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OD600nm readings of BCG0642c significantly differed in days 0, 2, 4, and from day 10 to day 12 (stationary phase) (Fig. 3b); however, doubling time differed at day 1 (BCG::pMV361, 20.72 ± 1.33 h vs. BCG::BCG0642c, 29.05 ± 2 h; p = 0.0181) and day 3 only (BCG::pMV361, 21 ± 0.76 h vs. BCG::BCG0642c, 14.3 ± 1.97 h; p = 0.0067). hsp60-driven expression of BCG3766c resulted in significant differences in OD600nm readings at day 2, and from day 6 to day 8 (mid-log phase), with doubling time being significantly different at days 2 and 3 of culture (day 2, BCG::pMV361, 20.72 ± 1.33 h vs. BCG::BCG3766c, 26.08 ± 1.29 h; p = 0.0065. Day 3, BCG::pMV361, 21 ± 0.76 h vs. BCG::BCG3766c, 18.19 ± 0.75 h; p = 0.0111). Pedroza-Roldan, C. et al. BCG constitutively expressing the adenylyl cyclase encoded by Rv2212 increases its immunogenicity and reduces replication of M. tuberculosis in lungs of BALB/c mice. Tuberculosis (Edinb) 113, 19–25. https://doi.org/10.1016/j.tube.2018.08.012 (2018). Pacl, H. T., Reddy, V. P., Saini, V., Chinta, K. C. & Steyn, A. J. C. Host-pathogen redox dynamics modulate Mycobacterium tuberculosis pathogenesis. Pathog. Dis. https://doi.org/10.1093/femspd/fty036 (2018). To complete our gene expression analyses, we used RT-qPCR to monitor the expression of BCG0114, dosR, BCG0642c, ethR, and BCG3766c in planktonic cultures of BCG at early-log and stationary phase (Fig. 1b). Using early-log phase cultures as a reference, we observed that expression of all genes in stationary cultures followed the same pattern as they did during intercellular aggregation ( BCG0114 and dosR upregulated; BCG0642c, ethR, and BCG3766c downregulated). Hence, RT-qPCR validated differential expression for 4 out of 5 genes selected from RNA-Seq assays. Moreover, it showed that expression of the 5 selected targets in stationary phase planktonic cultures resembled the pattern found during intercellular aggregation. Phenotypic changes in multicellular BCG aggregates derived from increased expression of dosR, BCG0114, BCG0642c, ethR, and BCG3766c Sono la dottoressa Monica Greco di Rivalta di Torino, medico chirurgo specialista in Fisiatria e Geriatria ed esperta in alimentazione grazie a plurimi Master conseguiti nel corso degli anni di studi (Dieta GIFT, RIZA, Medicina Tradizionale Cinese ed auricoloterapia, STARGUO).

Lefebvre, C. et al. Discovery of a novel dehydratase of the fatty acid synthase type II critical for ketomycolic acid biosynthesis and virulence of Mycobacterium tuberculosis. Sci. Rep. 10, 2112. https://doi.org/10.1038/s41598-020-58967-8 (2020).Sono stati 28 giorni bellissimi , pieni di emozioni perché vivevamo la trasformazione dentro di noi e in più avevamo la voglia di esplorare ogni giorno una ricetta nuova. But what does it mean to become a World Surfing Reserve? Climate change and overdevelopment threatens to disrupt the carefully balanced geophysical ecosystem at these surf spots. If the cliffs, dunes and sea bed shift significantly, the swell cannot shape into the perfect crescent waves needed to surf. By achieving this protected status, local surfers are given a more prominent voice when it comes to challenging decision makers on environmental matters affecting the coastline. This ranges from fighting sewage dumping in the ocean, safeguarding the shoreline from encroaching development, and conserving wild spaces for wildlife such as bottlenose dolphins, grey seals and basking sharks. In north Devon this will give surf spots comparable levels of protection to nearby Dartmoor and Exmoor national parks. The area’s surfing history and culture was also a key feature in the award of WSR status. Prabhakara, R., Harro, J. M., Leid, J. G., Harris, M. & Shirtliff, M. E. Murine immune response to a chronic Staphylococcus aureus biofilm infection. Infect. Immun. 79, 1789–1796. https://doi.org/10.1128/IAI.01386-10 (2011). Anderson, G. G. & O’Toole, G. A. Innate and induced resistance mechanisms of bacterial biofilms. Curr. Top. Microbiol. Immunol. 322, 85–105. https://doi.org/10.1007/978-3-540-75418-3_5 (2008).

Mycobacterium tuberculosis and M. smegmatis form drug-tolerant biofilms through dedicated genetic programs 6, 7. In support of a stepwise process regulating biofilm production in mycobacteria, it was recently shown that in M. smegmatis, lsr2 participates in intercellular aggregation, while groEL1 was required for biofilm maturation 1. Additionally, it was found that multiple genes that are necessary for fitness of M. tuberculosis cells within biofilms, were also implicated in tolerance to a diverse set of stressors and antibiotics 8, something not observed for planktonic cells, further supporting a role for M. tuberculosis biofilms in drug tolerance 7. Twelve SNPs in genes (seven non-synonymous and five synonymous) and two SNPs in the intergenic region were identified as two consecutive nucleotide substitutions. Three SNPs were shared among all samples. CSBF and CMBF had the lowest number of SNPs and CSBL had the highest. A non-synonymous SNP in RS01830 encoding a glycerophosphoryl diester phosphodiesterase enzyme was only detected in CSPL and CSBL and may be related to the number of passages. Six SNPs in RS10645 were detected in CSPM, CSPL, CSBL and CMBL. This gene is involved in the polyketide and possibly phytoserol synthesis, and also shows variants in BCG strains compared to the M. tuberculosis H37Rv.Per molti anni in famiglia siamo stati tutti vegetariani, sentivo però che mancava qualcosa , che non tutti i tasselli erano messi al posto giusto finché, navigando sul web, ho conosciuto Francesca. The last gene we evaluated was BCG3766c, which encodes for a conserved hypothetical proline rich protein (Supplementary Table 1). This gene was significantly downregulated during surface attachment and was also downregulated during the intercellular aggregation step as well (FC 0.76, p = 0.022). This may explain why its expression from hsp60 tended to reduce biofilm production (Fig. 2). Hengge, R. Linking bacterial growth, survival, and multicellularity: small signaling molecules as triggers and drivers. Curr. Opin. Microbiol. 55, 57–66. https://doi.org/10.1016/j.mib.2020.02.007 (2020). We also observed changes in mature surface pellicle appearance formed in tissue culture flasks, as both BCG:: BCG0114 and BCG:: dosR produced smooth pellicles as opposed to the rugose ones produced by BCG::pMV361, BCG:: ethR, BCG:: BCG0642c, and BCG:: BCG3766c, with slight variations among strains in apparent rugosity (Fig. 2b). Beware of offshore winds if you are using inflatables, paddle boards or kayaks. These winds can blow you