Interobserver and Intraobserver Variability in the CT Assessment of COVID-19 Based on RSNA Consensus Classification Categories.

PURPOSE: To assess the interobserver and intraobserver agreement of fellowship trained chest radiologists, nonchest fellowship-trained radiologists, and fifth-year radiology residents for COVID-19-related imaging findings based on the consensus statement released by the Radiological Society of North America (RSNA). METHODS: A survey of 70 chest CTs of polymerase chain reaction (PCR)-confirmed COVID-19 positive and COVID-19 negative patients was distributed to three groups of participating radiologists: five fellowship-trained chest radiologists, five nonchest fellowship-trained radiologists, and five fifth-year radiology residents. The survey asked participants to broadly classify the findings of each chest CT into one of the four RSNA COVID-19 imaging categories, then select which imaging features led to their categorization. A 1-week washout period followed by a second survey comprised of randomly selected exams from the initial survey was given to the participating radiologists. RESULTS: There was moderate overall interobserver agreement in each group (κ coefficient range 0.45-0.52 ± 0.02). There was substantial overall intraobserver agreement across the chest and nonchest groups (κ coefficient range 0.61-0.67 ± 0.06) and moderate overall intraobserver agreement within the resident group (κ coefficient 0.58 ± 0.06). For the image features that led to categorization, there were varied levels of agreement in the interobserver and intraobserver components that ranged from fair to perfect kappa values. When assessing agreement with PCR-confirmed COVID status as the key, we observed moderate overall agreement within each group. CONCLUSION: Our results support the reliability of the RSNA consensus classification system for COVID-19-related image findings.

Glutathionylation of α-ketoglutarate dehydrogenase: the chemical nature and relative susceptibility of the cofactor lipoic acid to modification.

α-Ketoglutarate dehydrogenase (KGDH) is reversibly inhibited when rat heart mitochondria are exposed to hydrogen peroxide (H2O2). H2O2-induced inhibition occurs through the formation of a mixed disulfide between a protein sulfhydryl and glutathione. Upon consumption of H2O2, glutaredoxin can rapidly remove glutathione, resulting in regeneration of enzyme activity. KGDH is a key regulatory site within the Krebs cycle. Glutathionylation of the enzyme may therefore represent an important means to control mitochondrial function in response to oxidative stress. We have previously provided indirect evidence that glutathionylation occurs on lipoic acid, a cofactor covalently bound to the E2 subunit of KGDH. However, lipoic acid contains two vicinal sulfhydryls and rapid disulfide exchange might be predicted to preclude stable glutathionylation. The current study sought conclusive identification of the site and chemistry of KGDH glutathionylation and factors that control the degree and rate of enzyme inhibition. We present evidence that, upon reaction of free lipoic acid with oxidized glutathione in solution, disulfide exchange occurs rapidly, producing oxidized lipoic acid and reduced glutathione. This prevents the stable formation of a glutathione-lipoic acid adduct. Nevertheless, 1:1 lipoic acid-glutathione adducts are formed on KGDH because the second sulfhydryl on lipoic acid is unable to participate in disulfide exchange in the enzyme's native conformation. The maximum degree of KGDH inhibition that can be achieved by treatment of mitochondria with H2O2 is 50%. Results indicate that this is not due to glutathionylation of a subpopulation of the enzyme but, rather, the unique susceptibility of lipoic acid on a subset of E2 subunits within each enzyme complex. Calcium enhances the rate of glutathionylation by increasing the half-life of reduced lipoic acid during enzyme catalysis. This does not, however, alter the maximal level of inhibition, providing further evidence that specific lipoic acid residues within the E2 complex are susceptible to glutathionylation. These findings offer chemical information necessary for the identification of mechanisms and physiological implications of KGDH glutathionylation.