The million mutation project: a new approach to genetics in Caenorhabditis elegans.

We have created a library of 2007 mutagenized Caenorhabditis elegans strains, each sequenced to a target depth of 15-fold coverage, to provide the research community with mutant alleles for each of the worm's more than 20,000 genes. The library contains over 800,000 unique single nucleotide variants (SNVs) with an average of eight nonsynonymous changes per gene and more than 16,000 insertion/deletion (indel) and copy number changes, providing an unprecedented genetic resource for this multicellular organism. To supplement this collection, we also sequenced 40 wild isolates, identifying more than 630,000 unique SNVs and 220,000 indels. Comparison of the two sets demonstrates that the mutant collection has a much richer array of both nonsense and missense mutations than the wild isolate set. We also find a wide range of rDNA and telomere repeat copy number in both sets. Scanning the mutant collection for molecular phenotypes reveals a nonsense suppressor as well as strains with higher levels of indels that harbor mutations in DNA repair genes and strains with abundant males associated with him mutations. All the strains are available through the Caenorhabditis Genetics Center and all the sequence changes have been deposited in WormBase and are available through an interactive website.

Safe and effective protocol for discharge 3 days after cardiac surgery.

The Enhanced Recovery After Surgery (ERAS) protocol affected traditional cardiac surgery processes and COVID-19 is expected to accelerate its scalability. The aim of this study was to assess the impact of an ERAS-based protocol on the length of hospital stay after cardiac surgery. From January 2019 to June 2020, 664 patients underwent consecutive cardiac surgery at a Latin American center. Here, 46 patients were prepared for a rapid recovery through a multidisciplinary institutional protocol based on the ERAS concept, the "TotalCor protocol". After the propensity score matching, 46 patients from the entire population were adjusted for 12 variables. Patients operated on the TotalCor protocol had reduced intensive care unit time (P < 0.025), postoperative stay (P ≤ 0.001) and length of hospital stay (P ≤ 0.001). In addition, there were no significant differences in the occurrence of complications and death between the two groups. Of the 10-central metrics of TotalCor protocol, 6 had > 70% adherences. In conclusion, the TotalCor protocol was safe and effective for a 3-day discharge after cardiac surgery. Postoperative atrial fibrillation and renal failure were predictors of postoperative stay > 5 days.

Discrepancy Between Internal and External Intracranial Pressure Transducers: Quantification of an Old Source of Error in EVDs?

BACKGROUND: Intracranial pressure monitoring remains the foundation for prevention of secondary injury after traumatic brain injury and is most commonly performed using an external ventricular drain or intraparenchymal pressure monitor. The Integra Flex ventricular catheter combines an external ventricular catheter with a pressure transducer embedded in the tip of the catheter to allow continuous pressure readings while simultaneously draining cerebrospinal fluid. Discrepancies between measurements from the continuously reported internal pressure transducer and intermittently assessed and externally transduced ventricular drain prompted an analysis and characterization of pressures transduced from the same ventricular source. METHODS: More than 500 hours of high-resolution (125 Hz) continuous recordings were manually reviewed to identify 73 hours of simultaneous measurements (clamped external ventricular drain) from internal and external transducers in patients with traumatic brain injury. RESULTS: A significant positive bias was found in pressure readings obtained from external relative to internal measurements. The 2 methods of measurement generally correlated poorly with each other and variably. Although proportional bias was found with Bland-Altman analysis, coherence revealed rare shifts in the external transducer as a major source of discrepancy. Infrequent changes in the 0-level of the external transducer were found to be the primary source of discrepancy. Relative to the observed differences, no significant trend was observed over time between the 2 modalities. CONCLUSIONS: This study suggests that the internal pressure transducer may be a more reliable estimate of intracranial pressure relative to bedside external transducers due to the inherent behavioral requirement of leveling.

A novel anti-mycobacterial function of mitogen-activated protein kinase phosphatase-1.

BACKGROUND: Mycobacterium tuberculosis (MTB) is a major cause of morbidity and mortality in the world. To combat against this pathogen, immune cells release cytokines including tumor necrosis factor-alpha (TNF-alpha), which is pivotal in the development of protective granulomas. Our previous results showed that Bacillus Calmette Guerin (BCG), a mycobacterium used as a model to investigate the immune response against MTB, stimulates the induction of TNF-alpha via mitogen-activated protein kinase (MAPK) in human blood monocytes. Since MAPK phosphatase-1 (MKP-1) is known to regulate MAPK activities, we examined whether MKP-1 plays a role in BCG-induced MAPK activation and cytokine expression. RESULTS: Primary human blood monocytes were treated with BCG and assayed for MKP-1 expression. Our results demonstrated that following exposure to BCG, there was an increase in the expression of MKP-1. Additionally, the induction of MKP-1 was regulated by p38 MAPK and extracellular signal-regulated kinase 1 and 2 (ERK1/2). Surprisingly, when MKP-1 expression was blocked by its specific siRNA, there was a significant decrease in the levels of phospho-MAPK (p38 MAPK and ERK1/2) and TNF-alpha inducible by BCG. CONCLUSIONS: Since TNF-alpha is pivotal in granuloma formation, the results indicated an unexpected positive function of MKP-1 against mycobacterial infection as opposed to its usual phosphatase activity.