Loss-of-function mutations in Lysyl-tRNA synthetase cause various leukoencephalopathy phenotypes

Objective: To expand the clinical spectrum of lysyl-tRNA synthetase (KARS) gene–related diseases, which so far includes Charcot-Marie-Tooth disease, congenital visual impairment and microcephaly, and nonsyndromic hearing impairment. Methods: Whole-exome sequencing was performed on index patients from 4 unrelated families with leukoencephalopathy. Candidate pathogenic variants and their cosegregation were confirmed by Sanger sequencing. Effects of mutations on KARS protein function were examined by aminoacylation assays and yeast complementation assays. Results: Common clinical features of the patients in this study included impaired cognitive ability, seizure, hypotonia, ataxia, and abnormal brain imaging, suggesting that the CNS involvement is the main clinical presentation. Six previously unreported and 1 known KARS mutations were identified and cosegregated in these families. Two patients are compound heterozygous for missense mutations, 1 patient is homozygous for a missense mutation, and 1 patient harbored an insertion mutation and a missense mutation. Functional and structural analyses revealed that these mutations impair aminoacylation activity of lysyl-tRNA synthetase, indicating that de- fective KARS function is responsible for the phenotypes in these individuals. Conclusions: Our results demonstrate that patients with loss-of-function KARS mutations can manifest CNS disorders, thus broadening the phenotypic spectrum associated with KARS-related disease

Chemical Genetic Analysis and Functional Characterization of Staphylococcal Wall Teichoic Acid 2-Epimerases Reveals Unconventional Antibiotic Drug Targets

Here we describe a chemical biology strategy performed in Staphylococcus aureus and Staphylococcus epidermidis to identify MnaA, a 2-epimerase that we demonstrate interconverts UDP-GlcNAc and UDP-ManNAc to modulate substrate levels of TarO and TarA wall teichoic acid (WTA) biosynthesis enzymes. Genetic inactivation of mnaA results in complete loss of WTA and dramatic in vitro β-lactam hypersensitivity in methicillin-resistant S. aureus (MRSA) and S. epidermidis (MRSE). Likewise, the β-lactam antibiotic imipenem exhibits restored bactericidal activity against mnaA mutants in vitro and concomitant efficacy against 2-epimerase defective strains in a mouse thigh model of MRSA and MRSE infection. Interestingly, whereas MnaA serves as the sole 2-epimerase required for WTA biosynthesis in S. epidermidis, MnaA and Cap5P provide compensatory WTA functional roles in S. aureus. We also demonstrate that MnaA and other enzymes of WTA biosynthesis are required for biofilm formation in MRSA and MRSE. We further determine the 1.9Å crystal structure of S. aureus MnaA and identify critical residues for enzymatic dimerization, stability, and substrate binding. Finally, the natural product antibiotic tunicamycin is shown to physically bind MnaA and Cap5P and inhibit 2-epimerase activity, demonstrating that it inhibits a previously unanticipated step in WTA biosynthesis. In summary, MnaA serves as a new Staphylococcal antibiotic target with cognate inhibitors predicted to possess dual therapeutic benefit: as combination agents to restore β-lactam efficacy against MRSA and MRSE and as non-bioactive prophylactic agents to prevent Staphylococcal biofilm formation.publishe

Round-robin study for ice adhesion tests

Ice adhesion tests are widely used to assess the performance of potential icephobic surfaces and coatings. A great variety of test designs have been developed and used over the past decades due to the lack of formal standards for these types of tests. In many cases, the aim of the research was not only to determine ice adhesion values, but also to understand the key surface properties correlated to low ice adhesion surfaces. Data from different measurement techniques had low correspondence between the results: Values varied by orders of magnitude and showed different relative relationships to one another. This study sought to provide a broad comparison of ice adhesion testing approaches by conducting different ice adhesion tests with identical test surfaces. A total of 15 test facilities participated in this round-robin study, and the results of 13 partners are summarized in this paper. For the test series, ice types (impact and static) as well as test parameters were harmonized to minimize the deviations between the test setups. Our findings are presented in this paper, and the ice- and test-specific results are discussed. This study can improve our understanding of test results and support the standardization process for ice adhesion strength measurements

Effect of Actual Gas Turbine Operating Conditions on Mist/Steam Cooling Performance in a Ribbed Passage

This study numerically examines the effect of actual gas turbine operating conditions on heat transfer characteristics in a ribbed passage with mist/steam cooling. A 60° ribbed passage with aspect ratio of 1/1 was investigated at Reynolds number of 300,000, and steam cooling was used to provide a contrast. Three main factors were considered: coolant temperature, operating pressure, and wall heat flux density. The heat transfer enhancement mechanism of mist/steam cooling was explored, and the results showed that the heat transfer performance of mist/steam cooling was superior to steam cooling. When the coolant temperature varied from 300 to 500 °C, the average Nusselt number of mist/steam cooling decreased by 26.6%, and the heat transfer enhancement ratio dropped from 15% to 10%. As operating pressure increased, the heat transfer performance factor of mist/steam firstly increased and then decreased. At an operating pressure of 1.5 MPa, the heat transfer achieved its optimal performance, and the heat transfer enhancement ratio achieved its maximum value of 15.9%. Larger wall heat flux density provided less heat transfer enhancement. When the heat flux density increased from 100,000 to 300,000 W·m−2, the average Nusselt number of mist/steam cooling decreased by 13.8%, while the heat transfer enhancement ratio decreased from 25.3% to 12.6%

A Numerical Investigation of Frost Growth on Cold Surfaces Based on the Lattice Boltzmann Method

A numerical investigation of frost growth on a cold flat surface was presented based on two-dimensional Lattice Boltzmann model. This model has been validated to have less prediction error by past experiments. According to the results, it is shown that average frost density appears different at an increasing rate at different frosting stages. In addition, cold surface temperature has great influence on frost growth parameters such as frost crystal deposition mass, frost deposition rate, and frost crystal volume fraction. It was found that the frost crystal deposition mass, frost crystal volume, and the deposition rate first increase rapidly, then gradually slow down, finally remaining unchanged while the cold surface temperature decreases. The further away from the cold surface, the more sparser the frost layer structure becomes due to the smaller frost crystal volume fraction