SSU1 Checkup, a Rapid Tool for Detecting Chromosomal Rearrangements Related to the SSU1 Promoter in Saccharomyces cerevisiae: An Ecological and Technological Study on Wine Yeast

Chromosomal rearrangements (CR) such as translocations, duplications and inversions play a decisive role in the adaptation of microorganisms to specific environments. In enological Saccharomyces cerevisiae strains, CR involving the promoter region of the gene SSU1 lead to a higher sulfite tolerance by enhancing the SO2 efflux. To date, three different SSU1 associated CR events have been described, including translocations XV-t-XVI and VIII-t-XVI and inversion inv-XVI. In the present study, we developed a multiplex PCR method (SSU1 checkup) that allows a rapid characterization of these three chromosomal configurations in a single experiment. Nearly 600 S. cerevisiae strains collected from fermented grape juice were genotyped by microsatellite markers. We demonstrated that alleles of the SSU1 promoter are differently distributed according to the wine environment (cellar versus vineyard) and the nature of the grape juice. Moreover, rearranged SSU1 promoters are significantly enriched among commercial starters. In addition, the analysis of nearly isogenic strains collected in wine related environments demonstrated that the inheritance of these CR shapes the genetic diversity of clonal populations. Finally, the link between the nature of SSU1 promoter and the tolerance to sulfite was statistically validated in natural grape juice containing various SO2 concentrations. The SSU1 checkup is therefore a convenient new tool for addressing population genetics questions and for selecting yeast strains by using molecular markers.Fil: Marullo, Philippe. Universite de Bordeaux; FranciaFil: Claisse, Olivier. Universite de Bordeaux; FranciaFil: Raymond Eder, María Laura. Universidad Católica de Córdoba. Instituto de Investigaciones en Recursos Naturales y Sustentabilidad José Sanchez Labrador S. J. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Recursos Naturales y Sustentabilidad José Sanchez Labrador S. J.; ArgentinaFil: Börlin, Marine. Universite de Bordeaux; FranciaFil: Feghali, Nadine. Lebanese University; LíbanoFil: Bernard, Margaux. Universite de Bordeaux; FranciaFil: Legras, Jean Luc. Université Montpellier II; FranciaFil: Albertin, Warren. Universite de Bordeaux; FranciaFil: Rosa, Alberto Luis. Universidad Católica de Córdoba. Instituto de Investigaciones en Recursos Naturales y Sustentabilidad José Sanchez Labrador S. J. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Recursos Naturales y Sustentabilidad José Sanchez Labrador S. J.; ArgentinaFil: Masneuf Pomarede, Isabelle. Universite de Bordeaux; Franci

SOPROLIFE System: An Accurate Diagnostic Enhancer

Objectives. The aim of this study was to evaluate a light-emitting diode fluorescence tool, the SOPROLIFE light-induced fluorescence evaluator, and compare it to the international caries detection and assessment system-II (ICDAS-II) in the detection of occlusal caries. Methods. A total of 219 permanent posterior teeth in 21 subjects, with age ranging from 15 to 65 years, were examined. An intraclass correlation coefficient (ICC) was computed to assess the reliability between the two diagnostic methods. Results. The results showed a high reliability between the two methods (ICC = 0.92; IC = 0.901–0.940; P<0.001). The SOPROLIFE blue fluorescence mode had a high sensitivity (87%) and a high specificity (99%) when compared to ICDAS-II. Conclusion. Compared to the most used visual method in the diagnosis of occlusal caries lesions, the finding from this study suggests that SOPROLIFE can be used as a reproducible and reliable assessment tool. At a cut-off point, categorizing noncarious lesions and visual change in enamel, SOPROLIFE shows a high sensitivity and specificity. We can conclude that financially ICDAS is better than SOPROLIFE. However SOPROLIFE is easier for clinicians since it is a simple evaluation of images. Finally in terms of efficiency SOPROLIFE is not superior to ICDAS but tends to be equivalent with the same advantages

Front Microbiol

Chromosomal rearrangements (CR) such as translocations, duplications and inversions play a decisive role in the adaptation of microorganisms to specific environments. In enological strains, CR involving the promoter region of the gene lead to a higher sulfite tolerance by enhancing the SO efflux. To date, three different associated CR events have been described, including translocations XV-t-XVI and VIII-t-XVI and inversion inv-XVI. In the present study, we developed a multiplex PCR method ( checkup) that allows a rapid characterization of these three chromosomal configurations in a single experiment. Nearly 600 strains collected from fermented grape juice were genotyped by microsatellite markers. We demonstrated that alleles of the promoter are differently distributed according to the wine environment (cellar versus vineyard) and the nature of the grape juice. Moreover, rearranged promoters are significantly enriched among commercial starters. In addition, the analysis of nearly isogenic strains collected in wine related environments demonstrated that the inheritance of these CR shapes the genetic diversity of clonal populations. Finally, the link between the nature of promoter and the tolerance to sulfite was statistically validated in natural grape juice containing various SO concentrations. The checkup is therefore a convenient new tool for addressing population genetics questions and for selecting yeast strains by using molecular markers

Genetic and technological characterization of indigenous Saccharomyces cerevisiae strains isolated from 'Merwah' wine (Lebanon)

In order to gain deeper knowledge of genetic diversity and population structure of Lebanese indigenous Saccharomyces cerevisiae strains, 202 isolates were collected during spontaneous alcoholic fermentation of eight must/wine samples of ‘Merwah’, native cultivar, over two consecutive years in a traditional winery in Mount Lebanon. The isolates were identified as belonging to the S. cerevisiae species based on their morphology and preliminary analysis of the ITS 1-4 region sequence. They were then characterized at strain level by analysis of the interdelta regions, and genotyped using 12 microsatellite markers. In addition, strains were subjected to test phenotypic sulfite resistance and chromosomal translocation on SSU1gene by means of PCR. To select the most potential yeast starter strains, micro-fermentations were carried out for 22 S. cerevisiae strains that were selected as representatives of the population. Based on the main technological parameters (i.e., residual sugar, fermentative vigour, lowest sulphur production and acetic acid), three yeast strains were selected as candidates for pilot-scale fermentation at the winery. These strains were subjected to chemical and sensorial analysis for selecting a S. cerevisiae strain, adequate to produce ‘Merwah’ wine. This study will devote to test the drying attitude of the selected strain; make industrial wine using the selected dry strain ad a starter; and verify the quality of the resulting wine

Selection of Saccharomyces cerevisiae Starter Strain for Merwah Wine

In order to select Saccharomyces cerevisiae starter strains for &lsquo;&lsquo;Merwah&rsquo;&rsquo; wine production, three strains (M.6.16, M.10.16, and M.4.17) previously isolated from &lsquo;&lsquo;Merwah&rsquo;&rsquo; must and characterized at the lab scale were tested in pilot-scale fermentation in a Lebanese winery during the 2019 vintage. The three inoculated musts were compared to that obtained with a spontaneous fermentation. During the fermentations, must samples were taken to evaluate the dominance of the inoculated strains, and at the end of fermentation, the obtained wines were subjected to chemical and sensorial characterization. Molecular monitoring by interdelta analysis revealed that only M.4.17 was able to complete the fermentation and dominate over the wild yeasts. Based on the analysis of principal technological parameters (i.e., residual sugar, fermentative vigor, sulfur production, and acetic acid) and sensorial analysis of the wines obtained, M.4.17 was selected as an adequate starter for the production of typical &lsquo;&lsquo;Merwah&rsquo;&rsquo; wine

Knowledge, practice and attitudes regarding antibiotics use among Lebanese dentists

Objectives: Explore antibiotic use, assess conformity with evidence-practice guidelines, and describe knowledge and attitudinal factors among Lebanese dentists. Methods: National cross-sectional telephonic survey, using a standardized questionnaire addressing demographic, educational and professional data, usual antibiotics prophylactic and curative prescription pattern and influential factors, knowledge concerning antibiotics use in selected patient-populations, and attitude regarding antimicrobial resistance. Analyses used descriptive statistics, and bivariate analysis to observe predictors of higher knowledge. Results: the overall response rate for the study was around 21%. 322 dentists participated. On average, 17.51% of consultations resulted in antibiotic use; previous antibiotic experience mostly influenced prescriptions (81.3%). Referral of pregnant and lactating women and cardiac patients, when antibiotics are needed, was high (26.9%, 28.5% and 79.4%, respectively). Macrolides were the dominant first-line antibiotics in penicillin allergy (47.4%). Penicillins were most common for pregnant and lactating women. Penicillins (95.0%), 2g (63.9%), and 1 hour pre-procedure (34%) were the main components of prophylaxis for cardiac patients. Prophylactic and curative use varied widely; few dentists exhibited guideline-conform prescriptions. Mean knowledge scores of prophylaxis for cardiac and non-cardiac patients, and antibiotics’ side effects were predominantly poor (46.75±14.82, 39.21±33.09 and 20.27±18.77, respectively over 100). Practicing outside Beirut, undergraduate qualification in Lebanon, and post-graduate qualification predicted higher knowledge. 75.9% acknowledged the contribution of dentistry-based prescribing to antibiotic resistance and 94.7% knew at least one cause of resistance. Conclusions: Dentists show positive attitude towards antimicrobial resistance. Yet, they lack uniformity in antibiotic stewardship. Poor knowledge and guideline-incongruent prophylactic and therapeutic prescribing are observed. Development of targeted interventions is needed to promote judicious antibiotic use within Lebanese dentistry

Snail Slime Extracted by a Cruelty Free Method Preserves Viability and Controls Inflammation Occurrence: A Focus on Fibroblasts

Snail slime (SS) is a viscous secretion obtained from different snail species. SS composition is variable according to factors such as the extraction method. Even if several papers have been published regarding this topic, the molecular mechanisms at the base of SS biological effects remain unexplored. Thus, the aim of this study is to evaluate the capability of SS, extracted with the cruelty-free Muller method, to promote viability and angiogenesis processes and, in parallel, to counteract inflammation occurrence on skin cell populations. SS was administered to keratinocytes, macrophages and fibroblasts, then cell viability, through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test, cytotoxicity by lactate dehydrogenase (LDH) assay, morphology by haematoxylin-eosin staining, gene and protein expression through real-time polymerase chain reaction (PCR) and Western blot, cell cycle phases by flow cytometry, and collagen secretion using an enzyme-linked immunosorbent assay (ELISA) test, were measured. Our results evidence SS capability to promote fibroblast viability and to trigger recovery mechanisms by activating the Erk protein. Moreover, an appreciable anti-inflammatory effect due to the significant reduction in cyclooxygenase-2 expression, and a positive modulation of new blood vessel formation demonstrated by increased Angiopoietin 1 gene expression and a higher matrix deposition (evidenced by the augmented amount of released collagen I) can be identified. This evidence led us to assume that the Muller method extracted-SS represents a valuable and promising natural product suitable for cosmetic and skin care formulations

Snail Slime Extracted by a Cruelty Free Method Preserves Viability and Controls Inflammation Occurrence: A Focus on Fibroblasts

Snail slime (SS) is a viscous secretion obtained from different snail species. SS composition is variable according to factors such as the extraction method. Even if several papers have been published regarding this topic, the molecular mechanisms at the base of SS biological effects remain unexplored. Thus, the aim of this study is to evaluate the capability of SS, extracted with the cruelty-free Muller method, to promote viability and angiogenesis processes and, in parallel, to counteract inflammation occurrence on skin cell populations. SS was administered to keratinocytes, macrophages and fibroblasts, then cell viability, through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test, cytotoxicity by lactate dehydrogenase (LDH) assay, morphology by haematoxylin-eosin staining, gene and protein expression through real-time polymerase chain reaction (PCR) and Western blot, cell cycle phases by flow cytometry, and collagen secretion using an enzyme-linked immunosorbent assay (ELISA) test, were measured. Our results evidence SS capability to promote fibroblast viability and to trigger recovery mechanisms by activating the Erk protein. Moreover, an appreciable anti-inflammatory effect due to the significant reduction in cyclooxygenase-2 expression, and a positive modulation of new blood vessel formation demonstrated by increased Angiopoietin 1 gene expression and a higher matrix deposition (evidenced by the augmented amount of released collagen I) can be identified. This evidence led us to assume that the Muller method extracted-SS represents a valuable and promising natural product suitable for cosmetic and skin care formulations

Genetic and phenotypic characterisation of a saccharomyces cerevisiae population of "Merwah" white wine

The study of yeast biodiversity represents an important step in the preservation of the local heritage, and this work in particular has an innovative character since no further studies have investigated 'Merwah', one of the main grape varieties used in winemaking in Lebanon. To gain deeper knowledge of the genetic diversity and population structure of native Saccharomyces cerevisiae wine strains, 202 isolates were collected during spontaneous alcoholic fermentation of eight must/wine samples of cultivar 'Merwah', over two consecutive years (2016, 2017) in a traditional winery in Mount Lebanon (1400 m a.s.l.). The isolates were identified as S. cerevisiae on the basis of their morphology and preliminary sequence analysis of their internal transcribed spacer (ITS) PCR. They were then characterised at the strain level by interdelta PCR and genotyped using multiplex PCR reactions of 12 microsatellite markers. High genetic diversity was observed for the studied population. To select potential yeast starter strains from this population, micro-fermentations were carried out for 22 S. cerevisiae strains that were selected as representative of the 'Merwah' wine yeast population in order to determine their technological and oenological properties. Three indigenous yeast strains might represent candidates for pilot-scale fermentation in the winery, based on relevant features such as high fermentation vigour, low production of volatile acidity and H2S and low residual sugar content at the end of alcoholic fermentation