Role of KLHL3 and dietary K⁺ in regulating KS-WNK1 expression

This is the author accepted manuscript. The final version is available from the American Physiological Society via the DOI in this recordThe physiological role of the shorter isoform of WNK1 that is exclusively expressed in the kidney (KS-WNK1), with particular abundance in the distal convoluted tubule, remains elusive. KS-WNK1 despite lacking the kinase domain, is nevertheless capable of stimulating the NaCl cotransporter (NCC), apparently through activation of WNK4. It has recently been shown that a less severe form of the Familial Hyperkalemic Hypertension featuring only hyperkalemia is caused by missense mutations in the WNK1 acidic domain that preferentially affect CUL3-KLHL3 E3-induced degradation of KS-WNK1, rather than that of the full-length WNK1 (L-WNK1). Here we show that L-WNK1 is indeed less impacted by the CUL3-KLHL3 E3 ligase complex compared to KS-WNK1. We demonstrate that the unique 30 amino acid amino N-terminal fragment of KS-WNK1 is essential for its activating effect on NCC and recognition by KLHL3. We identify specific amino acid residues in this region critical for the functional effect of KS-WNK1 and KLHL3 sensitivity. To further explore this, we generated KLHL3-R528H knock-in mice that mimic human mutations causing Familial Hyperkalemic Hypertension. These mice revealed that the KLHL3 mutation specifically increased expression of KS-WNK1 in the kidney. We also observed that in wild type mice, expression of KS-WNK1 is only detectable after exposure to low potassium diet. These findings provide new insights into the regulation and function of KS-WNK1 by the CUL3-KLHL3 complex in DCT and indicate that this pathway is regulated by dietary K+ levels.National Institutes of Health (NIH)Conacyt MexicoPAPIIT UNAML'OréalMedical Research Council (MRC

Systems biology analysis of temporal in vivo Brucella melitensis and bovine transcriptomes predicts host:pathogen protein-protein interactions

To date, fewer than 200 gene-products have been identified as Brucella virulence factors, and most were characterized individually without considering how they are temporally and coordinately expressed or secreted during the infection process. Here, we describe and analyze the in vivo temporal transcriptional profile of Brucella melitensis during the initial 4 h interaction with cattle. Pathway analysis revealed an activation of the “Two component system” providing evidence that the in vivo Brucella sense and actively regulate their metabolism through the transition to an intracellular lifestyle. Contrarily, other Brucella pathways involved in virulence such as “ABC transporters” and “T4SS system” were repressed suggesting a silencing strategy to avoid stimulation of the host innate immune response very early in the infection process. Also, three flagellum-encoded loci (BMEII0150-0168, BMEII1080-1089, and BMEII1105-1114), the “flagellar assembly” pathway and the cell components “bacterial-type flagellum hook” and “bacterial-type flagellum” were repressed in the tissue-associated B. melitensis, while RopE1 sigma factor, a flagellar repressor, was activated throughout the experiment. These results support the idea that Brucella employ a stealthy strategy at the onset of the infection of susceptible hosts. Further, through systems-level in silico host:pathogen protein–protein interactions simulation and correlation of pathogen gene expression with the host gene perturbations, we identified unanticipated interactions such as VirB11::MAPK8IP1; BtaE::NFKBIA, and 22 kDa OMP precursor::BAD and MAP2K3. These findings are suggestive of new virulence factors and mechanisms responsible for Brucella evasion of the host's protective immune response and the capability to maintain a dormant state. The predicted protein–protein interactions and the points of disruption provide novel insights that will stimulate advanced hypothesis-driven approaches toward revealing a clearer understanding of new virulence factors and mechanisms influencing the pathogenesis of brucellosis.To date, fewer than 200 gene-products have been identified as Brucella virulence factors, and most were characterized individually without considering how they are temporally and coordinately expressed or secreted during the infection process. Here, we describe and analyze the in vivo temporal transcriptional profile of Brucella melitensis during the initial 4 h interaction with cattle. Pathway analysis revealed an activation of the "Two component system" providing evidence that the in vivo Brucella sense and actively regulate their metabolism through the transition to an intracellular lifestyle. Contrarily, other Brucella pathways involved in virulence such as "ABC transporters" and "T4SS system" were repressed suggesting a silencing strategy to avoid stimulation of the host innate immune response very early in the infection process. Also, three flagellum-encoded loci (BMEII0150-0168, BMEII1080-1089, and BMEII1105-1114), the "flagellar assembly" pathway and the cell components "bacterial-type flagellum hook" and "bacterial-type flagellum" were repressed in the tissue-associated B. melitensis, while RopE1 sigma factor, a flagellar repressor, was activated throughout the experiment. These results support the idea that Brucella employ a stealthy strategy at the onset of the infection of susceptible hosts. Further, through systems-level in silico host:pathogen protein-protein interactions simulation and correlation of pathogen gene expression with the host gene perturbations, we identified unanticipated interactions such as VirB11Peer reviewedVeterinary Pathobiolog

Supraglottic airway devices for blind endotracheal intubation: A systematic review.

IntroductionThe effectiveness of supraglottic airway devices (SGDs) as a strategy for blind endotracheal intubation (ETI) was compared in this study.MethodsA systematic review of clinical trials (CTs) involving SGDs for blind ETI in patients under general anesthesia or simulation manikins, was conducted. CTs that used SGDs for fiberoptic-guided ETI and those conducted in children were excluded. Searches were performed in Embase, MEDLINE (PubMed), Scopus, and LILACS. The primary outcomes examined were the success rate of blind ETI and intubation time. Secondary outcomes were first-attempt intubation success rate and perceived ease of use.ResultsA total of 567 records were identified from databases, and 16 were identified through citation searches. Ultimately, 27 CTs met the inclusion criteria. The Fastrach Intubating Laryngeal Mask Airway (LMA Fastrach), i-gel, Air-Q Intubating Laryngeal Airway, and Supraglottic Airway Laryngopharyngeal Tube (S.A.L.T.) were the most used SGDs for blind ETI. LMA Fastrach was the most frequently compared device in these CTs. Among the studies in patients, LMA Fastrach and i-gel were the devices that showed the shortest intubation time, although it may be influenced by the way intubation time is assessed. The SGDs with the highest overall success rate were i-gel, S.A.L.T., LMA Fastrach, and single-use LMA Fastrach, followed by Air-Q, and the Intubating Laryngeal Tube Suction-Disposable (iLTS-D2), all achieving success rates greater than 90%. AuraGain had the lowest first-attempt and overall success rates for blind ETI with SGDs.ConclusionNew SGDs have not surpassed the LMA Fastrach effectiveness for blind ETI. The single-use LMA Fastrach combines the efficacy of the reusable LMA Fastrach with the features of other SGDs and may be a suitable replacement for them. The I-gel is also a viable alternative for blind ETI, while the AuraGain may not be recommended for this purpose