Heme from Alzheimer's brain inhibits muscarinic receptor binding via thiyl radical generation.

An endogenous inhibitor (< 3500 Da) of antagonist binding to the muscarinic acetylcholine receptor (mAChR) has been reported to be elevated 3-fold in Alzheimer's disease (AD) brain. This endogenous inhibitor was found to require the presence of reducing agents such as reduced glutathione (GSH) for optimal activity. In the presence of GSH, the inhibitor was shown to generate thiyl radicals which irreversibly inhibited the mAChR. We now report that the inhibitor contains free heme, a well-established source of oxidative stress capable of generating free radicals and causing neurotoxicity. While FeSO4, microperoxidase and hemin all inhibited antagonist binding to the mAChR, only hemin shared the inhibitor's requirement for GSH. Both the free radical scavengers Trolox and Mn2+, and the metal chelator, EDTA, blocked the activity of the endogenous AD inhibitor and of hemin. Heme oxygenase-1 (HO-1) markedly reduced the activity of both the endogenous AD inhibitor and hemin, indicating that the endogenous inhibitor contains heme. Mass spectrometric analysis confirmed the presence of free heme and heme fragments in fractions of the endogenous AD inhibitor. The antioxidants estrogen, vitamin E and vitamin C all protected the mAChR from irreversible inhibition by the endogenous inhibitor or hemin. These antioxidants may function to protect the integrity of the mAChR in vivo and may have therapeutic potential in AD where free heme could be a source of oxidative stress.

Endogenous Alzheimer's brain factor and oxidized glutathione inhibit antagonist binding to the muscarinic receptor.

An endogenous inhibitor (< 3,500 Da) of antagonist binding to the muscarinic acetylcholine receptor has been extracted from Alzheimer's disease (AD) brain with trifluoracetic acid. Oxidized glutathione, (GSSG) has also been found to inhibit antagonist binding to the receptor. However, in its reduced form, glutathione (GSH) like other reducing agents, markedly enhances the inhibitory effect of both GSSG and the endogenous AD inhibitor. EDTA and the free radical scavengers Mn(2+) and Trolox, a vitamin E analog, block the action of the endogenous AD inhibitor but not of GSSG in the presence of GSH. Further, while GSSG inhibition is reversible, the action of the endogenous AD inhibitor is irreversible, consistent with a free radical mechanism. The enhancement of endogenous AD inhibitor activity by GSH suggested that GSH may be involved in formation of the free radical generated by the inhibitor. The glutathione thiyl radical is shown to inhibit antagonist binding to the receptor and is, therefore, a good candidate for the free radical formed by the endogenous AD inhibitor. The ability of Trolox to block the reduction in muscarinic receptor binding caused by the endogenous AD inhibitor is encouraging and suggests that free radical scavengers, such as vitamin E, may have a potential therapeutic role in AD by protecting the integrity of the muscarinic receptor.

Medical coverage for multievent sports competition: a comprehensive analysis of injuries in the 1994 Star of the North Summer Games.

OBJECTIVE: To determine the incidence and type of requests for medical assistance and the treatment required in 21 sports at the 1994 Star of the North Summer Games and to develop a plan, based on these findings, for efficient allocation of medical supplies and staff. DESIGN: We reviewed our experience with a large multisport amateur athletic event and analyzed the pattern of injuries for participants in the various sports. MATERIAL AND METHODS: Injury evaluation forms and medical supply kits were given to the health-care volunteers, who received preliminary instructions on classifications and definitions of injuries and on appropriate completion of the forms. When a medical contact occurred, an evaluation form was completed. The data from these forms were subsequently compiled and analyzed. RESULTS: Of 6,243 athletes who participated in the 1994 Star of the North Summer Games, 55 (0.88%) received medical attention (2 officials also required medical assistance). The sports with the greatest number of medical contacts were soccer and track and field--31 and 16 contacts for medical assistance, respectively. The lower extremities were the anatomic site most frequently involved in injury (62% of the medical contacts). The most common types of injury were contusions, strains, and sprains. CONCLUSION: Because few injuries were sustained and most were of minor severity, basic medical supplies (such as ice bags and compression wraps) were sufficient. For locations with a scarcity of medical personnel, a triage system can be established in which athletic trainers or registered nurses manage the initial assessments and refer cases, as needed, to physicians for specialized care. An effective communication system can also considerably reduce volunteer hours.