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Increased plasma mitochondrial DNA concentrations are associated with poor outcomes in multiple critical illnesses, including COVID-19. However, current methods of cell-free mitochondrial DNA quantification in plasma are time-consuming and lack reproducibility. Here, we used next-generation sequencing to characterize the size and genome location of circulating mitochondrial DNA in critically ill subjects with COVID-19 to develop a facile and optimal method of quantification by droplet digital PCR. Sequencing revealed a large percentage of small mitochondrial DNA fragments in plasma with wide variability in coverage by genome location. We identified probes for the mitochondrial DNA genes, cytochrome B and NADH dehydrogenase 1, in regions of relatively high coverage that target small sequences potentially missed by other methods. Serial assessments of absolute mitochondrial DNA concentrations were then determined in plasma from 20 critically ill subjects with COVID-19 without a DNA isolation step. Mitochondrial DNA concentrations on the day of enrollment were increased significantly in patients with moderate or severe acute respiratory distress syndrome (ARDS) compared with those with no or mild ARDS. Comparisons of mitochondrial DNA concentrations over time between patients with no/mild ARDS who survived, patients with moderate/severe ARDS who survived, and nonsurvivors showed the highest concentrations in patients with more severe disease. Absolute mitochondrial DNA quantification by droplet digital PCR is time-efficient and reproducible; thus, we provide a valuable tool and rationale for future studies evaluating mitochondrial DNA as a real-time biomarker to guide clinical decision-making in critically ill subjects with COVID-19. [ABSTRACT FROM AUTHOR]

Keywords: giant prostatic calculi; moses technology; moses lithotripsy; dystrophic calcification; giant prostatic calculus; radiotherapy of prostate; #EndoUrology; #UroStone; #PCSM; #ProstateCancer; #uroonc EN giant prostatic calculi moses technology moses lithotripsy dystrophic calcification giant prostatic calculus radiotherapy of prostate #EndoUrology #UroStone #PCSM #ProstateCancer #uroonc 306 308 3 03/22/22 20220301 NES 220301 Abbreviations DC dystrophic calcification RT radiotherapy TURP transurethral resection of the prostate Case Presentation A 74-year-old man with a history of prostate cancer, nephrolithiasis, recurrent urinary retention presented to clinic with 2 years of pelvic discomfort and dysuria and a 1 week history of new onset haematuria and urethralgia. Giant prostatic calculi, moses technology, moses lithotripsy, dystrophic calcification, giant prostatic calculus, radiotherapy of prostate, #EndoUrology, #PCSM, #ProstateCancer, #uroonc, #UroStone. [Extracted from the article]

Rab proteins are conserved small GTPases that coordinate intracellular trafficking essential to cellular function and homeostasis. RAB-6.2 is a highly conserved C. elegans ortholog of human RAB6 proteins. RAB-6.2 is expressed in most tissues in C. elegans and is known to function in neurons and in the intestine tomediate retrograde trafficking. Here,we showthatRAB-6.2 is necessary for cuticle integrity and impermeability in C. elegans. RAB-6.2 functions in the epidermis to instruct skin integrity. Significantly, we show that expression of a mouse RAB6A cDNA can rescue defects in C. elegans epidermis caused by lack of RAB-6.2, suggesting functional conservation across phyla. We also show that the novel function of RAB-6.2 in C. elegans cuticle development is distinct from its previously described function in neurons. Exocyst mutants partially phenocopy rab-6.2-null animals, and rab-6.2-null animals phenocopy mutants that have defective surface glycosylation. These results suggest that RAB-6.2 may mediate the trafficking of one or many secreted glycosylated cuticle proteins directly, or might act indirectly by trafficking glycosylation enzymes to their correct intracellular localization. [ABSTRACT FROM AUTHOR]

Rab proteins are conserved small GTPases that coordinate intracellular trafficking essential to cellular function and homeostasis. RAB-6.2 is a highly conserved C. elegans ortholog of human RAB6. RAB-6.2 is expressed in most tissues in C. elegans and is known to function in neurons and in the intestine to mediate retrograde trafficking. Here we show that RAB-6.2 is necessary for cuticle integrity and impermeability in C. elegans. RAB-6.2 functions in the epidermis to instruct skin integrity. Significantly, we show that a mouse RAB6A cDNA can rescue defects in C. elegans epidermis suggesting functional conservation across phyla. We also show that the novel function of RAB-6.2 in C. elegans cuticle development is distinct from its previously described function in neurons. Exocyst mutants partially phenocopy rab-6.2 null animals and rab-6.2 null animals phenocopy mutants that have defective surface glycosylation. These results suggest that RAB-6.2 may be mediating the trafficking of one or many secreted glycosylated cuticle proteins directly, or indirectly by trafficking glycosylation enzymes to their correct intracellular localization. [ABSTRACT FROM AUTHOR]