatments effectively reduced the populations of Escherichia coli and Salmonella enteritidis, NI-coating being the most effective. For Listeria monocytogenes, only the NI-coating effectively reduced the bacterial population

    Pilot Investigation of SARS-CoV-2 Variants in the Island of Sicily Prior to and in the Second Wave of the COVID-19 Pandemic

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    8 páginas, 2 tablas, 2 figuras. The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: https://www.ncbi.nlm.nih.gov/genbank/, OM510944; https://www.ncbi.nlm.nih.gov/genbank/, OM510945; https://www.ncbi. nlm.nih.gov/genbank/, OM510946; https://www.ncbi.nlm.nih. gov/genbank/, OM510947; https://www.ncbi.nlm.nih.gov/genbank/, OM510948; https://www.ncbi.nlm.nih.gov/genbank/, OM510949; https://www.ncbi.nlm.nih.gov/genbank/, OM51 0950; https://www.ncbi.nlm.nih.gov/genbank/, OM510951; and https://www.ncbi.nlm.nih.gov/genbank/, OM510952.After 2 years of the COVID-19 pandemic, we continue to face vital challenges stemming from SARS-CoV-2 variation, causing changes in disease transmission and severity, viral adaptation to animal hosts, and antibody/vaccine evasion. Since the monitoring, characterization, and cataloging of viral variants are important and the existing information on this was scant for Sicily, this pilot study explored viral variants circulation on this island before and in the growth phase of the second wave of COVID-19 (September and October 2020), and in the downslope of that wave (early December 2020) through sequence analysis of 54 SARS-CoV-2-positive samples. The samples were nasopharyngeal swabs collected from Sicilian residents by a state-run one-health surveillance laboratory in Palermo. Variant characterization was based on RT-PCR amplification and sequencing of four regions of the viral genome. The B.1.177 variant was the most prevalent one, strongly predominating before the second wave and also as the wave downsized, although its relative prevalence decreased as other viral variants, particularly B.1.160, contributed to virus circulation. The occurrence of the B.1.160 variant may have been driven by the spread of that variant in continental Europe and by the relaxation of travel restrictions in the summer of 2020. No novel variants were identified. As sequencing of the entire viral genome in Sicily for the period covered here was restricted to seven deposited viral genome sequences, our results shed some light on SARS-CoV-2 variant circulation during that wave in this insular region of Italy which combines its partial insular isolation with being a major entry point for the African immigration.This research received external funding to CR-G and EM from “Agencia Valenciana de Innovación: COVID-19, Ayudas de Concesión Directa a Soluciones Científico-Innovadoras Directamente Relacionadas Con La Lucha Contra La COVID-19,” (Ref COVID-19-203) and from “Conselleria de Innovación Universidades, Ciencia y Sociedad Digital: Subvenciones a Grupos de Investigación Emergentes” (Ref, GV/2021/163); to VR from the Agencia Estatal de Investigación of the Spanish Government (Ref PID2020-120322RB-C21) and from the European Commission–NextGeneration EU CSIC Global Health Platform, Spanish Ministry of Science and Innovation (Ref MCIN/AEI/10.13039/501100011033); and to the Istituto Zooprofilattico Sperimentale della Sicilia “A. Mirri” by the Project “COVID-19: Traiettorie Evolutive di SARS-CoV-2 ed Indagine Sul Ruolo Degli Animali” (IZS SI 03/20 RC), funded by the Italian Ministry of Health.Peer reviewe

    A SARS-CoV-2 full genome sequence of the B.1.1 lineage sheds light on viral evolution in Sicily in late 2020

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    9 páginas, 2 figuras, 2 tablasTo investigate the influence of geographic constrains to mobility on SARS-CoV-2 circulation before the advent of vaccination, we recently characterized the occurrence in Sicily of viral lineages in the second pandemic wave (September to December 2020). Our data revealed wide prevalence of the then widespread through Europe B.1.177 variant, although some viral samples could not be classified with the limited Sanger sequencing tools used. A particularly interesting sample could not be fitted to a major variant then circulating in Europe and has been subjected here to full genome sequencing in an attempt to clarify its origin, lineage and relations with the seven full genome sequences deposited for that period in Sicily, hoping to provide clues on viral evolution. The obtained genome is unique (not present in databases). It hosts 20 single-base substitutions relative to the original Wuhan-Hu-1 sequence, 8 of them synonymous and the other 12 encoding 11 amino acid substitutions, all of them already reported one by one. They include four highly prevalent substitutions, NSP12:P323L, S:D614G, and N:R203K/G204R; the much less prevalent S:G181V, ORF3a:G49V and N:R209I changes; and the very rare mutations NSP3:L761I, NSP6:S106F, NSP8:S41F and NSP14:Y447H. GISAID labeled this genome as B.1.1 lineage, a lineage that appeared early on in the pandemic. Phylogenetic analysis also confirmed this lineage diagnosis. Comparison with the seven genome sequences deposited in late 2020 from Sicily revealed branching leading to B.1.177 in one branch and to Alpha in the other branch, and suggested a local origin for the S:G118V mutation.This research received external funding to CR-G from Conselleria de Innovación Universidades, Ciencia y Sociedad Digital: Subvenciones a Grupos de Investigación Emergentes (Ref, GV/2021/163); to VR from the Agencia Estatal de Investigación of the Spanish Government (Ref PID2020-120322RB-C21) and the European Commission-NextGeneration EU CSIC Global Health Platform, Spanish Ministry of Science and Innovation (Ref MCIN/AEI/10.13039/501100011033); and to the Istituto Zooprofilattico Sperimentale della Sicilia A. Mirri by the Project COVID-19: Traiettorie Evolutive di SARS-CoV-2 ed Indagine Sul Ruolo Degli Animali (IZS SI 03/20 RC), funded by the Italian Ministry of HealthPeer reviewe

    Glycosylation status of the C. albicans cell wall affects the efficiency of neutrophil phagocytosis and killing but not cytokine signaling

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    The cell wall of the opportunistic human fungal pathogen, Candida albicans is a complex, layered network of rigid structural polysaccharides composed of β-glucans and chitin that is covered with a fibrillar matrix of highly glycosylated mannoproteins. Poly-morphonuclear cells (PMNs, neutrophils) are the most prevalent circulating phagocytic leukocyte in peripheral blood and they are pivotal in the clearance of invading fungal cells from tissues. The importance of cell-wall mannans for the recognition and uptake of C. albicans by human PMNs was therefore investigated. N- and O-glycosylation-deficient mutants were attenuated in binding and phagocytosis by PMNs and this was associated with reduced killing of C. albicans yeast cells. No differences were found in the production of the respiratory burst enzyme myeloperoxidase (MPO) and the neutrophil chemokine IL-8 in PMNs exposed to control and glycosylation-deficient C. albicans strains. Thus, the significant decrease in killing of glycan-deficient C. albicans strains by PMNs is a consequence of a marked reduction in phagocytosis rather than changes in the release of inflammatory mediators by PMNs

    Molecular characterisation of CO2-mediated polymorphism in Candida albicans

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