siRNA that participates in Drosophila dosage compensation is produced by many 1.688X and 359†bp repeats.

Organisms with differentiated sex chromosomes must accommodate unequal gene dosage in males and females. Male fruit flies increase X-linked gene expression to compensate for hemizygosity of their single X chromosome. Full compensation requires localization of the Male-Specific Lethal (MSL) complex to active genes on the male X, where it modulates chromatin to elevate expression. The mechanisms that identify X chromatin are poorly understood. The euchromatic X is enriched for AT-rich, ∼359 bp satellites termed the 1.688X repeats. Autosomal insertions of 1.688X DNA enable MSL recruitment to nearby genes. Ectopic expression of dsRNA from one of these repeats produces siRNA and partially restores X-localization of MSLs in males with defective X recognition. Surprisingly, expression of double-stranded RNA from three other 1.688X repeats failed to rescue males. We reconstructed dsRNA-expressing transgenes with sequence from two of these repeats and identified phasing of repeat DNA, rather than sequence or orientation, as the factor that determines rescue of males with defective X recognition. Small RNA sequencing revealed that siRNA was produced in flies with a transgene that rescues, but not in those carrying a transgene with the same repeat but different phasing. We demonstrate that pericentromeric X heterochromatin promotes X recognition through a maternal effect, potentially mediated by small RNA from closely related heterochromatic repeats. This suggests that the sources of siRNAs promoting X recognition are highly redundant. We propose that enrichment of satellite repeats on Drosophilid X chromosomes facilitates the rapid evolution of differentiated sex chromosomes by marking the X for compensation.

Virulence factors in multidrug (MDR) and Pan-drug resistant (XDR) Pseudomonas aeruginosa: a cross-sectional study of isolates recovered from ocular infections in a high-incidence setting in southern India.

BACKGROUND: Global concerns have been raised due to upward trend of Multi-drug Resistant (MDR) Pseudomonas aeruginosa reports in ocular infections. Our aim was to characterize the virulence determinants of MDR P. aeruginosa causing ocular infections. METHODS: P. aeruginosa strains were isolated from 46 patients with conjunctivitis (2), endophthalmitis (11) and active keratitis (25) seen at our Institute, between 2016 and 2020. The isolates were identified by Vitek-2 and characterized based on growth kinetics, biofilm formation, motility, pyoverdine and pyocyanin production, phospholipase and catalase activity, urease production along with expression of exotoxins (exo-A, exo-U and exo-S) and correlated to its antibiotic profiles. RESULTS: Of the 46 P. aeruginosa isolates, 23 were MDR and were significantly (p = 0.03) associated with older (> 65) patients, along with higher production of pyoverdine (58.3%), pyocyanin (30.4%), phospholipase (91.6%) and protease (62.5%) activity, formed strong biofilms and exo-A (30.4%). No significant relation between motility, urease and catalase production with antibiotic susceptibility was observed. Heatmap and PCoA analysis confirmed this unique virulence profile associated with MDR-PA strains. CONCLUSION: Phenotypic characteristics of P.aeruginosa might be responsible for increased colonization and antibiotic resistance observed in vivo and understanding these differences may lead to development of clinical guidelines for the management of MDR infections.