No abstract

Nicotinamide adenine dinucleotide (NAD+) is an essential metabolite utilized as a redox cofactor and enzyme substrate in numerous cellular processes. Elevated NAD+ levels have been observed in red blood cells infected with the malaria parasite Plasmodium falciparum, but little is known regarding how the parasite generates NAD+. Here, we employed a mass spectrometry-based metabolomic approach to confirm that P. falciparum lacks the ability to synthesize NAD+ de novo and is reliant on the uptake of exogenous niacin. We characterized several enzymes in the NAD+ pathway and demonstrate cytoplasmic localization for all except the parasite nicotinamidase, which concentrates in the nucleus. One of these enzymes, the P. falciparum nicotinate mononucleotide adenylyltransferase (PfNMNAT), is essential for NAD+ metabolism and is highly diverged from the human homolog, but genetically similar to bacterial NMNATs. Our results demonstrate the enzymatic activity of PfNMNAT in vitro and demonstrate its ability to genetically complement the closely related Escherichia coli NMNAT. Due to the similarity of PfNMNAT to the bacterial enzyme, we tested a panel of previously identified bacterial NMNAT inhibitors and synthesized and screened twenty new derivatives, which demonstrate a range of potency against live parasite culture. These results highlight the importance of the parasite NAD+ metabolic pathway and provide both novel therapeutic targets and promising lead antimalarial compounds.

4 Laryngoscope, 127:900-906, 2017.

STRUCTURED ABSTRACT OBJECTIVES The literature increasingly demonstrates the importance of gait speed in the frailty assessment of patients 60 years and older. Conventional gait speed measurement, however, may be contraindicated in settings such as trauma where the patient is temporarily immobilized. We devised a Walking Speed Questionnaire (WSQ) to allow assessment of pre-injury baseline gait speed, in meters per second, in a self-reported manner, to overcome the inability to directly test the patients’ walking speed. DESIGN Four questions comprise the WSQ, and were derived using previously published questionnaires and expert opinion of six physician-researchers. SETTING Four ambulatory clinics. PARTICIPANTS Ambulating individuals aged 60 to 95 (mean age 73.2 ± 8.1, 86.1% female, N = 101). INTERVENTION Participants completed the WSQ and underwent gait speed measurement for comparison. ANALYSIS WSQ score correlation to true gait speed, receiver operating characteristics, and validation statistics were performed. RESULTS All four questions of the WSQ independently predicted true gait speed significantly (P<0.001). The WSQ sufficiently predicted true gait speed with r = 0.696 and ρ = 0.717. CONCLUSION The WSQ is an effective tool for assessing baseline walking speed in patients 60 years and older in a self-reported manner. It permits gait screening in health care environments where conventional gait speed testing is contraindicated due to temporary immobilization, and may be used to provide baseline targets for goal-oriented post-trauma care. Given its ability to capture gait speed in patients who are unable to ambulate, it may open doors for frailty research in previously unattainable populations.