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Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) is a novel, highly infectious RNA virus that belongs to the coronavirus family. Replication of the viral genome is a fundamental step in the virus life cycle and SARS‐CoV‐2 non‐structural protein 9 (Nsp9) is shown to be essential for virus replication through its ability to bind RNA in the closely related SARS‐CoV‐1 strain. Two recent studies revealing the three‐dimensional structure of Nsp9 from SARS‐CoV‐2 have demonstrated a high degree of similarity between Nsp9 proteins within the coronavirus family. However, the binding affinity to RNA is very low which, until now, has prevented the determination of the structural details of this interaction. In this study, we have utilized nuclear magnetic resonance spectroscopy (NMR) in combination with surface biolayer interferometry (BLI) to reveal a distinct binding interface for both ssDNA and RNA that is different to the one proposed in the recently solved SARS‐CoV‐2 replication and transcription complex (RTC) structure. Based on these data, we have proposed a structural model of a Nsp9‐RNA complex, shedding light on the molecular details of these important interactions. [ABSTRACT FROM AUTHOR]

Near-Infrared Spectroscopy (NIRS) has been used to measure muscle mitochondrial capacity (mVO2max) as the recovery rate constant of muscle metabolism after exercise. The current method requires as many as 50 short ischemic occlusions to generate 2 recovery rate constants., Purpose: To determine the effectiveness of using a 6-occlusion protocol (Mito6) versus one with 22 occlusions (Mito22) to measure muscle mitochondrial capacity., Method: In two independent data sets (bicep n=7, forearm A n=23), recovery curves were analyzed independently using both the Mito6 and Mito22 analyses. A third data set (Forearm B, n=16) was generated on forearm muscles of healthy subjects using four Mito6 tests performed in succession. Recovery rate constants were generated using a MATLAB routine., Results: When calculated from the same data set, the recovery rate constants were not significantly different between the Mito22 and Mito6 analyses for the bicep (1.43+0.33min-1, 1.43+0.35min-1, p=0.81) and the forearm A (1.97+0.40min-1, 1.97+0.43min-1, p=0.90). The correlation between Mito22 and Mito6 recovery rate constants was y=1.07x-0.09, R2=0.90 for the bicep data and 1.00x+0.01, R2=0.85 for the forearm A data. When performing the four Mito6 tests in the Forearm B study; recovery rate constants were not different between tests (1.50±0.51 min-1, 1.42±0.54 min-1, 1.26±0.41 min-1, 1.29±0.47 min-1, P>0.05)., Conclusions: Muscle mitochondrial capacity was not different between the Mito6 analysis and the longer Mito22 analysis. The Mito6 protocol was considered more practical as it used fewer ischemic occlusion periods, and multiple tests could be performed in succession in less time. There were no order effects for the rate constants of four repeated Mito6 tests of mitochondrial capacity, supporting the use of multiple tests to improve accuracy.