╨╧рб▒с>■   ■                                                                                                                                                                                                                                                                                                                                                                                                                                   ье┴[@ °┐sbjbj44 .ViVik      ИrrrrrrrЦ^;^;^;^;$В<$Ц,гЎ▓=╪КQКQКQКQ#Tb Еa▄afpлвнвнвнвнвнвнв$"дRtж·╤вr╤hT"#T╤h╤h╤вrrКQКQwцвWЮWЮWЮ╤hЁ"rКQrКQлвWЮ╤hлвWЮDWЮЫЮrrЫЮКQж= `#яd"╔┬^;┴ЛZЫЮ╫Ю╘№в0,гЫЮnзЬ@nзЫЮЖD╩╠rrrrЫЮnзrпЮ(╤h╤hWЮ╤h╤h╤h╤h╤h╤в╤вЦЦфz ф[Ь№ЦЦz Journal of the American Academy of Dermatology Volume 43 Х Number 2 Х August 2000 Copyright й 2000 American Academy of Dermatology, Inc.  INCLUDEPICTURE "http://home.mdconsult.com/images/blank.gif" \* MERGEFORMATINET  INCLUDEPICTURE "http://home.mdconsult.com/images/blank.gif" \* MERGEFORMATINET  INCLUDEPICTURE "http://home.mdconsult.com/images/blank.gif" \* MERGEFORMATINET THERAPY  Pharmacology of botulinum toxin Wilber Huang MDa Jill A. Foster MDb Arlene S. Rogachefsky MDa  From the Departments of Dermatologya and Ophthalmology,b Cleveland Clinic Foundation.  Reprint requests: Wilber Huang, MD, 31701 Isle Vista Dr, Laguna Niguel, CA 92677. Copyright й 2000 by the American Academy of Dermatology, Inc. 0190-9622/2000/$12.00 + 0аа16/1/105567 Cleveland, Ohio Background: Botulinum toxin has a well-defined role among dermatologists for the treatment of facial wrinkling, brow position, and palmar and axillary hyperhidrosis. Objective: The purpose of this study is to educate dermatologists on the pharmacology of botulinum toxin. Methods: A retrospective review of the literature on botulinum toxin from 1962 to the present was conducted. We examined the clinical applications of botulinum toxin, cholinergic neuromuscular transmission, the toxin's structure and molecular actions, drug and disease interactions at the neuromuscular junction, toxin assays, determinants of clinical response, and adverse side effects. Results: Botulinum toxin blocks the release of acetylcholine from the presynaptic terminal of the neuromuscular junction. Several drugs and diseases interfere with the neuromuscular junction and the effects of botulinum toxin. The mouse bioassay, the most sensitive and specific measurement of toxin activity, is the gold standard for botulinum toxin detection and standardization. The major determinants of clinical response to treatment are the toxin preparation, individual patient's anatomy, dose and response relationships, length of toxin storage after reconstitution, and immunogenicity. To minimize potential antibody resistance, one should use the smallest effective dose, utilize treatment intervals of more than 3 months, and avoid booster injections. Uncommon adverse effects include ptosis, ectropion, diplopia, bruising, eyelid drooping, hematoma formation, and temporary headaches. Conclusion: Botulinum toxin is a safe and effective treatment. Knowledge of the pharmacologic basis of therapy will be useful for standardizing techniques and achieving consistent therapeutic results in the future. (J Am Acad Dermatol 2000;43:249-59.) Background Neurotoxins produced by the gram-positive, anaerobic Clostridium botulinum are the most potent toxins known to mankind and are the causative agents of botulism. Botulinum toxin (BTX) acts by blocking the release of acetylcholine from the presynaptic terminal of the neuromuscular junction. Seven distinct antigenic botulinum toxins (BTX-A, B, C, D, E, F, and G) produced by different strains of Clostridium botulinum have been described.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567001#R105567001" 1] The human nervous system is susceptible to 5 toxin serotypes (BTX-A, B, E, F, G) and unaffected by 2 (BTX-C, D).[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567002#R105567002" 2] There are 3 forms of human botulism: food-borne, infantile, and wound.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567003#R105567003" 3] Classic or food-borne botulism results from the ingestion of food containing preformed neurotoxin types A, B, or E.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567004#R105567004" 4] It is characterized by a symmetric, descending flaccid paralysis of motor and autonomic nerves, usually beginning with the cranial nerves. Blurred vision, dysphagia, and dysarthria are common initial symptoms and in the severest cases can lead to respiratory arrest and death.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567005#R105567005" 5] Infant botulism is caused by ingestion of spores and production of toxin in the infant's intestine. Wound botulism arises as a consequence of toxin produced in wounds contaminated with the organism.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567006#R105567006" 6] The potential medical applications of BTX-A were first recognized by Scott[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567007#R105567007" 7] in the early 1980s when he used local injection of minute doses to selectively inactivate muscle spasticity in strabismus. BTX-A was found to be a safe and effective therapy without significant local or systemic side effects. This success and a series of clinical studies[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567008#R105567008" 8] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567011#R105567011" 11] led to Food and Drug Administration (FDA) approval in 1989 for ophthalmologic and neurologic use in strabismus, blepharospasm, and hemifacial spasm. BTX-A has since been used in other conditions related to excessive muscle activity such as cervical dystonia,[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567012#R105567012" 12] spasmodic torticollis, [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567013#R105567013" 13] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567014#R105567014" 14] and achalasia[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567015#R105567015" 15] (Table I). Table I. Clinical use of BotoxDermatologyOphthalmologyNeurologyGlabellar frown linesStrabismusHemifacial spasmCrow's feetBlepharospasmFacial asymmetryForehead linesNystagmusOromandibular dystoniaPerioral linesCervical dystoniaPlatysmal bandsSpasmodic torticollisBrow ptosisAchalasiaPalmar hyperhidrosisGustatory sweatingAxillary hyperhidrosisSynkinesiaHyperlacrimation BTX use in the field of dermatology (Table I) began in the early 1990s when improvement in facial wrinkling was observed. While treating hemifacial spasm patients with BTX-A, Borodic, Cheney, and McKenna[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567016#R105567016" 16] reported a concomitant unilateral decrease in facial wrinkling. Carruthers and Carruthers [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567017#R105567017" 17] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567018#R105567018" 18] treated patients with blepharospasm and noticed an improvement in glabellar frown lines. These initial findings and subsequent clinical studies popularized BTX-A as a safe and effective treatment for hyperfunctional glabellar frown lines, crow's feet, and forehead lines.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567017#R105567017" 17] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567023#R105567023" 23] More recently, BTX-A has been used to treat platysmal bands,[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567024#R105567024" 24] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567025#R105567025" 25] brow position,[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567019#R105567019" 19] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567026#R105567026" 26] and palmar[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567027#R105567027" 27] and axillary hyperhidrosis.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567028#R105567028" 28] Neuromuscular transmission Cholinergic neurotransmission involves 6 steps: synthesis, storage, release, binding, degradation, and recycling of acetylcholine[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567029#R105567029" 29] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567030#R105567030" 30] (Fig 1).  HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/I35.fig" \l "top" Fig. 1. Cholinergic neurotransmission and botulinum toxin's mechanism of action. Choline is first transported from the extracellular fluid into the cholinergic neuron's cytoplasm by a carrier system that cotransports sodium. Choline reacts enzymatically with acetylcholine CoA to form acetylcholine, which is then transported into synaptic vesicles where it is stored in granules. When an action potential arrives at a nerve ending, voltage-sensitive calcium channels in the presynaptic membrane open causing an increase in the concentration of intracellular calcium. Elevated calcium levels promote the docking and fusion of synaptic vesicles with the cell membrane via a complex mechanism involving protein isoforms, culminating in the release of acetylcholine. Acetylcholine then diffuses across the synaptic space and binds to postsynaptic nicotinic receptors on the muscle fiber. This binding activates a second messenger system that results in muscle contraction. Acetylcholine is rapidly cleaved into choline and acetate by acetylcholinesterase. Choline may be recycled by a high-affinity transport system that pulls the molecule back into the neuron. Toxin structure Botulinum neurotoxins are produced as inactive polypeptides of 150 kd, which are cleaved by trypsinlike bacterial protease to generate the di-chain active form of the toxin. The proportion of single to di-chain toxin is dependent on the toxin's serotype and whether or not the bacterial strain expresses the appropriate protease.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567031#R105567031" 31] The 100-kd heavy (H) chains and the 50-kd light (L) chains are linked together by heat-labile disulfide bonds and noncovalent forces.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567032#R105567032" 32] The H and L chains dissociate with heat and boiling, which inactivates the toxin because neurotoxicity requires both H and L chains.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567033#R105567033" 33] Molecular actions All serotypes act on the peripheral nervous system where they inhibit release of acetylcholine from the presynaptic terminal of the neuromuscular junction. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567034#R105567034" 34] Toxins may bind to nerve terminals at autonomic cholinergic ganglia with autonomic effects, but only in very large doses. It is unlikely that therapeutic doses are associated with any significant autonomic adverse reactions.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567033#R105567033" 33] There are three steps involved in neurotoxicity[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567035#R105567035" 35] (Fig 1). Binding The first step is the irreversible binding of BTX to presynaptic cholinergic receptors via the H chain's 50-kd carboxy-terminal.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567036#R105567036" 36] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567038#R105567038" 38] The associated binding sites have not been clearly characterized. Previous studies have suggested that distinct receptors exist for different BTX serotypes.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567036#R105567036" 36] This view has been challenged by the isolation of a highly conserved synaptic vesicle protein, synaptotagmin, which binds to BTX-A, BTX-B, and BTX-E.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567039#R105567039" 39] Internalization The second step involves internalization of the neurotoxin through a receptor-mediated endocytosis.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567040#R105567040" 40] This process is independent of calcium and partially dependent on nerve stimulation. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567041#R105567041" 41] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567042#R105567042" 42] After internalization, the disulfide bond is cleaved by an unknown mechanism. The H chain's 50-kd amino-terminal is associated with ionic channel formation and translocation of the L-chain from the endosome into the neuronal cytoplasm.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567043#R105567043" 43] Neuromuscular blockade The third step is neuromuscular blockade. Within the synapse, protein isoforms form a complex platform necessary for the docking, fusion, and release of acetylcholine vesicles through the cell membrane.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567044#R105567044" 44] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567045#R105567045" 45] These protein isoforms are vesicle associated membrane protein (VAMP, also known as synaptobrevin), synaptosomal associated protein (SNAP-25), and syntaxin. The L- chain of each toxin, which contains a highly specific zinc-endopeptidase with proteolytic activity concentrated at its aminoterminal, cleaves a single protein isoform at a single site[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567033#R105567033" 33] (Table II). Table II. Target substrates of botulinum toxinToxin typeSubstrateReference No(s).BTX-ASNAP-25[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567122#R105567122" 122] 122BTX-BVAMP/synaptobrevin[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567123#R105567123" 123] 123BTX-CSNAP-25 and Syntaxin[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567124#R105567124" 124] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567125#R105567125" 125] 124,125BTX-DVAMP/synaptobrevin[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567122#R105567122" 122] 122BTX-ESNAP-25[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567122#R105567122" 122] 122BTX-FVAMP/synaptobrevin[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567126#R105567126" 126] 126BTX-GVAMP-synaptobrevin[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567127#R105567127" 127] 127 The only exception is BTX-C, which cleaves two proteins. Drug and disease interaction Several drugs act on the neuromuscular junction and interfere with the effect of BTX (Table III). Table III. Drug and disease interactionsDrug/diseaseMechanismsPresynaptic nerve terminalаааAminoglycosides (kanamycin, streptomycin, gentamicin)Calcium channel blockadeаааAminoquinolines (chloroquine, hydroxychloroquine)Inhibit botulinum toxin binding to synaptotagmin or lysosomal processing of toxinаааCyclosporineCalcium channel blockadeаааLambert-Eaton syndromeCross-reacting, tumor antigen antibodies against calcium channelsPostsynaptic nerve terminalаааMyasthenia gravisAutoantibodies to the nicotinic acetylcholine receptorаааD-PenicillamineTriggers the formation of nicotinic acetylcholine receptor antibodiesаааTubocurarine, pancuronium, gallaminePostsynaptic acetylcholine antagonist blockersаааSuccinylcholinePostsynaptic acetylcholine agonist blocker BTX may be potentiated by aminoglycoside antibiotics.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567046#R105567046" 46] Large doses of aminoglycosides such as kanamycin, streptomycin, and gentamicin can prevent the release of acetylcholine from nerve endings and produce a botulism-like clinical syndrome. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567047#R105567047" 47] This effect may be related to calcium channel blockade.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567048#R105567048" 48] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567049#R105567049" 49] Symptoms rapidly abate as the offending drug is eliminated from the body. Aminoquinolines (chloroquine and hydroxychloroquine) antagonize the onset of paralysis from BTX by acting either at the cell membrane to inhibit toxin binding or internalization, or in the cell interior to inhibit lysosomal processing of toxin.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567050#R105567050" 50] Cyclosporine has been reported to cause neuromuscular blockade characterized by muscle weakness and ventilatory failure.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567051#R105567051" 51] The precise mechanism of action is unknown but may be the result of anti-inflammatory or immunosuppressive effects on the muscle or presynaptic calcium channel blockade.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567052#R105567052" 52] D-Penicillamine can trigger the formation of acetylcholine receptor antibodies in immunologically predisposed individuals.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567053#R105567053" 53] A small percentage of patients with rheumatoid arthritis who receive D-penicillamine develop acetylcholine receptor antibodies[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567054#R105567054" 54] and symptoms of myasthenia gravis.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567055#R105567055" 55] Both the symptoms and antibodies remit within a few months after drug cessation. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567056#R105567056" 56] The antibody repertoire in the sera of patients with myasthenia gravis and D-penicillamine-induced myasthenia gravis resembles each other.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567057#R105567057" 57] A few nondermatologic drugs block cholinergic transmission between motor nerve endings and the postsynaptic nicotinic receptors on the neuromuscular end-plate. They act either as competitive, antagonist blockers (tubocurarine, pancuronium, and gallamine) or as agonist blockers (succinylcholine). The antagonist blockers compete with acetylcholine for receptor binding sites. A sufficient amount of drug will block the acetylcholine receptors, thereby preventing muscle depolarization. The mechanism of agonist blockers resembles the depolarizing action of acetylcholine. However, the muscle repolarization from succinylcholine is much slower than acetycholine, causing a delay in muscle contraction.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567058#R105567058" 58] BTX use is contraindicated in patients with disorders of neuromuscular transmission (eg, Lambert-Eaton syndrome and myasthenia gravis). In Lambert-Eaton syndrome, antibodies directed against tumor antigens cross-react with voltage-gated calcium channels involved in acetylcholine release, leading to a disturbance of neuromuscular transmission. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567059#R105567059" 59] Weakness in myasthenia gravis is caused by an antibody-induced internalization and degradation of acetylcholine receptors.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567060#R105567060" 60] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567061#R105567061" 61] Assays and pharmacologic actions Muscle assays Several assays are invaluable in elucidating the pharmacologic action of BTX. The mouse phrenic nerve diaphragm model established the multi-step hypothesis of BTX.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567062#R105567062" 62] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567063#R105567063" 63] Toxin radiolabeling coupled with histologic analysis delineated the relationship among toxins, receptor sites, and motor end plates. It demonstrated that nerves of actively contracting muscles bind BTX more rapidly[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567041#R105567041" 41] at an average of 32 to 64 minutes.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567048#R105567048" 48] This phenomenon supports the directive for patients to exercise their injected muscles for up to an hour after BTX injection. The mouse hypoglossal nerve assays showed that unlike doxorubicin, which causes toxicity to the neuron cell body by retrograde transport,[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567064#R105567064" 64] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567065#R105567065" 65] BTX does not induce motorneuron death.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567066#R105567066" 66] Rather, it causes a chemical denervation to the neuromuscular junction.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567067#R105567067" 67] Studies of muscle fibers from human blepharospasm patients demonstrated that denervation is reversible. Using acetylcholinesterase stain to assess denervation of striated muscle after injection of BTX, Borodic et al[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567068#R105567068" 68] observed that denervation is accompanied by spreading of the acetylcholinesterase activity to cover most of the exposed sarcolemma. After 4 to 5 months, the distribution of acetylcholinesterase activity reverts to its normal pattern. Recovery from denervation occurs by neurogenesis, with the formation of axonal sprouts within 10 days and new motor end plates.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567069#R105567069" 69] A reconnection between nerve terminals and muscle motor end plates is therefore established. The rabbit longissimus dorsi muscle assay showed that muscle atrophy is also reversible. Localized muscle atrophy is seen within 2 weeks of toxin injection, continues for about 4 weeks, and then reverses.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567070#R105567070" 70] Similar reversible denervation atrophy has been seen in the orbicularis oculi muscles of bleph-arospasm patients treated with BTX-A.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567068#R105567068" 68] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567071#R105567071" 71] Clinical recovery of function requires about 3 to 6 months, at which time the muscle returns to about 70% to 80% of its original bulk.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567033#R105567033" 33] Sprouting and remodeling may continue for up to 3 years.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567072#R105567072" 72] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567073#R105567073" 73] Bioassay The mouse biologic assay is currently the only accepted quantitative method for the detection of Clostridium toxins in culture, serum, and food samples,[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567074#R105567074" 74] and antitoxin standardization.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567075#R105567075" 75] It is the most sensitive and specific measurement of BTX activity. Factors affecting the assay include sex, strain, species, and the route of administration (intraperitoneal vs intramuscular).[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567076#R105567076" 76] One mouse unit (MU) is defined as the median intraperitoneal dose required to kill 50% of a batch of 18 to 20 g of female Swiss-Webster mice (LD50 ) over 3 to 4 days.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567077#R105567077" 77] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567079#R105567079" 79] The original assay measured the LD50 in monkeys, which was found to be less narrow than the lethal dose effects in mice.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567007#R105567007" 7] The LD50 of BTX-A for humans, extrapolated from experiments in monkeys, has been estimated to be about 40 MU/kg.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567080#R105567080" 80] For a 70-kg man, the LD50 falls in the range of 2500 to 3000 MU. Species variability in their sensitivity to the toxin prevents an accurate calculation of the toxic dose for humans.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567033#R105567033" 33] Nevertheless, there is a wide safety margin in therapeutic use. For example, the average number of units required for the treatment of glabellar frown lines is about 30 MU. The greatest pitfall of the mouse bioassay is that it does not provide an accurate characterization of the potency of BTX in humans.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567081#R105567081" 81] Clinical potency is dependent upon the toxin's dose, volume, and targeted muscle. Immunoassays Because of the expenses for animals and testing facilities and issues of ethical consideration, in vitro immunoassays (immunodiffusion assay, hemagglutination assay, radioimmunoassay, enzyme-linked immunosorbent assay [ELISA]) have also been developed for rapid detection and quantitation of BTX. Immunodiffusion and hemagglutination assays, which use a single monoclonal antibody, are still not sensitive enough to replace the mouse bioassay.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567082#R105567082" 82] The requirements to radiolabel toxin and to have suitable radiologic facilities make radioimmunoassay inapplicable for the routine testing of samples. ELISA has the greatest potential as a replacement for the bioassay. The standard ELISA technique using polyclonal antibodies is highly specific, reasonably rapid, and can be applied to the testing of a large number of specimens.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567074#R105567074" 74] The sensitivity of ELISA can be further improved by a redox cycle amplification system [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567083#R105567083" 83] or an enzyme-linked coagulation amplification system.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567084#R105567084" 84] The major disadvantage of all ELISA-based assays is the inability to differentiate between active and inactive toxin.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567074#R105567074" 74] A specific assay based on the BTX endopeptidase activity is being investigated. Unlike ELISA-based assays, this new assay directly measures the biologic activity of the toxin.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567085#R105567085" 85] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567086#R105567086" 86] These in vitro assays may not totally replace the mouse bioassay because they do not give a measurement of other parameters such as cell binding and internalization, which contribute to overall toxicity and therapeutic potency. Therefore the mouse bioassay is currently the gold standard for toxin detection and standardization. Determinants of clinical response There is no clinical standardization on the use of BTX-A. Factors that affect clinical response are commercial preparations, anatomy, dose and response relationship, storage, and immunogenicity. Preparations There are two commercially available BTX-A preparations: Botox (Allergan, Inc, Irvine, Calif) and Dysport (Speywood Pharmaceuticals, Maidenhead, England). Only Botox is currently available in the United States. The Botox unit is 3 to 5 times as potent as the Dysport unit.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567014#R105567014" 14] The currently available batch of Botox, derived from toxins prepared by Allergan, Inc, in 1997, has replaced the old batch originally prepared by Schantz in 1979.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567087#R105567087" 87] Botox is a sterile, lyophilized (vacuum-dried) form of purified BTX type A, produced from a culture of the Hall strain of C botulinum grown in a medium containing N-Z amine and yeast extract. It is isolated from the culture solution by a series of acid precipitations to a crystalline complex consisting of the active high molecular weight toxin protein and an associated hemagglutinin protein. The crystalline complex is then redissolved in a solution containing saline and albumin for stability, and sterile-filtered before vacuum-drying. In the Schantz preparation, 1 MU of the crystalline protein complex weighed about 0.043 ng.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567078#R105567078" 78] The amount of chromatographically purified botulinum A toxin was approximately 0.006 ng.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567088#R105567088" 88] The new batch has only 20% of the protein content of the old batch. Each vial of Botox contains 100 MU of C botulinum toxin type A (with 10% variability), 0.5 mg of human albumin, and 0.9 mg of sodium chloride in a sterile, vacuum-dried form without a preservative. The average cost per vial is about $370. The vials are stored in the freezer before reconstitution for clinical use. The recommended diluent is nonpreserved normal saline. The use of preserved saline during reconstitution may alter the dose response.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567017#R105567017" 17] Excessive shaking and bubbling during reconstitution may inactivate the toxin. After reconstitution, the product should be stored in the refrigerator at 2░C to 8░C. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567089#R105567089" 89] Dysport (BTX type A) preparation is different in terms of MU, chemical properties, biologic activities, and weight.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567090#R105567090" 90] It is supplied in 500 MU vials, produced by column-based purification rather than by the precipitation technique used for Botox, and may be stored at room temperature. The recommended diluent is also unpreserved normal saline.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567091#R105567091" 91] It has been used successfully for blepharospasm,[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567090#R105567090" 90] torticollis,[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567092#R105567092" 92] hemifacial spasm, [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567093#R105567093" 93] and hyperfunctional facial lines. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567091#R105567091" 91] Anatomy Treatment should be tailored to the individual patient. The size and orientation of muscle fibers vary between different anatomic regions and sexes. Hyperfunctional lines are perpendicular to the sum vectors of muscle forces. All the superficial muscles of the face are part of the superficial musculoaponeurotic system.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567094#R105567094" 94] The muscles in the glabellar region are intertwined. The medial fibers of the frontalis muscles are the only medial brow elevators. The medial brow depressors consist of the corrugator supercilii, depressor supercilii, orbital portion of the orbicularis oculi, and procerus. The strong corrugator muscles contribute most to the frown lines and require higher doses of BTX for muscle paralysis.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567020#R105567020" 20] Paralysis of the medial brow depressor muscles reduces the vertical glabellar wrinkles and horizontal nasal root wrinkles with a concomitant medial eyebrow lift (Huang et al, manuscript accepted for publication). The frontalis muscle raises the medial and lateral brows and contributes to horizontal forehead line wrinkles. Because the frontalis spans the entire forehead, multiple point injections are required for effective paralysis.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567095#R105567095" 95] The medial fibers, being more fibrous than the lateral fibers, require larger doses. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567094#R105567094" 94] Injection of lower frontalis fibers too close to the lateral brow may lead to brow ptosis.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567021#R105567021" 21] The function of the orbicularis oculi muscle is to close the eye. Contraction of the lateral orbicularis oculi muscle gives rise to crow's feet. Because of the superficial position of the orbicularis oculi muscle, injection in this region is usually directed into the dermal or subcutaneous plane along the outer orbital rim peripheral to the lateral canthus. This technique also helps to minimize toxin diffusion and paralysis of neighboring muscles. For example, unintentionally paralyzing the levator labii superioris muscle would result in temporary eyelid ptosis. Innervation studies of the orbicularis oculi muscle have demonstrated a diffuse distribution of neuromuscular junctions.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567096#R105567096" 96] Therefore multiple injection points are more likely to provide a satisfying outcome than a total equivalent dose given into one injection point. Women's brows are frequently arched above the orbital rim. Men tend to have a more horizontal brow. In patients with horizontally oriented brows and deep frown lines, an injection 1 cm above the orbital rim in the midpupillary line may be needed to paralyze the tail of the corrugator and the temporal orbicularis. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567097#R105567097" 97] The depths of all dynamic lines may be augmented by photodamage. As a result, a complete correction of the lines may not be possible. However, with relaxation of the muscle and protection from photodamage, dermal remodeling may change with time. For a review of other anatomic Botox sites (lips, platysma, axilla, and palms and soles), the reader should refer to more in-depth articles.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567023#R105567023" 23] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567025#R105567025" 25] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567027#R105567027" 27] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567028#R105567028" 28] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567098#R105567098" 98] Dose and response relationships The effect of Botox is dependent on the location, concentration, and volume of solution that is injected. The art of using Botox comes from choosing the desired weakening of muscle contraction without causing unwanted muscle paralysis. Reported concentrations of solutions used for cosmetic indications range from 1 MU/0.1 to 10 MU/0.1 mL. Reported volumes injected in each location range from 0.025 to 1.0 mL per site. Typically volume and concentration are increased to correlate with the size of the treated muscle. Dosing in general is still rather arbitrary and based on the experience of the physician.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567099#R105567099" 99] In attempt to examine the relationship between dose, volume, and targeted muscles, Borodic et al[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567100#R105567100" 100] used rabbit longissimus dorsi and acetylcholinesterase staining as indices of denervation and showed that the size of the denervation field is determined by dose and volume. Specifically, a single injection of 10 MU per 0.1 mL causes a toxin spread of 4.5 cm. Using rat anterior tibialis muscle and periodic acid-Schiff staining to measure glycogen depletion and quantify paralysis, Shaari and Sanders[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567101#R105567101" 101] found that dose was a stronger predictor of area of paralysis than volume and the effect of BTX was greatest when injected closest to the motor end plate. Injecting into the crow's feet area with a total dose of 6 to 15 MU with 3 injection sites, Carruthers[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567102#R105567102" 102] noted the effect of the toxin spread to be at least 1 cm. Using the glabellar frown lines as a model to evaluate the dose-response, Hankins, Strimling, and Rogers[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567103#R105567103" 103] concluded that the threshold for a demonstrable response to BTX is between a total dose of 5 and 12.5 MU or 1 to 2.5 MU per injection site (a total of 5 injection sites), whereas an effective starting dose is between a total dose of 12.5 and 20 MU or 2.5 to 4 MU per injection site, with a duration of 2 to 5 months. There was no statistically significant difference in safety or efficacy for concentrations ranging from 50 to 200 MU/mL of BTX. In summary, to achieve maximal dose response and minimize side effects, the clinician should use the most effective dose at the smallest volume. At a particular injection site, small volume and high dose are superior to large volume and low dose. Small volume and high dose tend to localize the toxin and contain the biologic effect of muscle paralysis.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567095#R105567095" 95] Large volume and low dose weaken the muscle and may produce an overall smoothing effect[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567020#R105567020" 20] with a concomitant risk of toxin spread to adjacent muscles.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567022#R105567022" 22] Storage Storage after reconstitution may affect the biologic potency of Botox. The current Food and Drug Administration-approved product labeling recommends that it should be used within 4 hours of reconstitution with normal saline,[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567089#R105567089" 89] which makes Botox use somewhat cumbersome in clinical settings. Several studies have investigated the relationship between storage and potency. One study using the mouse bioassay showed no loss of activity 6 hours after reconstitution at room temperature. However, when left for 12 hours, a loss of up to 44% activity was observed. Refreezing the toxin after reconstitution was reported to cause approximately 70% loss of bioactivity after 1 to 2 weeks.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567104#R105567104" 104] In a human extensor digitorum brevis model, Sloop, Cole, and Escutin[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567105#R105567105" 105] showed no loss of potency in the reconstituted toxin after refrigeration or refreezing for 2 weeks. Using time stored diluted Botox, Lowe[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567091#R105567091" 91] observed a 50% decrease in the efficacy of Botox to reduce hyperfunctional facial lines after 1 week. Another study demonstrated that toxin at 10 MU/mL reconstituted 30 days before injection produced paralysis of facial muscle tone equivalent to that of freshly mixed toxin.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567020#R105567020" 20] All studies used unpreserved normal saline for reconstitution. Although there are no standardized guidelines for storage, most clinicians do not refrigerate the toxin for more than 1 week. Refrigeration for less than 24 hours is optimal, and refreezing is discouraged.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567106#R105567106" 106] Immunogenicity The toxin's immunologic properties can lead to the stimulation of antibody production, potentially rendering further treatments ineffective. The minimum dose and injection schedule required to induce antibody formation are unknown. Immunogenicity has been shown to be dependent on dose per injection session, cumulative dose, and frequency of administration. A study by Biglan et al[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567107#R105567107" 107] revealed absence of antibody formation in patients who received less than 50 MU per injection session. Gonnering[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567108#R105567108" 108] reported antibody response in patients with facial spasm syndromes receiving doses in the 150 to 300 MU range but not in those receiving doses of up to 52.5 MU per session over a period of 163 weeks. Testing antibody production in cervical and oromandibular patients, Jankovic and Schwartz[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567109#R105567109" 109] observed a statistically significant difference between patients with antibodies and a mean cumulative dose of 1709 MU and patients without antibodies and a mean cumulative dose of 1066 MU. Zuber et al[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567110#R105567110" 110] noted a 3% prevalence rate in focal dystonia patients receiving a greater than 50 ng (~1162 MU) cumulative dose. Study of torticollis patients treated with doses ranging from 150 to 300 MU demonstrated a 4.3% prevalence of neutralizing antibody, and also noted that BTX-resistant patients received more frequent injections and more booster injections 2 to 3 weeks after treatment.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567111#R105567111" 111] To date, antibody formation has not been reported in patients treated for blepharospasm or dermatologic uses.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567018#R105567018" 18] The most widely used test for antibody detection is the mouse neutralization assay (Northview Pacific Labs, Berkeley, Calif).[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567112#R105567112" 112] Antibodies from human serum and BTX-A are coadministered to mice. The binding of antibodies to toxin protects mice from the toxin's lethal effects, and this neutralization is quantified. The mouse test is time-consuming and expensive.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567033#R105567033" 33] More rapid immunoassays have also been used to detect the anti-BTX antibody. Their main drawbacks are lower specificity[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567113#R105567113" 113] and lack of correlation between detected antibodies and clinical resistance.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567084#R105567084" 84] A combination of the mouse neutralization assay and an immunoassay would probably provide the most sensitive and specific test. To minimize antibody resistance, one should use the smallest possible effective dose, use treatment intervals of at least 3 months, and avoid booster injections. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567111#R105567111" 111] Moreover, shorter intervals between injections may not produce as prolonged a functional effect as longer intervals. [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567114#R105567114" 114] Patients with BTX-A resistance may benefit from injections with BTX-B (Athena Neurosciences, San Francisco),[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567012#R105567012" 12] BTX-C (Speywood Pharmaceuticals),[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567115#R105567115" 115] or BTX-F (Speywood Pharmaceuticals).[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567116#R105567116" 116] Because these other serotypes have different potencies, their duration of effect would vary.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567040#R105567040" 40] BTX-B, BTX-C, and BTX-F are currently under clinical investigation; the only commercially available toxin for routine clinical use at present is BTX-A. Precautions Botox is a Pregnancy Category C drug, which means that animal reproduction studies have not been conducted and that the human teratogenic effects of Botox are unknown. To date, there have been no reports of teratogenicity.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567117#R105567117" 117] It has not been determined whether this drug is excreted in human milk. Use of BTX in pregnant and lactating women is contraindicated. Caution should be exercised when using BTX in children younger than 12 years old.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567089#R105567089" 89] Adverse effects After a decade of therapeutic application of the toxin, no anaphylaxis or deaths attributable to Botox have been reported.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567040#R105567040" 40] Uncommon adverse effects are ptosis, ectropion, diplopia, eyelid drooping, hematoma, and bruising. Postinjection ptosis results from toxin diffusion through the orbital septum paralyzing the levator palpebrae superioris muscle.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567117#R105567117" 117] [ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567118#R105567118" 118] Transient ptosis is extremely rare and is usually minimal (1-2 mm of ptosis) and short-lived (lasting approximately 2 weeks).[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567117#R105567117" 117] Techniques to help avoid this complication include using low injection volume, accurately placing the needle 1 cm above the central eyebrow, aiming the needle upward and horizontally, injecting slowly to limit toxin spread, and exercising frequent muscle contraction for 30 minutes after the injection.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567021#R105567021" 21] Ptosis can be treated with apraclonidine 5% (Lopidine, Alcon, Inc, Dallas, Tex) but this should be done with caution. This alpha-adrenergic glaucoma medication causes contraction of the Muller's muscle, which is situated beneath the levator muscle of the upper eyelid, and results in elevation of the lash margin.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567097#R105567097" 97] Ectropion, diplopia, and drooping of the lateral lower eyelid can be avoided by injecting at least 1 cm peripheral to the bony orbit.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567097#R105567097" 97] Local hematoma and bruising are prevented by immediate digital pressure on the injection site. Other minor side effects are pain and temporary headaches.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567119#R105567119" 119] Long-term effects of Botox may include local changes in muscle fiber size and electromyographic abnormalities. These changes do not appear to have any clinical significance.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567040#R105567040" 40] There are no remote clinical effects of Botox, though local injection can produce subclinical electromyographic changes in uninjected distant muscles. The mechanism of this action is unclear.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567120#R105567120" 120] BTX does not cross the blood-brain barrier and therefore has no central nervous system effects.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567121#R105567121" 121] Whether intact BTX reaches the central nervous system after intramuscular injection by retrograde transport is undetermined and unlikely to be clinically important.[ HYPERLINK "http://home.mdconsult.com/das/article/body/1/jorg=journal&source=&sp=11464560&sid=0/N/183823/1.html" \l "R105567040#R105567040" 40] Conclusion This article reviews the pharmacology of BTX and its past and present uses. Botox has been shown to be a safe and effective treatment. Standardization of administrative techniques with emphasis on maximizing efficacy and minimizing complications will be useful in achieving consistent beneficial results in the future. As the use of BTX evolves, the field of dermatology will build upon this foundation of knowledge and explore more creative applications. References 1. Shone CC. Clostridium botulinum neurotoxins, their structures and modes of action. In: Watson ED, editor. Natural toxicants in foods. Chichester: Ellis Harwood Ltd; 1986. p. 11-57. а 2. Coffield JA, Bakry N, Zhang RD, Carlson J, Gomella LG, Simpson LL. In vitro characterization of botulinum toxin types A, C, and D action on human tissues: combined electrophysiologic, pharmacologic, and molecular biologic approaches. J Pharmacol Exp Ther 1997;280:1489-98. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/1189396?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 3. Hatheway CL. Botulism: the present status of the disease. Curr Top Microbiol Immunol 1995;195:55-75. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/635230?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 4. Macdonald KL, Cohen ML, Blake PA. The changing epidemiology of adult botulinum in the United States. Am J Epidemiol 1986;124:794-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7689572?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 5. Shapiro RL, Hatheway C, Swerdlow DL. Botulism in the United States: a clinical and epidemiologic review. Ann Intern Med 1998;129:221-8. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10348364?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 6. Merson MH, Dowell VR Jr. Epidemiologic, clinical, and laboratory aspects of wound botulism. N Engl J Med 1973;289:1105-10. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/4238355?PAGE=1.html&source=MI" \t "_top" Citation 7. Scott AB. Botulinum toxin injection into extraocular muscles as an alternative to strabismus surgery. Ophthalmology 1980;87:1044-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/6134419?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 8. Scott AB. Botulinum toxin injection of eye muscles to correct strabismus. Trans Am Ophthalmol Soc 1981;79:734-70. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/6534504?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 9. Scott AB, Kennedy RA, Stubbs HA. Botulinum A toxin injection as a treatment for blepharospasm. Arch Ophthalmol 1985;103:347-50. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7374994?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 10. Dutton JJ, Buckley EG. Long-term results and complications of botulinum A toxin in the treatment of blepharospasm. Ophthalmology 1988;95:1529-34. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/8192024?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 11. Savino PJ, Sergott RC, Bosley TM, Schatz NJ. Hemifacial spasm treated with botulinum A toxin injection. Arch Ophthalmol 1985;103:1305-6. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7434234?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 12. Tsui JK, Hayward M, Mak EK, Schulzer M. Botulinum toxin type B in the treatment of cervical dystonia: a pilot study. Neurology 1995;45:2109-10. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/610350?PAGE=1.html&source=HS,MI" \t "_top" Citation 13. Gelb DJ, Lowenstein DH, Aminoff MJ. Controlled trial of botulinum toxin injections in the treatment of spasmodic torticollis. Neurology 1989;39:80-4. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/8479857?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 14. Poewe W, Schelosky L, Kleedorfer B, Heinen F, Wagner M, Deuschl G. Treatment of spasmodic torticollis with local injections of botulinum toxin. One-year follow-up in 37 patients. J Neurol 1992;239:21-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1934661?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 15. Pasricha PJ, Ravich WJ, Hendrix TR, Sostre S, Jones B, Kalloo AN. Intrasphincteric botulinum toxin for the treatment of achalasia. N Engl J Med 1995;332:774-8. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/294319?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 16. Borodic GE, Cheney M, McKenna M. Contralateral injections of botulinum A toxin for the treatment of hemifacial spasm to achieve increased facial symmetry. Plast Reconstr Surg 1992;90:972-7, discussion 978-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2134595?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 17. Carruthers JD, Carruthers JA. Treatment of glabellar frown lines with C botulinum-A exotoxin. J Dermatol Surg Oncol 1992;18:17-21. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1927866?PAGE=1.html&source=CL,MI&ANCHOR=abs" \t "_top" Abstract 18. Carruthers A, Carruthers JDA. The use of botulinum toxin to treat glabella frown lines and other facial wrinkles. Cosmet Dermatol 1994;7:11-5. а 19. Foster JA, Barnhorst D, Papay F, Oh PM, Wulc AE. The use of botulinum A toxin to ameliorate facial kinetic frown lines. Ophthalmology 1996;103:618-22. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/707688?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 20. Garcia A, Fulton JE Jr. Cosmetic denervation of the muscles of facial expression with botulinum toxin: a dose-response study. Dermatol Surg 1996;22:39-43. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/648142?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 21. Carruthers A, Carruthers J. Cosmetic uses of botulinum A exotoxin. Adv Dermatol 1997;12:325-47; discussion 348. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/1154044?PAGE=1.html&source=MI" \t "_top" Citation 22. Edelstein C, Shorr N, Jacobs J, Balch K, Goldberg R. Oculoplastic experience with the cosmetic use of botulinum A exotoxin. Dermatol Surg 1998;24:1208-12. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10508427?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 23. Foster JA, Wulc AE, Holck DE. Cosmetic indications for botulinum A toxin. Semin Ophthalmol 1998;13:142-8. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10505995?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 24. Kane MA. Nonsurgical treatment of platysmal bands with injection of botulinum toxin A. Plast Reconstr Surg 1999;103:656-63; discussion 664-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10612943?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 25. Matarasso A, Matarasso SL, Brandt FS, Bellman B. Botulinum A exotoxin for the management of platysma bands. Plast Reconstr Surg 1999;103:645-52; discussion 653-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10612942?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 26. Frankel AS, Kamer FM. Chemical browlift. Arch Otolaryngol Head Neck Surg 1998;124:321-3. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10126974?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 27. Holmes S, Mann C. Botulinum toxin in the treatment of palmar hyperhidrosis. J Am Acad Dermatol 1998;39:1040-1. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10531777?ja=135547&PAGE=1.html&source=MI" \t "_top" Full Text 28. Glogau RG. Botulinum A neurotoxin for axillary hyperhidrosis: no sweat Botox. Dermatol Surg 1998;24:817-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10396284?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 29. Booij LH. Neuromuscular transmission and its pharmacological blockade. Part 1: neuromuscular transmission and general aspects of its blockade. Pharm World Sci 1997;19:1-12. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/9373948?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 30. Martin AR. Principles of neuromuscular transmission. Hosp Pract 1992;27:147-58. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2214790?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 31. Shone CC, Tranter HS. Growth of clostridia and preparation of their neurotoxins. Curr Top Microbiol Immunol 1995;195:143-60. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/635223?PAGE=1.html&source=MI" \t "_top" Citation 32. Sharma SK, Singh BR. Hemagglutinin binding mediated protection of botulinum neurotoxin from proteolysis. J Nat Toxins 1998;7:239-53. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10513003?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 33. Tsui JK. Botulinum toxin as a therapeutic agent. Pharmacol Ther 1996;72:13-24. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/1163980?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 34. Niemann H. Molecular biology of clostridial neurotoxins. In: Alouf JH, Freer JH, editors. A sourcebook of bacterial protein toxins. London: Academic Press; 1991. p. 303-48. а 35. Simpson LL. Molecular pharmacology of botulinum toxin and tetanus toxin. Annu Rev Pharmacol Toxicol 1986;26:427-53. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7986268?PAGE=1.html&source=MI" \t "_top" Citation 36. Black JD, Dolly JO. Interaction of 125I-labeled botulinum neurotoxins with nerve terminals. I: ultrastructural autoradiographic localization and quantitation of distinct membrane acceptors for types A and B on motor nerves. J Cell Biol 1986;103:521-34. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7657549?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 37. Black JD, Dolly JO. Interaction of 125I-labeled botulinum neurotoxins with nerve terminals. II: autoradiographic evidence for its uptake into motor nerves by acceptor-mediated endocytosis. J Cell Biol 1986;103:535-44. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/8914999?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 38. Schiavo G, Benfenati F, Poulain B, Rossetto O, Polverino de Laureto P, DasGupta BR, et al. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature 1992;359:832-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2223033?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 39. Li L, Singh BR. Isolation of synaptotagmin as a receptor for types A and E botulinum neurotoxin and analysis of their comparative binding using a new microtiter plate assay. J Nat Toxins 1998;7:215-26. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10513001?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 40. Brin MF. Botulinum toxin: chemistry, pharmacology, toxicity, and immunology. Muscle Nerve Suppl 1997;6:S146-68. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10509910?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 41. Hughes R, Whaler BC. Influence of nerve-ending activity and of drugs on the rate of paralysis of rat diaphragm preparations by Cl. botulinum type A toxin. J Physiol 1962;160:221-3. а 42. Nathan P, Dimitrijevic MR, Sherwood AM. Reflex path length and clonus frequency [letter]. J Neurol Neurosurg Psychiatry 1985;48:725. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7428139?PAGE=1.html&source=MI" \t "_top" Citation 43. Montecucco C, Schiavo G, Tugnoli V, de Grandis D. Botulinum neurotoxins: mechanism of action and therapeutic applications. Mol Med Today 1996;2:418-24. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/1089303?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 44. Sollner T, Whiteheart SW, Brunner M, Erdjument-Bromage H, Geromanos S, Tempst P, et al. SNAP receptors implicated in vesicle targeting and fusion. Nature 1993;362:318-24. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2359856?PAGE=1.html&source=CL,MI&ANCHOR=abs" \t "_top" Abstract 45. Huttner WB. Cell biology. Snappy exocytoxins. Nature 1993;365:104-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2649188?PAGE=1.html&source=MI" \t "_top" Citation 46. Santos JI, Swensen P, Glasgow LA. Potentiation of Clostridium botulinum toxin aminoglycoside antibiotics: clinical and laboratory observations. Pediatrics 1981;68:50-4. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/6134726?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 47. Wang YC, Burr DH, Korthals GJ, Sugiyama H. Acute toxicity of aminoglycoside antibiotics as an aid in detecting botulism. Appl Environ Microbiol 1984;48:951-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/6858895?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 48. Yamada S, Kuno Y, Iwanaga H. Effects of aminoglycoside antibiotics on the neuromuscular junction: Part I. Int J Clin Pharmacol Ther Toxicol 1986;24:130-8. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7624247?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 49. Snyder SH. Drug and neurotransmitter receptors: new perspectives with clinical relevance. JAMA 1989;261:3126-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/8614669?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 50. Simpson LL. The interaction between aminoquinolines and presynaptically acting neurotoxins. J Pharmacol Exp Ther 1982;222:43-8. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7154408?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 51. Kadieva V, van Heerden PV, Roux A, Friedman L, Morrell DF. Neuromuscular blockade and ventilatory failure after cyclosporine. Can J Anaesth 1992;39:402-3. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1955043?PAGE=1.html&source=MI" \t "_top" Citation 52. Fassi A, Sangalli F, Colombi F, Perico N, Remuzzi G, Remuzzi A. Beneficial effects of calcium channel blockade on acute glomerular hemodynamic changes induced by cyclosporine. Am J Kidney Dis 1999;33:267-75. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10622869?ja=202825&PAGE=1.html&source=MI" \t "_top" Full Text 53. Morel E, Feuillet-Fieux MN, Vernet-der Garabedian B, Raimond F, D'Anglejan J, Bataille R, et al. Autoantibodies in D-penicillamine-induced myasthenia gravis: a comparison with idiopathic myasthenia and rheumatoid arthritis. Clin Immunol Immunopathol 1991;58:318-30. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1554437?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 54. Vincent A, Newsom-Davis J. Acetylcholine receptor antibody characteristics in myasthenia gravis. II: patients with penicillamine-induced myasthenia or idiopathic myasthenia of recent onset. Clin Exp Immunol 1982;49:266-72. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/6313391?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 55. Bucknall RC. Myasthenia associated with D-penicillamine therapy in rheumatoid arthritis. Proc R Soc Med 1977;70 suppl 3:114-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/5730883?PAGE=1.html&source=MI" \t "_top" Citation 56. Albers JW, Hodach RJ, Kimmel DW, Treacy WL. Penicillamine-associated myasthenia gravis. Neurology 1980;30:1246-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7124937?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 57. Tzartos SJ, Morel E, Efthimiadis A, Bustarret AF, D'Anglejan J, Drosos AA, et al. Fine antigenic specificities of antibodies in sera from patients with D-penicillamine-induced myasthenia gravis. Clin Exp Immunol 1988;74:80-6. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/8685281?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 58. Walts LF. Neuromuscular blocking drugs. Otolaryngol Clin North Am 1981;14:501-13. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/6521128?PAGE=1.html&source=MI" \t "_top" Citation 59. Vincent A, Lang B, Newsom-Davis J. Autoimmunity to the voltage-gated calcium channel underlies the Lambert-Eaton myasthenic syndrome, a paraneoplastic disorder. Trends Neurosci 1989;12:496-502. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/8700855?PAGE=1.html&source=CL,MI&ANCHOR=abs" \t "_top" Abstract 60. Vincent A, Willcox N, Hill M, Curnow J, MacLennan C, Beeson D. Determinant spreading and immune responses to acetylcholine receptors in myasthenia gravis. Immunol Rev 1998;164:157-68. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10519561?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 61. Hoedemaekers AC, van Breda Vriesman PJ, De Baets MH. Myasthenia gravis as a prototype autoimmune receptor disease. Immunol Res 1997;16:341-54. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10042618?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 62. Simpson LL. Kinetic studies on the interaction between botulinum toxin type A and the cholinergic neuromuscular junction. J Pharmacol Exp Ther 1980;212:16-21. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7116065?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 63. Simpson LL. The origin, structure, and pharmacological activity of botulinum toxin. Pharmacol Rev 1981;33:155-88. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7233991?PAGE=1.html&source=MI" \t "_top" Citation 64. Bigotte L, Olsson Y. Cytotoxic effects of Adriamycin on mouse hypoglossal neurons following retrograde axonal transport from the tongue. Acta Neuropathol (Berl) 1983;61:161-8. а 65. Yamamoto T, Iwasaki Y, Konno H. Retrograde axoplasmic transport of Adriamycin: an experimental form of motor neuron disease? Neurology 1984;34:1299-304. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7319651?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 66. Gomez-Ramirez AM, Villegas-Perez MP, Miralles de Imperial J, Salvador-Silva M, Vidal-Sanz M. Effects of intramuscular injection of botulinum toxin and doxorubicin on the survival of abducens motoneurons. Invest Ophthalmol Vis Sci 1999;40:414-24. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10592853?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 67. Duchen LW. Changes in motor innervation and cholinesterase localization induced by botulinum toxin in skeletal muscle of the mouse: differences between fast and slow muscles. J Neurol Neurosurg Psychiatry 1970;33:40-54. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/3732015?PAGE=1.html&source=MI" \t "_top" Citation 68. Borodic GE, Ferrante R. Effects of repeated botulinum toxin injections on orbicularis oculi muscle. J Clin Neuroophthalmol 1992;12:121-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2074299?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 69. Alderson K, Holds JB, Anderson RL. Botulinum-induced alteration of nerve-muscle interactions in the human orbicularis oculi following treatment for blepharospasm. Neurology 1991;41:1800-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1713232?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 70. Borodic GE, Ferrante RJ, Pearce LB. Pharmacology and histology of the therapeutic application of botulinum toxin. In: Jankovic J, Hallet M, editors. Therapy with botulinum toxin. New York: Marcel Dekker; 1994. p. 119-57. а 71. Harris CP, Alderson K, Nebeker J, Holds JB, Anderson RL. Histologic features of human orbicularis oculi treated with botulinum A toxin. Arch Ophthalmol 1991;109:393-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1556570?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 72. Holds JB, Alderson K, Fogg SG, Anderson RL. Motor nerve sprouting in human orbicularis muscle after botulinum A injection. Invest Ophthalmol Vis Sci 1990;31:964-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1233477?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 73. Hassan SM, Jennekens FG, Wieneke G, Veldman H. Elimination of superfluous neuromuscular junctions in rat calf muscles recovering from botulinum toxin-induced paralysis. Muscle Nerve 1994;17:623-31. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/85250?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 74. Wictome M, Shone CC. Botulinum neurotoxins: mode of action and detection. Soc Appl Bacteriol Symp Ser 1998;27:87S-97S. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10494591?PAGE=1.html&source=MI" \t "_top" Citation 75. Hatheway CL, Ferreira JL. Detection and identification of Clostridium botulinum neurotoxins. Adv Exp Med Biol 1996;391:481-98. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/1021198?PAGE=1.html&source=MI" \t "_top" Citation 76. Pearce LB, First ER, MacCallum RD, Gupta A. Pharmacologic characterization of botulinum toxin for basic science and medicine. Toxicon 1997;35:1373-412. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10018726?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 77. Kautter DA, Solomon HM. Collaborative study of a method for the detection of Clostridium botulinum and its toxins in foods. J Assoc Anal Chem 1976;60:541-5. а 78. Schantz EJ, Kautter DA. Standardized assay for Clostridium botulinum toxins. J Assoc Anal Chem 1978;61:96-9. а 79. Schantz EJ, Johnson EA. Dose standardisation of botulinum toxin [letter]. Lancet 1990;335:421. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1522201?PAGE=1.html&source=HS,MI" \t "_top" Citation 80. Scott AB, Suzuki D. Systemic toxicity of botulinum toxin by intramuscular injection in the monkey. Mov Disord 1988;3:333-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/9036809?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 81. First ER, Pearce LB, Borodic GE. Dose standardisation of botulinum toxin [letter]. Lancet 1994;343:1035. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/178301?PAGE=1.html&source=MI" \t "_top" Citation 82. Tranter HS, Modi N, Hambleton P. New quality control methods of detecting bacterial toxins in food. In: Watson D, editor. Natural toxicants in food: progress and prospects. Chichester: Ellis Horwood and Sons; 1987. p. 169-203. а 83. Shone C, Wilton-Smith P, Appleton N, Hambleton P, Modi N, Gatley S, et al. Monoclonal antibody-based immunoassay for type A Clostridium botulinum toxin is comparable to the mouse bioassay. Appl Environ Microbiol 1985;50:63-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7585720?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 84. Doellgast GJ, Triscott MX, Beard GA, Bottoms JD, Cheng T, Roh BH, et al. Sensitive enzyme-linked immunosorbent assay for detection of Clostridium botulinum neurotoxins A, B, and E using signal amplification via enzyme-linked coagulation assay. J Clin Microbiol 1993;31:2402-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2510395?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 85. Shone CC, Quinn CP, Wait R, Hallis B, Fooks SG, Hambleton P. Proteolytic cleavage of synthetic fragments of vesicle-associated membrane protein, isoform-2 by botulinum type B neurotoxin. Eur J Biochem 1993;217:965-71. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2530404?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 86. Cornille F, Goudreau N, Ficheux D, Niemann H, Roques BP. Solid-phase synthesis, conformational analysis and in vitro cleavage of synthetic human synaptobrevin II 1-93 by tetanus toxin L chain. Eur J Biochem 1994;222:173-81. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/88483?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 87. Schantz EJ, Scott AB. Use of crystalline type A botulinum toxin in medical research. In: Lewis GE, editor. Biomedical aspects of botulinum. New York: Academic Press; 1981. p. 143-50. а 88. Hatheway CL. Toxigenic clostridia. Clin Microbiol Rev 1990;3:66-98. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1300949?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 89. Botox package insert. Irvine, Calif: Allergan Inc; 1995. а 90. Van den Bergh P, Francart J, Mourin S, Kollmann P, Laterre EC. Five-year experience in the treatment of focal movement disorders with low-dose Dysport botulinum toxin. Muscle Nerve 1995;18:720-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/412954?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 91. Lowe NJ. Botulinum toxin type A for facial rejuvenation: United States and United Kingdom perspectives. Dermatol Surg 1998;24:1216-8. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10508428?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 92. Odergren T, Hjaltason H, Kaakkola S, Solders G, Hanko J, Fehling C, et al. A double blind, randomised, parallel group study to investigate the dose equivalence of Dysport and Botox in the treatment of cervical dystonia. J Neurol Neurosurg Psychiatry 1998;64:6-12. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10053557?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 93. Jitpimolmard S, Tiamkao S, Laopaiboon M. Long term results of botulinum toxin type A (Dysport) in the treatment of hemifacial spasm: a report of 175 cases. J Neurol Neurosurg Psychiatry 1998;64:751-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10327463?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 94. Wieder JM, Moy RL. Understanding botulinum toxin: surgical anatomy of the frown, forehead, and periocular region. Dermatol Surg 1998;24:1172-4. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10508421?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 95. Borodic GE, Pearce LB, Smith K, Joseph M. Botulinum A toxin for spasmodic torticollis: multiple vs single injection points per muscle. Head Neck 1992;14:33-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2014355?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 96. Borodic GE, Cozzolino D, Ferrante R, Wiegner AW, Young RR. Innervation zone of orbicularis oculi muscle and implications for botulinum A toxin therapy. Ophthal Plast Reconstr Surg 1991;7:54-60. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1896995?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 97. Carruthers A, Carruthers J. Clinical indications and injection technique for the cosmetic use of botulinum A exotoxin. Dermatol Surg 1998;24:1189-94. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10508425?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 98. Hoefflin SM. Anatomy of the platysma and lip depressor muscles. Dermatol Surg 1998;24:1225-31. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10508430?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 99. Foster JA, Wulc AE. Cosmetic use of botulinum toxin. Facial Plast Surg Clin North Am 1998;6:79-85. а 100. Borodic GE, Ferrante R, Pearce LB, Smith K. Histologic assessment of dose-related diffusion and muscle fiber response after therapeutic botulinum A toxin injections. Mov Disord 1994;9:31-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/888956?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 101. Shaari CM, Sanders I. Quantifying how location and dose of botulinum toxin injections affect muscle paralysis. Muscle Nerve 1993;16:964-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2460498?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 102. Carruthers A, Carruthers J. Cosmetic uses of botulinum A exotoxin. In: Klein AW, editor. Tissue augmentation in clinical practice. New York: Marcel Dekker; 1998. p. 206-36. а 103. Hankins CL, Strimling R, Rogers GS. Botulinum A toxin for glabellar wrinkles: dose and response. Dermatol Surg 1998;24:1181-3. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10508423?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 104. Gargland MG, Hoffman HT. Crystalline preparation of botulinum toxin type A (Botox): degradation in potency with storage. Otolaryngol Head Neck Surg 1993;108:135-40. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2346226?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 105. Sloop RR, Cole BA, Escutin RO. Reconstituted botulinum toxin type A does not lose potency in humans if it is refrozen or refrigerated for 2 weeks before use. Neurology 1997;48:249-53. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/1126988?ja=75292&PAGE=1.html&source=HS,MI" \t "_top" Full Text 106. Carruthers A, Carruthers J. History of the cosmetic use of botulinum A exotoxin. Dermatol Surg 1998;24:1168-70. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10508420?PAGE=1.html&source=HS,MI" \t "_top" Citation 107. Biglan AW, Gonnering R, Lockhart LB, Rabin B, Fuerste FH. Absence of antibody production in patients treated with botulinum A toxin. Am J Ophthalmol 1986;101:232-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/7603903?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 108. Gonnering RS. Negative antibody response to long-term treatment of facial spasm with botulinum toxin. Am J Ophthalmol 1988;105:313-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/8067188?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 109. Jankovic J, Schwartz K. Response and immunoresistance to botulinum toxin injections. Neurology 1995;45:1743-6. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/501613?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 110. Zuber M, Sebald M, Bathien N, de Recondo J, Rondot P. Botulinum antibodies in dystonic patients treated with type A botulinum toxin: frequency and significance. Neurology 1993;43:1715-8. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2515645?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 111. Greene P, Fahn S, Diamond B. Development of resistance to botulinum toxin type A in patients with torticollis. Mov Disord 1994;9:213-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/891522?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 112. Hatheway CH, Snyder JD, Seals JE, Edell TA, Lewis GE Jr. Antitoxin levels in botulism patients treated with trivalent equine botulism antitoxin to toxin types A, B, and E. J Infect Dis 1984;150:407-12. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/6832290?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 113. Hanna PA, Jankovic J. Mouse bioassay versus Western blot assay for botulinum toxin antibodies: correlation with clinical response. Neurology 1998;50:1624-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10309730?ja=116678&PAGE=1.html&source=HS,MI" \t "_top" Full Text 114. Inagi K, Ford CN, Rodriquez AA, Schultz E, Bless DM, Heisey DM. Efficacy of repeated botulinum toxin injections as a function of timing. Ann Otol Rhinol Laryngol 1997;106:1012-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/10019475?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 115. Eleopra R, Tugnoli V, Rossetto O, Montecucco C, De Grandis D. Botulinum neurotoxin serotype C: a novel effective botulinum toxin therapy in humans. Neurosci Lett 1997;224:91-4. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/9387884?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 116. Sheean GL, Lees AJ. Botulinum toxin F in the treatment of torticollis clinically resistant to botulinum toxin A. J Neurol Neurosurg Psychiatry 1995;59:601-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/609323?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 117. Carruthers A, Kiene K, Carruthers J. Botulinum A exotoxin use in clinical dermatology. J Am Acad Dermatol 1996;34:788-97. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/720203?ja=51003&PAGE=1.html&source=MI" \t "_top" Full Text 118. Guyuron B, Huddleston SW. Aesthetic indications for botulinum toxin injection. Plast Reconstr Surg 1994;93:913-8. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/38714?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 119. Foster JA, Wulc AE, Barnhorst D, Papay F. The use of botulinum A toxin to ameliorate facial dynamic lines. Int J Aesthet Restor Surg 1996;4:137-44. а 120. Lange DJ, Rubin M, Greene PE, Kang UJ, Moskowitz CB, Brin MF, et al. Distant effects of locally injected botulinum toxin: a double-blind study of single fiber EMG changes. Muscle Nerve 1991;14:672-5. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/1691207?PAGE=1.html&source=HS,MI&ANCHOR=abs" \t "_top" Abstract 121. Coffield JA, Considine RV, Simpson LL. The site and mechanism of action of botulinum neurotoxin. In: Janckovic J, Hallett M, editors. Therapy with botulinum toxin. New York: Marcel Dekker Inc; 1994. p. 3-13. а 122. Schiavo G, Rossetto O, Catsicas S, Polverino de Laureto P, DasGupta BR, Benfenati F, et al. Identification of the nerve terminal targets of botulinum neurotoxin serotypes A, D, and E. J Biol Chem 1993;268:23784-7. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2533475?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 123. Schiavo G, Rossetto O, Benfenati F, Poulain B, Montecucco C. Tetanus and botulinum neurotoxins are zinc proteases specific for components of the neuroexocytosis apparatus. Ann N Y Acad Sci 1994;710:65-75. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/415394?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 124. Williamson LC, Halpern JL, Montecucco C, Brown JE, Neale EA. Clostridial neurotoxins and substrate proteolysis in intact neurons: botulinum neurotoxin C acts on synaptosomal-associated protein of 25 kd. J Biol Chem 1996;271:7694-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/719953?PAGE=1.html&source=CL,MI&ANCHOR=abs" \t "_top" Abstract 125. Blasi J, Chapman ER, Yamasaki S, Binz T, Niemann H, Jahn R. Botulinum neurotoxin C1 blocks neurotransmitter release by means of cleaving HPC-1/syntaxin. Embo J 1993;12:4821-8. а 126. Schiavo G, Shone CC, Rossetto O, Alexander FC, Montecucco C. Botulinum neurotoxin serotype F is a zinc endopeptidase specific for VAMP/synaptobrevin. J Biol Chem 1993;268:11516-9. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/O/2407162?PAGE=1.html&source=MI&ANCHOR=abs" \t "_top" Abstract 127. Yamasaki S, Hu Y, Binz T, Kalkuhl A, Kurazono H, Tamura T, et al. Synaptobrevin/vesicle-associated membrane protein (VAMP) of Aplysia californica: structure and proteolysis by tetanus toxin and botulinal neurotoxins type D and F. Proc Natl Acad Sci U S A 1994;91:4688-92. а  HYPERLINK "http://home.mdconsult.com/das/journal/view/N/85641?PAGE=1.html&source=CL,MI&ANCHOR=abs" \t "_top" Abstract .КЛМ▄▀рст2 3 4 5 6 7 8 И Й К Л М Н Х Ц Ш ║ ╔ ╩ ╦ ╠ ▌ ▐ ▀ р ° ∙ · № ¤ ■ ! 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