An activity transition from NADH dehydrogenase to NADH oxidase during protein denaturation.

A decrease in the specific activity of an enzyme is commonly observed when the enzyme is inappropriately handled or is stored over an extended period. Here, we reported a functional transition of an FMN-bound diaphorase (FMN-DI) that happened during the long-term storage process. It was found that FMN-DI did not simply lose its ß-nicotinamide adenine diphosphate (NADH) dehydrogenase activity after a long-time storage, but obtained a new enzyme activity of NADH oxidase. Further mechanistic studies suggested that the alteration of the binding strength of an FMN cofactor with a DI protein could be responsible for this functional switch of the enzyme.

A Hidden Transhydrogen Activity of a FMN-Bound Diaphorase under Anaerobic Conditions.

BACKGROUND: Redox cofactors of NADH/NADPH participate in many cellular metabolic pathways for facilitating the electron transfer from one molecule to another in redox reactions. Transhydrogenase plays an important role in linking catabolism and anabolism, regulating the ratio of NADH/NADPH in cells. The cytoplasmic transhydrogenases could be useful to engineer synthetic biochemical pathways for the production of high-value chemicals and biofuels. METHODOLOGY/PRINCIPAL FINDINGS: A transhydrogenase activity was discovered for a FMN-bound diaphorase (DI) from Geobacillus stearothermophilus under anaerobic conditions. The DI-catalyzed hydride exchange were monitored and characterized between a NAD(P)H and a thio-modified NAD+ analogue. This new function of DI was demonstrated to transfer a hydride from NADPH to NAD+ that was consumed by NAD-specific lactate dehydrogenase and malic dehydrogenase. CONCLUSIONS/SIGNIFICANCE: We discover a novel transhydrogenase activity of a FMN-DI by stabilizing the reduced state of FMNH2 under anaerobic conditions. FMN-DI was demonstrated to catalyze the hydride transfer between NADPH and NAD+. In the future, it may be possible to incorporate this FMN-DI into synthetic enzymatic pathways for balancing NADH generation and NADPH consumption for anaerobic production of biofuels and biochemicals.

Whole-body 18F-FDG PET/CT: the need for a standardized field of view--a referring-physician aid.

UNLABELLED: PET/CT fusion of anatomic and functional imaging modalities is in evolution, with rapid clinical dissemination. The imaged field of view (FOV) selected for whole-body PET/CT protocols is not standardized and varies by institution. Misuse of the term whole body, as well as the pressure to increase the number of daily studies by reducing scanning time, contributes to the lack of standardization. The purpose of this study was to evaluate variations in the FOV and arm positioning selected for whole-body PET/CT protocols at private, as well as academic, PET centers. METHODS: Two hundred consecutive whole-body (18)F-FDG PET/CT studies were retrospectively reviewed for FOV: 50 studies from a private stationary site, 50 studies from 2 separate private mobile sites (25 consecutive studies from each), and 100 studies from a stationary university site: 50 before and 50 after implementation of a true whole-body protocol covering the top of the head through the bottom of the feet. Data were categorized into 5 different anatomic scan lengths: base of skull to upper thigh, base of skull to mid thigh, top of head to upper thigh, top of head to mid thigh, and true whole-body. Studies were further categorized into 2 patient arm positions: up and down. RESULTS: The private stationary and mobile sites had only 2 categories of anatomic scan lengths identified: base of skull to mid thigh, and top of head to upper thigh. At the university site, before implementation of a true whole-body protocol, the 5 different anatomic scan lengths were identified; after implementation, only the true whole-body scan length was identified. Patients' arms in the private stationary sites were down 100% of the time. At the private mobile sites, patients' arms were up 72% of the time and down 28% of the time. At the university site, patients' arms were up 54% of the time and down 46% of the time. The same site, after implementation of a true whole-body protocol, had patients' arms up 58% of the time and down 42% of the time. Overall, patients' arms were up 46% of the time and down 54% of the time. CONCLUSION: The continued use of the term whole body is misleading because frequently it may not include the brain, skull, or significant portions of the upper and lower extremities. PET/CT anatomic scan length varied not only from one site to the next but also within individual sites. The Centers for Medicare and Medicaid Services have different current procedural terminology codes distinguishing between base of skull to upper thigh and true whole-body covering the top of the skull to the bottom of the feet, thus underscoring the need to standardize the terminology used in describing PET/CT scan length.