In the accompanying paper [Matsubara, M., et al. (2003) Biochemistry 42, 4993-5002], we have partially purified and characterized rat 5-formyluracil (fU)-DNA glycosylase (FDG). Several lines of evidence have indicated that FDG is a rat homologue of single-strand-selective monofunctional uracil-DNA glycosylase (SMUG1). We report here that rat and human SMUG1 (rSMUG1 and hSMUG1) expressed from the corresponding cDNAs indeed excise fU in single-stranded (ss) and double-stranded (ds) DNA. The enzymes also excised uracil (U) and uracil derivatives bearing an oxidized group at C5 [5-hydroxyuracil (hoU) and 5-hydroxymethyluracil (hmU)] in ssDNA and dsDNA but not analogous cytosine derivatives (5-hydroxycytosine and 5-formylcytosine) and other oxidized damage. The damage specificity and the salt concentration dependence of rSMUG1 (and hSMUG1) agreed well with those of FDG, confirming that FDG is rSMUG1. Consistent with the damage specificity above, hSMUG1 removed damaged bases from Fenton-oxidized calf thymus DNA, generating abasic sites. The amount of resulting abasic sites was about 10% of that generated by endonuclease III or 8-oxoguanine glycosylase in the same substrate. The HeLa cell extract and hSMUG1 exhibited a similar damage preference (hoU.G > hmU.A, fU.A), and the activities for fU, hmU, and hoU in the cell extract were effectively neutralized with hSMUG1 antibodies. These data indicate a dual role of hSMUG1 as a backup enzyme for UNG and a primary repair enzyme for a subset of oxidized pyrimidines such as fU, hmU, and hoU.

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We investigated the effects of increasing CO2 ventilatory drive on the coordination of respiration and reflex swallowing elicited by continuous infusion of distilled water into the pharynx (2.5 ml/min) in 11 normal subjects. Ventilation was monitored using a pneumotachograph and swallowing was recorded by submental electromyogram. The CO2 ventilatory drive was increased by addition of external dead space, while ventilation, the frequency of swallows, and the timing of swallows in relation to the phases of the respiratory cycle were measured at steady-state conditions. We found that the CO2 ventilatory response is not influenced by continuous reflex swallowing but that hypercapnia influences the timing and frequency of these swallows. Signs of aspiration were never observed during continuous infusion of water at eucapnia, but seven of 11 subjects showed laryngeal irritation and/or pending aspiration during hypercapnia, and the incidence of laryngeal irritation was higher the greater the PCO2. Detailed analysis of laryngeal irritations consisting of single coughs in seven subjects revealed that the majority of laryngeal irritations occurred when swallows coincided with expiratory-inspiratory transition or when swallows coincided with inspiration, whereas laryngeal irritation after an expiratory swallow was never observed. These results suggest that the automatic respiratory control system is not influenced by continuous swallowing but that the coordination of swallowing and respiration may be compromised during hypercapnia.

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