Botulin

Eur Arch Otorhinolaryngol, 1996, 253(8), 475 - 80
Changes in glial fibrillary acidic protein immunoreactivity in the rat facial nucleus following various types of nerve lesions; Laskawi R et al.; We report about changes on astrocytes in the facial nucleus of the rat following various types of peripheral nerve lesions . Astrocyte-specific glial fibrillary acidic protein (GFAP) was labeled by immunohistochemistry and served as a marker for these changes . Increased GFAP immunoreactivity was found in the facial nucleus on the lesioned side within 2-3 days after axotomy . This change lasted longer (up to 1 year) when axon regeneration was prevented or delayed by placing a metal clip on the proximal nerve stump . Lesion of the trigeminal nerve prior to axotomy reduced the degree of GFAP immunoreactivity . No side differences were observed after botulinum toxin application.

J Neurol Sci, 1996 Jan, 135(1), 74 - 7
Electrophysiological studies in patients with blepharospasm before and after botulinum toxin A therapy; Behari M et al.; Botulinum toxin (BTX) is the treatment of choice for blepharospasm . To investigate if the locally injected BTX influences the central nervous system and in particular the brain stem, amplitude and latency of compound muscle action potential (CMAP) of orbicularis oculi, blink reflex and brain stem auditory evoked potential were studied in 12 blepharospasm patients before BTX therapy, 9 patients after BTX therapy and 9 age and sex matched healthy controls . The mean amplitude of orbicularis oculi in patients was significantly lower before BTX therapy (p < 0.05) and after BTX therapy (p < 0.02) . The amplitudes of R1, R2 and contralateral R2 (CR) was higher in patients than controls but did not reach statistical significance . Following BTX therapy there was a reduction in amplitude of R1, R2 and CR (p < 0.001) and prolongation of latency of R2 and CR (p < 0.05) . The ratio of blink reflex amplitude/CMAP amplitude of orbicularis oculi showed a significant decline after BTX therapy (p < 0.01) . This small study suggests the involvement of brain stem pathways following BTX therapy in patients with blepharospasm.

Neuroscience, 1996 Jan, 70(2), 567 - 76
Differences in the multiple step process of inhibition of neurotransmitter release induced by tetanus toxin and botulinum neurotoxins type A and B at Aplysia synapses; Poulain B et al.; In order to gain insights into the steps (binding, uptake, intracellular effect) which differ in the inhibitory actions of tetanus toxin and botulinum neurotoxins types A or B, their temperature dependencies were investigated at identified cholinergic and non-cholinergic synapses in Aplysia . Upon lowering the temperature from 22 degrees C to 10 degrees C, extracellularly applied botulinum neurotoxin type A and B appeared unable to inhibit transmitter release whilst tetanus toxin exhibited a residual activity . Binding of each toxin to the neuronal membrane appeared virtually unaltered following this temperature change . By contrast, the intracellular effects of botulinum neurotoxin type B and tetanus toxin were strongly attenuated by temperature reduction whereas the inhibitory action of botulinum neurotoxin type A was only moderately reduced . Importantly, this discrepancy relates to the known proteolytic cleavage of different synaptic proteins by these two toxin groups . Since both the binding and intracellular activity of botulinum neurotoxin type A are minimally affected at 10 degrees C, its inability to inhibit neurotransmission at this low temperature when applied extracellularly indicated attenuation of its uptake . Due to the strict temperature dependence of the intracellular action of tetanus toxin and botulinum neurotoxin type B, but not A, an examination of the effects of changes in temperature on the internalization step was facilitated by the use of heterologous mixtures of the toxins' heavy and light chains . At 10 degrees C, heavy chain from tetanus toxin but not from botulinum neurotoxin type B mediated uptake of botulinum neurotoxin type A light chain . Collectively, these results provide evidence that, at least in Aplysia, the uptake mechanism for botulinum neurotoxin types A and B differs from that of tetanus toxin.

Zh Mikrobiol Epidemiol Immunobiol, 1996 Jan-Feb, (1), 11 - 3
{A new approach to the prevention and treatment of lesions caused by bacterial toxins: prospects for using inhibitors of ADP ribosylation}; Krasil'nikov II et al.; The damaging action of a number of bacterial toxins is determined by their capacity for blocking the specific functions of regulatory proteins of eukaryotic cells by ADP-ribosylation . Experiments, made with the use of type B botulinic toxin and 3,N-butyrylaminobenzamide as an example, have demonstrated that specific ADP-ribosylation inhibitors are capable of making up a new group of highly active antagonists of microbial toxins.

Eye, 1996, 10 ( Pt 3), 385 - 91
Factors influencing success and dose-effect relation of botulinum A treatment; Abbasoglu OE et al.; Botulinum toxin type A (BTA) treatment is an alternative to strabismus surgery . In this retrospective study the data on 45 esotropic and 49 exotropic patients with concomitant strabismus who were treated with BTA were analysed for dose-effect relationship, the effect of repeat doses and amblyopia on success of botulinum treatment . The esotropic patients were treated with a total of 80 and exotropic patients with 91 injections . The deviations were corrected within 5 degrees of straight in 33% of esotropic and 18% of exotropic patients . In esotropic patients the effect was dose dependent . This relation was not shown in exotropic patients . The repeat doses of BTA corrected the deviation to the same extent as the primary ones for both esotropic and exotropic patients.

Dev Biol Stand, 1996, 86, 11 - 8
The three Rs of Russell & Burch and the testing of biological products; Balls M et al.; The principles of humane animal experimentation proposed by Russell & Burch (1959), namely, replacement, reduction, and refinement, are now commonly known as the Three Rs . These principles are clearly embodied in Article 7 of Directive 86/609/EEC . It is instructive, therefore, to consider the priority currently attached to compliance with these principles and to the Directive in the development and control of biological products . Specific comments are made on the need for the application of the Three Rs in relation to the testing of human diphtheria and tetanus vaccines, human pertussis vaccine, inactivated veterinary Gumboro vaccine, veterinary Newcastle disease vaccine, veterinary clostridial vaccines and botulinum toxin . We pose three questions: a . Are the minimum numbers of animals already being used in this area? b . Is any unnecessary pain, suffering, distress or lasting harm being inflicted on the animals used? and c . What could and should be done about any shortcomings in current practice? Finally, the role of ECVAM in the promotion of the Three Rs within the European Union will be reviewed.

Mov Disord, 1996 Jan, 11(1), 79 - 81
Perioperative use of botulinum toxin for movement disorder-induced cervical spine disease; Adler CH et al.; Patients with cervical dystonia or tics of the nuchal muscles can develop serious cervical spine disease . We report a series of four patients who received botulinum toxin injections to control their movement disorders prior to their required surgery . One patient with cervical tic-induced radiculomyelopathy required botulinum toxin injection postoperatively to facilitate stabilization of the cervical fusion . Two patients with torticollis-induced cervical radiculomyelopathy, and one patient with dystonia-induced C5 fracture, had botulinum toxin injected preoperatively to facilitate postoperative recovery . Botulinum toxin appears to be a useful adjunct in the treatment of cervical movement disorders prior to or following surgery for associated cervical spine disease.

Mov Disord, 1996 Jan, 11(1), 27 - 31
Unilateral injection of botulinum toxin in blepharospasm: single fiber electromyography and blink reflex study; Girlanda P et al.; We studied six patients affected with blepharospasm (BSP) . We injected botulinum toxin (BTX) around only one eye and saline solution around the other . Clinical rating of BSP was performed . Single fiber electromyography (SFEMG), compound motor action potential (cMAP) at the orbicularis oculi muscle by stimulation of the facial nerve, blink reflex, and blink reflex recovery curve were recorded . All clinical and electrophysiological investigations were carried out before, and 1, 2, and 4 weeks after treatment . Evidence of bilateral clinical benefit was provided . Following therapy, facial cMAP decreased bilaterally and SFEMG revealed statistically significant changes on both sides while the excitability curve of blink reflex remained unmodified . The results confirm that BTX affects merely the neuromuscular junctions . The clinical and neurophysiological effects are present on both sides also for unilateral injection probably because of toxin spreading.

Am J Physiol, 1996 Jan, 270(1 Pt 2), R238 - 45
Botulinum-induced muscle paralysis alters metabolic gene expression and fatigue recovery; Gorin F et al.; We evaluated the physiological, histochemical, and biochemical consequences of inhibiting contractile activity in rat skeletal muscles with botulinum toxin A (BTX) . Contractile activity was entirely eliminated 12-18 h after a single, focal, intramuscular injection of BTX into the rat tibialis anterior muscle (TA) . Neuromuscular transmission remained completely inhibited for 10-12 days, then slowly recovered . BTX-treated muscles exhibited a lower resistance to both high- and low-frequency fatigue at 7 and 14 days after injection, but contractile force recovered more rapidly in treated TA after fatigue . Treated TA showed a twofold increase in the activity of the triglyceride hydrolase enzyme lipoprotein lipase (LPL) and a comparable increase in the relative abundance of LPL steady-state mRNA . In contrast, there was a 28% reduction in protein levels of the muscle isozyme of glycogen phosphorylase (MGP) and a 70% decrease in relative MGP transcript levels . Similar changes in relative transcript levels of LPL and MGP were observed in the predominantly fast-twitch extensor digitorum longus after BTX injection, but relative LPL and MGP mRNA levels were not altered in predominantly slow-twitch soleus . Histochemical evidence indicated that fast-twitch glycolytic fibers had increased lipid content . These biochemical alterations were reversed 120 days after BTX treatment despite persistent atrophy.

Neurochem Int, 1996 Jan, 28(1), 1 - 9
A unifying hypothesis for acetylcholine release; Israel M et al.; Mediatophore is the only nerve terminal membrane protein known to translocate acetylcholine upon calcium action . It is localized at the active zone . In this review we attempted to describe its role in relation to the vesicular and membrane protein complexes that are formed at the active zone . The model pictures a possible set of sequential steps that lead to exocytosis . The smallest quantal events are attributed to mediatophore opening momentarily, while synaptic vesicles synchronize release by controlling the calcium microdomain . A clear distinction is made between sub-quantal ACh release preserved after Botulinum toxin action, and exocytosis of vesicular contents . A cybernetic model for release and exocytosis related to protein interactions is presented for future works.

Pflugers Arch, 1996, 431(6 Suppl 2), R297 - 8
Upregulation of calcitonin gene-related peptide at mouse motor nerve terminals poisoned with botulinum type-A toxin; Meunier FA et al.; Calcitonin gene-related peptide (CGRP)-like immunoreactivity of motor nerve terminals was investigated at different times after local in vivo injection of botulinum type-A toxin (BoNT/A) close to the mouse levator auris longus muscle . CGRP expression in most of control nerve terminals was undetectable, but markedly increased during muscle paralysis and synaptic remodelling and, declined once functional recovery occurred.

Pflugers Arch, 1996, 431(6 Suppl 2), R283 - 4
Synaptotagmin II immunoreactivity in normal and botulinum type-A treated mouse motor nerve terminals; Juzans P et al.; The distribution of synaptotagmin II, a synaptic vesicle protein, was examined by immunohistochemistry at normal mouse motor nerve terminals and after botulinum type-A treatment . An immunoreactivity to synaptotagmin II was detected in both control and botulinum type-A treated motor nerve terminals including newly formed sprouts . These data, together with other reports showing the absence of synaptotagmin I at the neuromuscular junction, suggest that synaptotagmin II is the isoform involved in transmitter release at motor nerve terminals.

Eur Neurol, 1996, 36(1), 29 - 35
Botulinum toxin A in non-dystonic tremors; Henderson JM et al.; The present pilot study evaluated the effect of botulinum toxin A on primarily non-dystonic tremors using accelerometry in a single-blind, placebo-controlled design . Resting, postural, intention, or head tremor were assessed before and approximately 1 month after intramuscular saline and botulinum toxin A (25-50 U) respectively . Half of the patients showed > or = 30% placebo effect . Tremor in 10 of 17 patients (60%) studied improved further after botulinum toxin A (range 30-95%), exceeding the placebo effect by > or = 30% . Nine patients demonstrated clinically significant focal weakness in the extensor muscles after botulinum toxin A which interfered with fine movements . Patients were subdivided into PD-like and ET-like tremor(s) . Both groups experienced large placebo effects for resting tremor, with little or no further improvement after botulinum toxin A . The improvement in postural tremor after botulinum toxin A, of 40% in the PD-like and 57% in the ET-like groups, however, was approximately twice that of placebo . In conclusion, botulinum toxin A exerts a modest tremorlytic effect, however the dose, and its distribution over the sites injected, need to be optimised to minimise focal weakness.

ORL J Otorhinolaryngol Relat Spec, 1996 Jan-Feb, 58(1), 13 - 22
Changes in the phosphorylation of neurofilament proteins in facial motoneurons following various types of nerve lesion; Laskawi R et al.; This report defines the conditions for changes in the phosphorylation state of neurofilaments (NF) after facial nerve lesions . In adult control rats, few phosphorylated neurofilament (pNF) epitopes were stained (using SMI 31 antibodies) in a small subpopulation of facial motoneurons . After various types of mechanical lesion (nerve transection with and without attaching a metal clip to the proximal nerve trunk, nerve crush, combined trigeminal and facial nerve lesions) of the right facial nerve, pNF immunoreactivity transiently increased in most cell bodies of facial motoneurons on the operated side . This pNF 'reaction' started within 2 days after the operation and persisted up to 2 weeks but remained longer in those animals in which axonal regeneration had been prevented or delayed by attaching a metal clip to the proximal nerve stump . After botulinum toxin application into facial muscles (i.e . inhibition of synaptic transmission at motoric endplates) there was no increase in the amount of pNF after 4 and 10 days, but it appeared in facial nuclei 4 weeks after injection on both, the treated and the untreated side, i.e . during functional restitution . Selectively transecting the 'vibrissal part' of the trigeminal nerve induced no obvious changes in the pNF immunoreactivity in facial motoneurons, but a combined trigeminal-facial lesion did . Labeling nonphosphorylated epitopes (using SMI 32 antibodies) showed a slight decrease in immunoreactivity in the neuropil of the facial nucleus 15 days after nerve transection and fixing a metal clip on the proximal nerve stump.

Neurologia, 1996 Jan, 11(1), 34 - 6
{Botulinum A toxins in the treatment of spasticity in cerebral palsy during childhood}; Garcia Ruiz PJ et al.; We evaluated the safety and efficacy of Botulinum- A toxin (BTX) in patients with equinus deformity associated with cerebral palsy (CP) . We prescribed BTX to six patients (all of whom had previously participated in a multicenter, randomized, double blind study of the same drug); treatment continued for at least 15 months . Four children showed striking improvement, being converted from toe-toe to consistent or occasional heel-toe gait . No side effects were observed . BTX appears to be safe and effective in patients with CP.

Pflugers Arch, 1996 Jan, 431(3), 325 - 34
On the possible origin of giant or slow-rising miniature end-plate potentials at the neuromuscular junction; Sellin LC et al.; Giant or slow-rising miniature end-plate potentials (GMEPPs) caused by vesicular release of acetylcholine (ACh) occur at any time in about 50% of mouse diaphragm neuro muscular junctions, but generally at frequencies less than 0.03 s-1 . Their frequency is, unlike that of miniature end-plate potentials (MEPPs), not affected by nerve terminal depolarization . Unlike MEPPs and stimulus-evoked end-plate potentials, GMEPPs have a prolonged time-to-peak and show an increase in time-to-peak with amplitude . By using these differences in amplitude and time course, GMEPPs can be separated from MEPPs . In contrast to MEPPs, GMEPPs are not blocked by botulinum neurotoxin type A . GMEPPs have a greater temperature sensitivity than MEPPs, disappearing at temperatures below 15 degrees C . Long-term paralysis by botulinum toxin and certain drugs which inhibit protein kinase C or affect actin filament polymerization (cytochalasins) enhance the frequency of GMEPPs . End-plate current recordings show that similar postsynaptic ACh receptors are activated by MEPPs and GMEPPs . It is suggested that GMEPPs are not caused by mechanisms involved in regulated neurotransmitter release but are generated by constitutive secretion.

Neurology, 1996 Jan, 46(1), 26 - 9
Botulinum toxin therapy, immunologic resistance, and problems with available materials; Borodic G et al.; Botulinum toxin is a valuable technology for the treatment of regional movement disease . High-dose applications ( > 100 LD50 units per injection cycle) have been associated with sensitization that renders further therapeutic injections ineffective . The true incidence of sensitization is probably underestimated by the mouse bioassay . Other immunotypes of botulinum toxin have been effective in producing some therapeutic benefit; however, duration of action (botulinum toxin type F) and lower potencies may make these less attractive alternatives than botulinum type A . Increased specific activity botulinum toxin may be a method to reduce antigen exposure and mitigate against immunoresistance associated with dystonia therapy . Limiting the dose to < or = 100 LD50 units per injection cycle may limit this complication in the interim.

Dermatol Surg, 1996 Jan, 22(1), 39 - 43
Cosmetic denervation of the muscles of facial expression with botulinum toxin . A dose-response study; Garcia A et al.; BACKGROUND . Botulinum toxin has been used for facial hemispasm, strabismus, and blepharospasm . Recently it has been advocated to treat the frown lines . We have extended this program to treatment of other muscles of facial expression . METHODS . Botulinum toxin is injected into the muscles of facial expression in two or three sessions to produce a temporary loss of muscle tone . A standard method of cooling, injection, and compression was developed to minimize pain and bruising . OBJECTIVE . To complete a dose-response study to document the optimum timing and amount of toxin needed for each muscle group . RESULTS . Two to five Botulinum toxin units per muscle was as adequate as higher doses . Toxin that was reconstituted 30 days earlier produced the same loss of muscle tone as freshly mixed toxin . With two or three injection sessions loss of muscle tone lasted for up to 1 year . CONCLUSION . Botulinum toxin is highly effective as an adjuvant therapy for facial rejuvenation . This minor surgical procedure can temporarily reduce the lines on the upper face and produce a pleasing effect . With proper dosing and dilution this rejuvenation program becomes cost effective.

Ann Otol Rhinol Laryngol, 1996 Jan, 105(1), 33 - 42
Long-term effects of botulinum toxin injections in spasmodic dysphonia; Davidson BJ et al.; The purpose was to examine whether physiological changes can be found in laryngeal muscles following repeated treatment with botulinum toxin injections in spasmodic dysphonia . Seven patients whose treatment consisted of multiple unilateral thyroarytenoid injections were examined more than 6 months following their most recent botulinum toxin injection fiberoptic laryngoscopy and electromyography . Comparisons were made between injected and contralateral noninjected muscles' motor unit characteristics, muscle activation patterns, and vocal fold movement characteristics . The results demonstrated that motor unit characteristics differed between injected and noninjected muscles and that these differences were greater in patients less than 12 months since last injection . Motor unit duration differences were reduced and motor unit amplitude and numbers of turns were increased in muscles sampled over 1 year after injection . These results suggest that while the physiologic effects of botulinum toxin are reversible, the reinnervation process continues past 12 months following injection.

Laryngoscope, 1996 Jan, 106(1 Pt 1), 86 - 92
The swallowing side effects of botulinum toxin type A injection in spasmodic dysphonia; Holzer SE et al.; Botulinum toxin type A (BOTOX) injection of the thyroarytenoid muscle is used to control speech symptoms in patients with adductor spasmodic dysphonia . Transient difficulty in swallowing liquids is a common treatment side effect . Laryngeal movement durations were measured during swallowing in 13 adductor spasmodic dysphonia patients undergoing treatment and in 6 normal control subjects in order to determine the following: 1 . whether, prior to the injection, laryngeal movement durations were longer in the spasmodic dysphonia patients than in the control subjects; 2 . whether movement durations increased following the injections; 3 . whether preinjection swallowing difficulties related to postinjection swallowing measurements and postinjection patient reports of swallowing problems . A piezoelectric movement transducer was shown to be accurate for noninvasive measurement of laryngeal movement duration in relation to muscle onset and offset for hyoid elevation and relaxation . Before botulinum toxin type A injection, no significant differences in swallowing duration were found between the patient and control groups . Four patients with swallowing complaints prior to injection had longer laryngeal movement durations than the other spasmodic dysphonia patients and the control subjects . Following injection, laryngeal movement durations increased in the patients with spasmodic dysphonia, and eight patients reported dysphagia for an average of 2 weeks . Relationships were found between the patients' initial reports of swallowing problems and increased laryngeal movement durations before and after botulinum toxin type A injection . Those patients initially reporting swallowing difficulties had severe dysphagia for 2 weeks after the injection . Patient reports of dysphagia prior to injection may indicate a greater likelihood of significant dysphagia following thyroarytenoid injection with botulinum toxin type A.

Neuroreport, 1995 Dec 29, 7(1), 73 - 6
SNAP-25 is the major immunoreactive component of the brain-specific D3 protein; Jorgensen OS; The rat synaptosomal membrane antigen D3, which has been studied for more than 20 years since its first demonstration as a brain-specific protein, was shown to bind monoclonal antibodies to SNAP-25, a presynaptic plasma membrane-associated protein . In addition to SNAP-25, anti-D3 antiserum also precipitated two polypeptides of 34-36 kDa and 14-16 kDa . This suggests that anti-D3 binds all three of the 20S synaptic vesicle fusion particle components, i.e . SNAP-25, syntaxin, and vesicle-associated membrane protein (VAMP)/synaptobrevin . However, the botulinum toxins C and D, which are endoproteases selective for syntaxin and VAMP/synptobrevin, respectively, were unable to remove the D3 immunoreactivity from synaptosomal membranes . Therefore, it is concluded that D3 immunoreactivity is directed only against SNAP-25.

Ophthalmology, 1995 Dec, 102(12), 2036 - 40
Long-term changes in duration of relief with botulinum toxin treatment of essential blepharospasm and hemifacial spasm; Ainsworth JR et al.; PURPOSE: To determine long-term changes in duration of relief with serial treatments of botulinum A toxin (BAT) used to treat benign essential blepharospasm and hemifacial spasm, in view of conflicting reports as to whether BAT has an increasing, decreasing, or an unchanging duration of effect over a long period of treatment . METHODS: Thirty-two patients with facial dyskinesia (20 with essential blepharospasm, 12 with hemifacial spasm) were followed between 5 and 9 years through a mean of 18 (range, 12-32) BAT treatments with prospective documentation of intervals of relief from symptoms . Repeated measures and linear regression analyses were used to determine trends in each group . RESULTS: Marked inter- and intrapatient variability was found in the length of effect of BAT . Statistical analysis showed no significant changes in mean duration of relief within each group (P = 0.65 for essential blepharospasm, 0.36 for hemifacial spasm) . There was a trend to slow decline in the interval of relief, especially in patients with an initial duration of effect greater than 150 days . No relation was found between duration of relief and age or sex of patient or grade and duration of disease before initial treatment . CONCLUSION: In the long term, the mean duration of relief from symptoms with BAT changes little over a period of serial treatments . Short-term fluctuations in the length of therapeutic effect did not indicate the development of a resistance to treatment.

Neurosci Lett, 1995 Dec 1, 201(1), 37 - 40
Short-term electrical stimulation enhances the effectiveness of Botulinum toxin in the treatment of lower limb spasticity in hemiparetic patients; Hesse S et al.; The study tested the spasmolytic effect of Botulinum toxin A in two groups of hemiparetic patients with lower limb spasticity: in the first group (n = 5) 2000 U Dysport were injected into the soleus, tibialis posterior and both heads of gastrocnemius muscles alone; the second (n = 5) received additional repetitive alternating electrical stimulation of M . tibialis anterior and plantar flexors for 30 min six times per day during the 3 days following the injection . Muscle tone, rated by the Ashworth spasticity score, and gait analysis including recording of vertical ground reaction forces, were assessed before and 4 weeks after injection . The combined treatment proved to be more effective with respect to the clinically assessed reduction of muscle tone, gait velocity, stride length, stance- and swing-symmetry (P < 0.05) . The result is discussed with reference to animal experiments demonstrating enhanced toxin uptake and accelerated onset of its paralytic effect by electrical stimulation.

Aliment Pharmacol Ther, 1995 Dec, 9(6), 599 - 604
Review article: the use of botulinum toxin in the alimentary tract; Albanese A et al.; New and future indications for the treatment of disorders of the alimentary tract using local injections of botulinum toxin are reviewed . Clinical experience shows that overactive smooth muscle sphincters may be weakened to treat disorders such as achalasia or chronic anal fissure . By contrast, injections placed into the sphincter of Oddi have proven less effective for postcholecystectomy pain syndrome . Experimental evidence suggests that food intake may be reduced by weakening the distal stomach with botulinum toxin . This approach may possibly lead to the treatment of obesity . There are some new possible indications for the use of botulinum toxin on the alimentary tract, and infantile hypertrophic pyloric stenosis seems to be the most promising new development.

Gastroenterologist, 1995 Dec, 3(4), 273 - 88
Achalasia; Achkar E; Achalasia is a primary esophageal motor disorder characterized by lack of esophageal peristalsis and poor lower esophageal sphincter (LES) relaxation . Clinically, achalasia manifests as progressive dysphagia to solids and liquids and mild weight loss . Predisposition to esophageal cancer is not prevalent, but certain tumors may mimic achalasia . The diagnosis of achalasia is relatively easy to make with a good history, radiography, and esophageal motility testing . The esophagogram reveals a typical bird-beak narrowing of the esophagogastric junction and esophageal dilation, the degree of which depends on the stage of the disease . Esophageal manometry reveals poor LES relaxation, aperistalsis, and often elevated intraesophageal pressure . Endoscopic examination is important to rule out malignancy as the cause of achalasia . The traditional treatment of achalasia is forceful dilation of the LES . Bougienage may be helpful in some cases . Pharmacological agents, such as nitroglycerin and calcium channel blockers, provide some relief by decreasing LES pressure . However, they are not a viable, long-term choice . Surgical myotomy offers slightly better results than pneumatic dilation, but it is accompanied by some increased gastroesophageal reflux . Laparoscopic and thoroscopic myotomy are in their infancy, and, if successful, they will make surgical treatment much more attractive . Intrasphincteric botulinum toxin injection is the newest form of therapy . Its safety and ease of administration are very encouraging, but long-term results are not available.

Rinsho Shinkeigaku, 1995 Dec, 35(12), 1390 - 3
{Pathophysiologies of dystonia and myoclonus--consideration from the standpoint of treatment}; Segawa M; Pathophysiologies of disorders with dystonia or myoclonus were studied by evaluating the effects of treatment . Naturally, the main lesion of the dystonia responding to levodopa is in the nigrostriatal dopamine neuron . The target of stereotaxic operations is ventrolateral palladium for postural dystonia and the nucleus ventralis oralis posterior (Vop) thalamus for action dystonia . Torsion dystonia with lesion in the striatum and/or the pallidum causes axial torsion, it may be postural through the descending pathway and action through Vop . Stereotaxic operations on these pathways have shown to be effective . Focal dystonia is a reflection of abnormal co-activation of cortical motor neurons, occurring in a particular voluntary movement . Botulinus toxin injected into the affected muscle should be effective . Of myoclonus with epilepsy, cortical reflex myoclonus or cortical induced reticular myoclonus responds to valproic acid . However, no antiepileptic drugs are effective on those with primary brainstem lesion . Reticular reflex myoclonus due to asphyxia responds to ventralis intermedius thalamotomy . Idiopathic myoclonus associated with dystonia is particular because it responds to ventrolateral thalamotomy . Myoclonus except for idiopathic myoclonus with dystonia is associated with atonic NREM suggesting dysfunction of the dorsal raphe serotonergic neuron or the brainstem nucleus reticularis gigantocellularis, the causative neuron for experimental uremic myoclonus . Treatment for these neurons is necessary.

J Otolaryngol, 1995 Dec, 24(6), 345 - 51
Comparison of botulinum toxin injection procedures in adductor spasmodic dysphonia; Adams SG et al.; This study compares the effects of unilateral and bilateral thyroarytenoid muscle injections of botulinum toxin in 50 patients with adductor spasmodic dysphonia . Patients were randomly assigned to two treatment groups of 25 patients each and a group of 15 normal control subjects was also included . Using a standard electromyographic guidance procedure, one patient group received unilateral thyroarytenoid muscle injections of 15 units of botulinum toxin, while the second patient group received bilateral thyroarytenoid muscle injections of 2.5 units of botulinum toxin on each side . Follow-up data were obtained at 2- and 6-week intervals . Acoustic and perceptual measures of vocal performance included maximum phonation time, fundamental frequency, standard deviation of fundamental frequency, jitter, shimmer, signal/noise ratio, voice break frequency, spasm severity rating, and vocal breathiness rating . Unilateral and bilateral group comparisons at 2-weeks postinjection revealed no significant difference on any of the measures examined . At 6-weeks postinjection, maximum phonation time was significantly lower in the bilateral group . All other measures failed to differentiate the two patient groups . These results suggest that standard unilateral and bilateral botulinum toxin injections provide equivalent degrees of improvement in the symptoms of spasmodic dysphonia . However, bilateral injections appear to be associated with a longer period of excessive phonatory airflow than do unilateral injections.

Fortschr Neurol Psychiatr, 1995 Dec, 63(12), 495 - 503
{Bladder dysfunctions in encephalomyelitis disseminata--drug and interventional therapeutic options}; Zwergel U et al.; Patients with disseminated encephalomyelitis have various urological presentations, ranging from pollakisuria to urge incontinence . After detailed evaluation (neuro-urological examination, urodynamic investigation) drug therapy and various interventional methods must be adapted to the individual manifestations . Patients with detrusor hyperreflexia are treated with oral anticholinergic agents (oxybutynin, trospium chloride, propiverine) . Patients with urinary retention are recommended to be managed with clean intermittent (self)-catheterisation . The various interventional therapeutic options (bladder denervation, electrostimulation, local treatment with botulinum toxin) and the surgical therapy (sacral deafferentation and anterior root stimulation, bladder neck closure and cystostomy, sphincterotomy or augmentation cystoplasty) must be reserved for special cases.

J Voice, 1995 Dec, 9(4), 460 - 5
Combined-modality treatment of adductor spasmodic dysphonia with botulinum toxin and voice therapy; Murry T et al.; A combined-modality treatment program consisting of botulinum toxin injection (Botox) and voice therapy was used to treat 17 subjects diagnosed with adductor spasmodic dysphonia (ADD SD) . Ten subjects with ADD SD served as the control and were given Botox only . Voice therapy after Botox injection was directed toward reducing the hyperfunctional vocal behaviors, primarily glottal overpressure at voice onset and anterior-posterior squeezing . The results indicated that subjects who underwent combined-modality treatment maintained significantly higher mean airflow rates for significantly longer periods . Moreover, there was a carryover effect in these patients when they received Botox only . Adductor spasmodic dysphonia is treated most effectively when intrinsic laryngeal muscle spasms are reduced or eliminated by Botox injection and extrinsic hyperfunctional vocal behaviors are treated with voice therapy.

J Cell Biol, 1995 Dec, 131(6 Pt 2), 1747 - 58
Calcium-regulated exocytosis is required for cell membrane resealing; Bi GQ et al.; Using confocal microscopy, we visualized exocytosis during membrane resealing in sea urchin eggs and embryos . Upon wounding by a laser beam, both eggs and embryos showed a rapid burst of localized Ca(2+)-regulated exocytosis . The rate of exocytosis was correlated quantitatively with successfully resealing . In embryos, whose activated surfaces must first dock vesicles before fusion, exocytosis and membrane resealing were inhibited by neurotoxins that selectively cleave the SNARE complex proteins, synaptobrevin, SNAP-25, and syntaxin . In eggs, whose cortical vesicles are already docked, vesicles could be reversibly undocked with externally applied stachyose . If cortical vesicles were undocked both exocytosis and plasma membrane resealing were completely inhibited . When cortical vesicles were transiently undocked, exposure to tetanus toxin and botulinum neurotoxin type C1 rendered them no longer competent for resealing, although botulinum neurotoxin type A was still ineffective . Cortical vesicles transiently undocked in the presence of tetanus toxin were subsequently fusion incompetent although to a large extent they retained their ability to redock when stachyose was diluted . We conclude that addition of internal membranes by exocytosis is required and that a SNARE-like complex plays differential roles in vesicle docking and fusion for the repair of disrupted plasma membrane.

Circ Res, 1995 Dec, 77(6), 1222 - 8
Cutaneous active vasodilation in humans is mediated by cholinergic nerve cotransmission; Kellogg DL Jr et al.; During heat stress, increases in blood flow in nonglabrous skin in humans are mediated through active vasodilation by an unknown neurotransmitter mechanism . To investigate this mechanism, a three-part study was performed to determine the following: (1) Is muscarinic receptor activation necessary for active cutaneous vasodilation? We iontophoretically applied atropine to a small area of forearm skin . At that site and an untreated control site, we measured the vasomotor (laser-Doppler blood flow {LDF}) and sudomotor (relative humidity) responses to whole-body heat stress . Blood pressure was monitored . Cutaneous vascular conductance (CVC) was calculated (LDF divided by mean arterial pressure) . Sweating was blocked at treated sites only . CVC rose at both sites (P < .05 at each site); thus, cutaneous active vasodilation is not effected through muscarinic receptors . (2) Are nonmuscarinic cholinergic receptors present on cutaneous arterioles? Acetylcholine (ACh) was iontophoretically applied to forearm skin at sites pretreated by atropine iontophoresis and at untreated sites . ACh increased CVC at untreated sites (P < .05) but not at atropinized sites . Thus, the only functional cholinergic receptors on cutaneous vessels are muscarinic . (3) Does cutaneous active vasodilation involve cholinergic nerve cotransmission? Botulinum toxin was injected intradermally in the forearm to block release of ACh and any coreleased neurotransmitters . Heat stress was performed as in part 1 of the study . At treated sites, CVC and relative humidity remained at baseline levels during heat stress (P > .05) . Active vasodilator and sudomotor responses to heat stress were abolished by botulinum toxin . We conclude that cholinergic nerve activation mediates cutaneous active vasodilation through release of an unknown cotransmitter, not through ACh.

Otolaryngol Head Neck Surg, 1995 Dec, 113(6), 668 - 70
Voice failure after tracheoesophageal puncture: management with botulinum toxin; Blitzer A et al.; Primary or secondary tracheoesophageal puncture with a speaking prosthesis has provided rehabilitation of speech in most patients after total laryngectomy . Persistent constrictor spasm is thought to be responsible for a small percentage of these patients' inability to speak with the prosthesis . Management of these patients has included bougienage and pharyngeal myotomy and/or pharyngeal neurectomy . Botulinum toxin injections of the cricopharyngeus muscle complex in six patients have been successfully used diagnostically and therapeutically for tracheoesophageal puncture failures . The assessment, technique, and results are discussed.

J Neurol Neurosurg Psychiatry, 1995 Dec, 59(6), 601 - 7
Botulinum toxin F in the treatment of torticollis clinically resistant to botulinum toxin A; Sheean GL et al.; Two reports have shown a Japanese preparation of botulinum toxin type F (BTX-F) to be an effective alternative for patients with torticollis who develop clinical resistance to botulinum toxin type A (BTX-A) . A group of patients with torticollis, comprising five secondary non-responders and one primary non-responder, were treated with a preparation of BTX-F produced in the UK (Speywood Pharmaceuticals) . A low dose of BTX-F (220 mouse units (MU) in total) was given into clinically affected neck muscles, followed six weeks later by an injection of a total of 520 MU . Antibodies to BTX-A (mouse protection assay) were present in all secondary non-responders but not in the primary non-responder . No patients developed atrophy after injection of Dysport BTX-A (40 MU) into the left extensor digitorum brevis muscle whereas pronounced atrophy occurred in all patients after injection of 40 MU of BTX-F into the right extensor digitorum brevis muscle . Three patients improved subjectively after treatment with 220 MU BTX-F and five (all secondary non-responders) after the subsequent dose of 520 MU (two considerably), with reduced Tsui scores, but group scores were only significantly changed after the higher dose . The primary non-responder remained unchanged after both doses of BTX-F . One patient reported mild dysphagia with 520 MU BTX-F . Mean duration of improvement with 520 MU BTX-F was five (range 4-6)weeks . Thus BTX-F provides benefit for BTX-A non-responders with few side effects but for a shorter period than BTX-A, possibly due to relative underdosing . As with BTX-A, biological sensitivity to BTX-F does not necessarily predict a clinical response.

Curr Opin Ophthalmol, 1995 Dec, 6(6), 27 - 33
Eye movement disorders; Liu GT et al.; The ophthalmic, neurologic, and neuro-ophthalmic literature over the past year have included a wide variety of interesting case reports, patient series, and reviews involving eye movement abnormalities . This review highlights some of the more important articles and how they contribute to our understanding, diagnosis, and treatment of these disorders . A few topics will receive particular emphasis . In patients with sixth nerve palsies, botulinum toxin injection of the ipsilateral medial rectus muscle has been advocated . Recent results suggest that this treatment has no beneficial effect in acute sixth nerve palsies, but it may have a role in chronic cases . Two groups of authors, each supplying retrobulbar botulinum toxin injection for patients with acquired nystagmus and debilitating oscillopsia, obtained mixed results . One group of patients was moderately satisfied, whereas in the other group, no patients elected to repeat the treatment because of side effects such as ptosis, diplopia, or discomfort from keratitis . Finally, skew deviation is becoming a more recognized cause of vertical double vision from a central or peripheral basis . Articles published recently showed that cyclodeviation may be seen in skew deviation, and that binocular cyclotorsion distinguishes this motility abnormality from a fourth nerve palsy, which exhibits monocular excyclotorsion.

Ger J Ophthalmol, 1995 Nov, 4(6), 363 - 7
Long-term treatment of blepharospasm with botulinum toxin type A; Nussgens Z et al.; The treatment of essential blepharospasm with botulinum toxin has been known for a decade and is becoming increasingly more popular . To our knowledge, only a few longterm studies in major patient populations have been published . Of a total of more than 1,600 patients, results of treatment were evaluated in 115 patients (31 men and 84 women) treated continuously for a minimum of 3 years and a maximum of 8 years (mean, 5.7 years) . Patients were divided into two groups . Group I represents the "good responders" and contains all patients who received only 4-10 injections over that time (n = 55) . Group II represents the "poor responders, " who received at least 20 injections over that period (n = 60) . Group I received a mean of 7.1 injections, whereas group II had a mean of 24.4 injections (total, 1,855) . In group I the beneficial effect lasted for an average of 14.6 weeks (range, 2-52 weeks), whereas group II had a mean beneficial effect for only 6.8 weeks (range, 0-18 weeks) . The time of efficacy remained statistically stable even in the case of frequent treatment (up to 36 injections in group II) . Systemic or severe long-lasting local side effects were never observed; the most frequent side effects were: group I-ptosis, 5.4%; tearing, 5.1%; double vision, 1.8%; and lid lag, 1.5%; group II-ptosis, 4.3%; tearing, 3.3% lid lag, 1.9%, and double vision, 1.6% . The treatment of essential blepharospasm with botulinum toxin is a very effective therapy with minimal and transient complications . It may be used for long-term treatment without showing a decrease in efficacy.

J Protein Chem, 1995 Nov, 14(8), 703 - 8
Proteolysis of synthetic peptides by type A botulinum neurotoxin; Schmidt JJ et al.; Type A botulinum neurotoxin catalyzed the hydrolysis of synthetic peptides based on the sequence of the 25-kD synaptosomal protein SNAP-25 . In each peptide, the toxin cleaved at a single glutaminyl-arginine bond corresponding to residues 197 and 198 of SNAP-25, confirming earlier reports on the enzymatic specificity of the toxin in synaptosomal preparations . Metal chelators inhibited catalysis, consistent with a metalloprotease activity . In contrast to tetanus toxin and other botulinum toxin serotypes, type A toxin hydrolyzed relatively short, 17- to 20-residue peptides . In the substrates, SNAP-25 residue 202 and one or more of residues 187-191 were required for efficient hydrolysis, but residues 167-186 and 203-206 were not . The highest rates of hydrolysis were found when the C-terminal residues of the peptides were amidated.

Rinsho Shinkeigaku, 1995 Nov, 35(11), 1210 - 3
{Muscle afferent block for the treatment of writer's cramp}; Sawamoto N et al.; A 29-year-old man suffered from dystonic writer's cramp for over three years . When he wrote, typed and did other tasks using right hand, dystonic involuntary movement triggered medial rotation of the arm, wrist extension and shoulder elevation . Medication, biofeedback, and botulinum injection were performed without much success . We tried to block the sensory input from muscles by using lidocaine and ethanol . We made injections of 0.5% lidocaine 50ml and 99% ethanol 5ml into muscles with abnormal activity at the frequency of twice a week for about six months . After the treatment, dystonic movement was remarkably improved and he was then able to write, type and perform other tasks with the right hand . Side effects included pain of the injection site, nausea and dizziness, which lasted for a few hours . This "muscle afferent block" did not cause muscle weakness . We speculate that muscle afferent plays a pivotal role in dystonia so that its blocking may be of clinical use.

Neuropharmacology, 1995 Nov, 34(11), 1379 - 85
Proteins functioning in synaptic transmission at the sensory to motor synapse of Aplysia; Skehel PA et al.; Over expression of Aplysia synaptotagmin in acutely dissected cholinergic neurons from the buccal ganglia, or in primary co-cultures of glutaminergic sensory neurons and motor neurons, causes a reduction synaptic transmission . Anti-sense oligonucleotide treatment of similar cultures produced an enhancement of synaptic transmission . The interaction between Aplysia VAMP/synaptobrevin and syntaxin is reconstructed using the yeast two hybrid system, and used to identify amino acid residues of VAMP/synaptobrevin that are required for this interaction . Point mutations around residue 50, close to the site of cleavage by botulinum toxins specifically disrupt the interaction with syntaxin . An additional VAMP/synaptobrevin binding protein, VAP33, is identified using the yeast two hybrid system . Intracellular injection of VAP33 specific antisera inhibits synaptic transmission in sensory-motor neuron co-cultures.

Dig Dis, 1995 Nov-Dec, 13(6), 337 - 55
Esophageal dilatation; Nostrant TT; Esophageal dilatation as a treatment option in patients with both benign and malignant esophageal strictures is described . Types of dilators available, techniques of passage, complications, redilation rates, and comparative studies between dilating systems are reviewed . The use of proton pump inhibitors to reduce the rates of redilation is discussed in view of the natural history of benign esophageal strictures . Comparisons among blind passage, fluoroscopic guidance and endoscopically directed dilatation are made and discussed in relation to cost effectiveness . Brief mention of new dilating systems including wall stents and wall-tension-sensing systems are made . Achalasia treatment with pneumatic dilatation is described in detail and compared to medication and surgical myotomy options . New treatments, including botulinum toxin injection into the lower esophageal sphincter, are briefly mentioned.

HNO, 1995 Nov, 43(11), 644 - 8
{Therapy of Frey syndrome with botulinum toxin A . Experiences with a new method of treatment}; Drobik C et al.; The effectiveness of botulinum toxin for the treatment of Frey's syndrome is demonstrated . Since December 1993, 14 patients with severe symptomatic gustatory sweating have been treated at the ENT Department, University of Gottingen . Botulinum toxin A (approximately 0.5 U/cm2) was injected intracutaneously into the affected skin area as determined by Minor's starch iodine test . Gustatory sweating in the treated skin area ceased completely within 2 days and did not reappear during the period of following (13 months maximum follow-up) . There were no side effects . Findings show that local botulinum toxin injections are a highly effective, safe and minimally invasive treatment for Frey's syndrome . Moreover, this could be a new therapeutic tool for other forms of hyperhidrosis.

J Biol Chem, 1995 Oct 20, 270(42), 24631 - 4
G alpha 12 and G alpha 13 stimulate Rho-dependent stress fiber formation and focal adhesion assembly; Buhl AM et al.; Rho, a member of the Ras superfamily of GTP-binding proteins, regulates actin polymerization resulting in the formation of stress fibers and the assembly of focal adhesions . In Swiss 3T3 cells, heterotrimeric G protein-coupled receptors for lysophosphatidic acid and gastrin releasing peptide stimulate Rho-dependent stress fiber and focal adhesion formation . The specific heterotrimeric G protein subunits mediating Rho-dependent stress fiber and focal adhesion formation have not been defined previously . We have expressed GTPase-deficient, constitutively activated G protein alpha subunits and mixtures of beta and gamma subunits in Swiss 3T3 cells . Measurement of actin polymerization and focal adhesion formation indicated that GTPase-deficient alpha 12 and alpha 13, but not the activated forms of alpha 12 or alpha q stimulated stress fiber and focal adhesion assembly . Combinations of beta and gamma subunits were unable to stimulate stress fiber or focal adhesion formation . G alpha 12- and alpha 13-mediated stress fiber and focal adhesion assembly was inhibited by botulinum C3 exoenzyme, which ADP-ribosylates and inactivates Rho, indicating that alpha 12 and alpha 13, but not other G protein alpha subunits or beta gamma complexes, regulate Rho-dependent responses . The results define the integration of G12 and G13 with the regulation of the actin cytoskeleton.

Rev Prat, 1995 Oct 15, 45(16), 2029 - 36
{Neuro-orthopedic and muscle tonus disorders in patients with spinal cord injuries}; Dizien O et al.; Heterotropic ossifications and contractures are frequent during evolution of spinal cord injury . Heterotopic ossifications occur in the first months following the acute state . Hip is the most common localisation . No preventive or curative treatment is available . At an early stage, clinical or instrumental (sonographic) diagnosis may prevent functional consequences . Muscular disbalance, incomplete physiotherapy, tonus disorders lead to contractures of limbs or trunk . They may have severe consequences on activities of dairy living . Surgical procedures may be necessary . Tonus disorders may resist to usual drugs . In this cases, intrathecal infusion of baclofen, injection of botulinus toxin, neurosurgery of orthopedic surgery may be useful.

Brain Res, 1995 Oct 2, 694(1-2), 191 - 9
Skeletal muscle contraction modulates carbonic anhydrase phenotype in adult mouse dorsal root ganglion neurons; Mayeux V et al.; Recently carbonic anhydrase (CA) activity was demonstrated in adult mammalian proprioceptive neurons of the lumbar dorsal root ganglion (DRG) . To assess if neuron-target interactions govern the neuronal CA phenotype, we examined how various experimental procedures which modify the interactions of these neurons with their central and peripheral targets, affect mouse L5 lumbar DRG CA activity . In normal mice and under central disconnection, carbonic anhydrase activity was detected in 30% of neurons . One day after sciatic nerve transaction the percentage of CA-positive neurons decreased to around 50% of that in controls, although both the total number of neurons per ganglion and glial CA content were unchanged . The pattern of CA activity then remained stable until at least 30 days post-operative . All experimental procedures used to block muscle contraction, including ventral rhizotomy, tenotomy, local application to the nerve of both tetrodotoxin and lidocaine or intramuscular injection of the botulinum toxin, produced a significant decrease in neuronal CA staining . Moreover, axonal transport block by vinblastine induced a decrease in CA-positive neurons . These results show that functional neuron-muscle interactions independent of DRG-spinal Cord influences contribute to the regulation of CA activity in lumbar DRG neurons . This modulation could be under the control of unidentified activity-dependent molecular mechanism involving stimuli through the skeletal muscle contraction, inducing in turn, the synthesis of a CA-regulating factor(s) retrogradely transported to the neuronal cell body and/or nuclei.

EMBO J, 1995 Oct 2, 14(19), 4705 - 13
Clostridial neurotoxins compromise the stability of a low energy SNARE complex mediating NSF activation of synaptic vesicle fusion; Pellegrini LL et al.; A 20S complex composed of the cytosolic fusion proteins NSF and SNAP and the synaptosomal SNAP receptors (SNAREs) synaptobrevin, syntaxin and SNAP-25 is essential for synaptic vesicle exocytosis . Formation of this complex is thought to be regulated by synaptotagmin, the putative calcium sensor of neurotransmitter release . Here we have examined how different inhibitors of neurotransmitter release, e.g . clostridial neurotoxins and a synaptotagmin peptide, affect the properties of the 20S complex . Cleavage of synaptobrevin and SNAP-25 by the neurotoxic clostridial proteases tetanus toxin and botulinum toxin A had no effect on assembly and disassembly of the 20S complex; however, the stability of its SDS-resistant SNARE core was compromised . This SDS-resistant low energy conformation of the SNAREs constitutes the physiological target of NSF, as indicated by its ATP-dependent disassembly in the presence of SNAP and NSF . Synaptotagmin peptides caused inhibition of in vitro binding of this protein to the SNAREs, a result that is inconsistent with synaptotagmin's proposed role as a regulator of SNAP binding . Our data can be reconciled by the idea that NSF and SNAP generate synaptotagmin-containing intermediates in synaptic vesicle fusion, which catalyse neurotransmitter release.

Toxicon, 1995 Oct, 33(10), 1383 - 6
Expression of a large, nontoxic fragment of botulinum neurotoxin serotype A and its use as an immunogen; LaPenotiere HF et al.; Using the polymerase chain reaction, a large fragment of botulinum toxin was placed in two expression systems, one designed to produce a fusion protein product and another designed to produce only the toxin fragment . Expression of the fragment in the latter system was inconsistent . Expression of the fusion protein was easily measurable by ELISA . Mice were vaccinated with crude fusion protein, then challenged with native toxin . Mice receiving two immunizations were partially protected from up to 1200 LD50, suggesting that this toxin fragment may be a good vaccine candidate to replace the currently used toxoid.

Neuropediatrics, 1995 Oct, 26(5), 249 - 52
Botulinum toxin in the treatment of cerebral palsy; Denislic M et al.; This paper reports the results of botulinum toxin A treatment in 13 children with cerebral palsy . All patients except one exhibited dynamic deformities in one hand or foot and changes in muscle tone of corticospinal and extrapyramidal origin . The primary purpose of the treatment was to improve the impaired skilled movements which resulted from dystonic limb posture and were the most disabling symptoms in the group studied . The study showed that the botulinum toxin treatment produced a significant improvement in functional disability in terms of amelioration of skilled hand movements and foot posture (p < 0.01) . The injections took effect a few days after dystonic muscle infiltration, and the mean duration of improvement was 3.1 months (2.0-4.0 months) . Side effects were negligible and transient; transitory muscle weakness was the most frequent . Botulinum toxin A provides a safe and effective adjuvant treatment of dystonic skilled movements and gait disorders in children with cerebral palsy.

Curr Opin Neurol, 1995 Oct, 8(5), 367 - 71
Toxic neuropathies; Mizisin AP et al.; Current publications describe neurotoxic effects of metals, pharmaceutical products, and environmental, biological and experimental substances . Agents such as lead remain the object of new epidemiological methods to identify victims . The use of methylcobalamin to ameliorate acrylamide toxicity is being explored . Antiarrhythmic and antilipidemic drugs continue to be associated with neuropathic effects but appear to be amenable to adjustments in dosage . To help control the neurotoxic effects of the chemotherapeutic drugs taxol and cisplatin analogs of adrenocorticotropic hormone and neurotrophic factors are being used . A new immunosuppressant, FK 506, has replaced cyclosporin to facilitate organ transplantation, but unwanted effects, including peripheral neuropathy, have been documented in some patients . In experimental studies, FK 506 has been reported to accelerate the rate of nerve regeneration . Botulinum toxin for the treatment of localized spastic disorders is a useful therapy, but training and supervision has been recommended by the American Academy of Neurology . Experimental toxins, such as imminodipropionitrile, continue to provide useful insights into physiologic mechanisms, including the axonal transport of cytoskeletal components.

J Neurochem, 1995 Oct, 65(4), 1712 - 20
Phosphorylation of VAMP/synaptobrevin in synaptic vesicles by endogenous protein kinases; Nielander HB et al.; VAMP/synaptobrevin (SYB), an integral membrane protein of small synaptic vesicles, is specifically cleaved by tetanus neurotoxin and botulinum neurotoxins B, D, F, and G is thought to play an important role in the docking and/or fusion of synaptic vesicles with the presynaptic membrane . Potential phosphorylation sites for various kinases are present in SYB sequence . We have studied whether SYB is a substrate for protein kinases that are present in nerve terminals and known to modulate neurotransmitter release . SYB can be phosphorylated within the same vesicle by endogenous Ca2+/calmodulin-dependent protein kinase II (CaMKII) associated with synaptic vesicles . This phosphorylation reaction occurs rapidly and involves serine and threonine residues in the cytoplasmic region of SYB . Similarly to CaMKII, a casein kinase II (CasKII) activity copurifying with synaptic vesicles is able to phosphorylate SYB selectively on serine residues of the cytoplasmic region . This phosphorylation reaction is markedly stimulated by sphingosine, a sphingolipid known to activate CasKII and to inhibit CaMKII and protein kinase C . The results show that SYB is a potential substrate for protein kinases involved in the regulation of neurotransmitter release and open the possibility that phosphorylation of SYB plays a role in modulating the molecular interactions between synaptic vesicles and the presynaptic membrane.

Int J Biochem Cell Biol, 1995 Oct, 27(10), 1065 - 78
ADP-ribosylation in adrenal glands: purification and characterization of mono-ADP-ribosyltransferases and ADP-ribosylhydrolase affecting cytoskeletal actin; Fujita H et al.; Mono-ADP-ribosylation in mammals is poorly understood . In this study, we purified four mono-ADP-ribosyltransferases and one ADP-ribosylhydrolase from rat adrenal medulla . The four purified mono-ADP-ribosyltransferases had molecular weights of 69,000 by gel filtration, pH optima of 8.0, and Kms for their action on NAD of about 20 microM . The four enzymes ADP-ribosylated to the alpha-subunit of heteromeric GTP-binding proteins . After tryptic digestion of alkylated actin mono-ADP-ribosylated by the purified mono-ADP-ribosyltransferases or botulinum C2 toxin, the two radioactive peptide patterns were identical . The purified ADP-ribosylhydrolase with mono-ADP-ribosylated actin as the substrate had a molecular weight of 61,000 on gel filtration, a pH optimum of 7.5, and a Km for mono-ADP-ribosylated actin of about 7 microM . The enzyme released ADP-ribose from ADP-ribosylated actin and the alpha-subunit of hetromeric GTP-binding proteins . Actin monomers mono-ADP-ribosylated by the four mono-ADP-ribosyltransferases did not form actin filaments after the addition of Mg2+ . After release of ADP-ribose by ADP-ribosylhydrolase, actin filaments formed on the addition of Mg2+, suggesting that the polymerization and depolymerization of cytoplasmic actin the adrenal chromaffin cells may be regulated by mono-ADP-ribosylation.

Tidsskr Nor Laegeforen, 1995 Sep 20, 115(22), 2777 - 9
{Hemifacial spasms . Causes, clinical findings and treatment}; Ringstad O et al.; Hemifacial spasm is a condition characterized by involuntary, episodic, synchronous contractions of muscles innervated by the facial nerve in one half of the face . The treatment can be medical or neurosurgical, but the most effective is injections of botulinum toxin . 13 patients who received this treatment for hemifacial spasm are described . The treatment of different forms of focal dystonias has been described before in this journal . The aetiology of hemifacial spasm is different from that of focal dystonia . It is probably due to compression of the facial nerve, causing ectopic generation of impulses . Treatment with botulinum toxin is shown to be effective also for this condition, but the dosage and the duration of effect differ from those reported for focal dystonias . This may be due to differences in the pathophysiological mechanisms of the two conditions.

Curr Opin Ophthalmol, 1995 Oct, 6(5), 86 - 99
Blepharospasm and related facial movement disorders; Patel BC et al.; The variable clinical features and the relatively good response of blepharospasm to botulinum-toxin type A are now well established . The etiology and pathophysiology of blepharospasm and related facial movement disorders are still poorly understood . Genetic and histopathologic studies over the last year have contributed to our understanding of this disease . The most significant progress has been made in the electromyographic studies of the the levator palpebrae and orbicularis oculi muscles . Subclassification based on the electromyographic abnormalities of these two muscles have begun to improve our understanding of the variable responses to botulinum-toxin type A . Further electromyographic studies may help identify the best sites of injection for optimal response and differentiate patients requiring limited or complete myectomy . The development of the limited myectomy has provided excellent functional and cosmetic results with quick recovery times in selected patients.

Curr Opin Ophthalmol, 1995 Oct, 6(5), 3 - 8
Visual function deficits in children; Carruthers JD; In the past year, new work on sensory testing in preverbal infants has expanded our knowledge in three areas: 1) visual acuity testing in infants in varying levels of illumination; 2) visual evoked cortical potential-evidence for binocular function in infantile esotropia; and 3) a new time-efficient method to evaluate color vision in newborns and infants . A newly reported sequel to loss of binocularity is the development of A and V pattern strabismus . In another study, loss of fusion, as with acute concomitant esotropia, was shown to be a sign of serious intracranial disease . Regarding motor visual function deficits, new treatments of infantile esotropia syndrome and nystagmus with botulinum toxin type A are presented . Regarding social issues, visual function deficit of abused children and infants born to drug-abusing mothers are presented as a timely reminder that child abuse is still an enormous social problem, and that the visual system is far from exempt from damage.

Mov Disord, 1995 Sep, 10(5), 574 - 9
Botulinum toxin: influence on respiratory heart rate variation; Claus D et al.; Remote adverse effects of local intramuscular botulinum toxin were investigated in a prospective follow-up study . Twenty-six patients with spasmodic torticollis were examined (18 women, eight men, 45 +/- 13 years) . Respiratory heart rate variation (HRV) was investigated by a computerized method . Different parameters were recorded (beats per minute, coefficient of variation, root mean square successive difference (RMSSD), spectral analysis, difference and quotient between maximum and minimum RR intervals, mean circular resultant) . After one intramuscular injection of 12.5 ng botulinum toxin (Porton Products Ltd., England), no significant influence on HRV was seen . After the second injection, a significant attenuation was seen of four parameters (coefficient of variation, Rmax - Rmin, Rmax divided by Rmin, mean circular resultant) that lasted up to several months . No clinically manifest remote side effects and no cardiac arrhythmia were seen for several months of botulinum toxin treatment . Our investigation proves an effect of local intramuscular botulinum toxin on autonomic cardiac innervation.

Arq Neuropsiquiatr, 1995 Sep, 53(3-A), 403 - 10
{Botulinum toxin in blepharospasm, in hemifacial spasm, and in cervical dystonia: results in 33 patients}; Novis SA et al.; The effects of botulinum toxin type A were studied in 33 patients with dystonia (12 blepharospasms, 10 hemifacial spasms and 11 spasmodic torticollis) . A rate scale was used to evaluate the severity of the dystonic movements, before and two weeks after each injection . Among blepharospasm patients, eight were female and four were male; the mean age was 57.7 years; the mean time of the disease duration was four years . Three had familial history for similar disease; nine were essential and three had used neuroleptic drugs (tardive dystonia) . The mean dose used was 51.3 U, with a mean time of beneficial effects of 2.8 months . For 22 injections and reinjections, 14 (63.7%) showed an excellent result, five (22.7%) good and three (13.6%) null . In the hemifacial spasm group, eight were female and two male; the mean age was 52.6 years; the mean time of the disease duration was 7.4 years; eight were essential and two post-paralytic . The mean dose used was 32 U . From the total of 15 injections and reinjections, all of them (100%) had an excellent result, with a mean time of beneficial effect of 3.4 months . Among the cervical dystonic patients, eight were male and three female; the mean age was 44.2 years; the mean time of the disease duration was 12.2 years; six had essential dystonia, three had used neuroleptic drugs and two had familial history for similar disease . The mean dose used was 238.6 U, with the mean duration of effect of 3.5 months . From the total of 20 injections and reinjections, 18 (90%) had good result, one (5%) mild and one (5%) null.(ABSTRACT TRUNCATED AT 250 WORDS)

Neurology, 1995 Sep, 45(9), 1743 - 6
Response and immunoresistance to botulinum toxin injections; Jankovic J et al.; Botulinum toxin antibodies (ABS) may be a reason why occasionally patients do not have a response to injections with botulinum toxin type A (BTX) . We tested 86 patients with cervical or oromandibular dystonia for the presence of BTX ABS; 20 were positive and 66 were negative . All patients who tested positive had no response to BTX injections on at least two consecutive treatment sessions . When compared with 22 randomly selected patients with negative BTX ABS results, the patients with positive BTX ABS tests had an earlier age at onset (mean age: 31.8 +/- 16.7 years versus 43.4 +/- 10.5; p < 0.05), higher mean dose per visit (249.2 +/- 32.5 U versus 180.8 +/- 68.7, p < 0.0005), and higher total cumulative dose (mean dose: 1,709 +/- 638 U versus 1,066 +/- 938; p < 0.01) . Four out of five patients with positive ABS tests later had a response to botulinum toxin type F injections . Of 26 patients with negative BTX ABS results who were tested because of poor response on at least one visit, 21 had good response after subsequent injection and five had no effect . Except for young age at onset and higher dosages, there were no other factors that could reliably predict which patients would become immunoresistant to BTX type A injections . Treatment with alternate serotypes may offer clinical benefit to this group of patients . Absence of detectable BTX ABS may occur in patients with poor response to BTX injections because of inadequate dosage, injections of inappropriate muscles, or poor sensitivity of the BTX ABS bioassay.

J Neurol Neurosurg Psychiatry, 1995 Sep, 59(3), 309 - 11
Pretarsal application of botulinum toxin for treatment of blepharospasm; Aramideh M et al.; The response to botulinum toxin type A was compared after two injection techniques in 45 patients with blepharospasm . Initially, patients were treated according to a triple injection technique; two injections into the upper eyelid and one injection into the lower eyelid . Subsequently, without altering the dose, the same patient group received two further injections into the pretarsal portion of the orbicularis oculi muscle of the upper lid . Triple injections were given in 227 treatments, of which 81% were successful . Mean duration of benefit was 8.5 weeks . Additional pretarsal injections were given in 183 treatment sessions . The number of successful treatments significantly increased, to 95% (P < 0.001), and the mean duration of benefit increased to 12.5 weeks (P < 0.001) . Ptosis occurred significantly less often after pretarsal injections (P < 0.01) . Patients with combined blepharospasm and involuntary levator palpebrae inhibition responded better to the pretarsal injection technique.

Ann Otol Rhinol Laryngol Suppl, 1995 Sep, 166, 426 - 8
Use of botulinum toxin to prevent facial nerve stimulation following cochlear implantation; Langman AW et al.; Facial nerve stimulation can develop because of the electric fields generated from an activated cochlear implant . This problem can usually be ameliorated by reducing or eliminating the current through the electrodes causing the undesirable stimulation . Occasionally there may be numerous electrodes that have to be deactivated in order to eliminate the facial nerve stimulation, and this may result in a marked reduction of auditory benefit to the cochlear implant user . Botulinum toxin causes transient neuromuscular blockade and has been shown to be effective in reducing or eliminating facial nerve hyperactivity due to a variety of causes . This report details the use of botulinum toxin in a patient with a cochlear implant who developed excessive facial nerve stimulation that necessitated the deactivation of 15 of the implant's 22 electrodes . After treatment with botulinum toxin, twice as many electrodes could be comfortably activated without causing facial nerve stimulation . The dynamic range was increased in 4 of the remaining 6 electrodes that were activated prior to treatment . The patient reported a marked improvement in sound quality and understanding of speech without lipreading as compared to her pretreatment performance . Treatment with botulinum toxin appears to be useful for patients who have excessive facial nerve stimulation after cochlear implantation.

No To Shinkei, 1995 Sep, 47(9), 857 - 62
{Dose-response relationship in the treatment of cervical dystonia with botulinum toxin type A (AGN 191622)--a phase II study}; Mezaki T et al.; Injection of botulinum toxin type A has been the treatment of choice for spasmodic torticollis for several years . Although previous reports demonstrate its effectiveness and safety, the treatment strategy has been empirical . The present study, using the freeze-dried crystalline botulinum toxin type A (AGN 191622; Allergan Inc., Irvine, CA), aimed to compare the efficacy among three treatment groups divided into low, medium and high dosage levels . Fifty-one patients who entered the study were grouped into low-dose (60 units/session), medium-dose (120 units/session) and high-dose (240 units/session) groups . Two patients (one in low-dose group and the other in high-dose group) were excluded from the assessment of efficacy because they dropped out in the early phase of the study . One experienced worsening of an existing psychosis and the other developed an acute respiratory infection . Injection sites were decided individually by palpation . If the clinical response was not satisfactory four weeks after an injection, the patient was re-injected with the same dose of toxin . The follow-up period was 14 weeks from the initial injection . The results showed that the high-dose group improved more than the other groups in the parameters of severity of symptoms and subjective benefit (p = 0.000) . Also, fewer injections were required in the high-dose group to achieve substantial clinical benefit . Although the mean reduction in Tsui's score was not statistically significant among the groups, the "marked improvement" was seen more frequently in the high-dose group (p = 0.033) . Unfavorable adverse effects including excessive weakness and dysphasia were always mild and transient.(ABSTRACT TRUNCATED AT 250 WORDS)

Nippon Ganka Gakkai Zasshi, 1995 Sep, 99(9), 1036 - 44
{Surgical and botulinum toxin treatment in two cases of abnormal retinal correspondence-exotropia with congenital homonymous hemianopsia}; Iwashige H et al.; We report 2 unusual cases of congenital occipital hemianopsia associated with abnormal retinal correspondence (ARC)-exotropia . Two Japanese males, 25 and 28 years of age, visited our hospital for surgical correction of manifest exotropia with dissociated vertical deviation and overaction of the superior oblique muscle . Visual field examination demonstrated homonymous hemianopsia with approximately 5 to 10 degrees of macular sparing . Visually evoked potential examination showed small amplitude similar to dissociated vertical deviation patients in all half-field stimulation . Electrooculogram examination demonstrated defective pursuit to the ipsilateral side . Some investigators have speculated that progressive exotropia compensates for homonymous hemianopsia and is a rare contraindication for strabismus surgery . To confirm the deterioration of motor and visual functions before surgery in these patients, we tried injections of botulinum A type toxin into the lateral rectus muscle ipsilateral to the hemianopsia . Contrary to the hypothesis, our patients had no change in their binocular visual fields and visual function . Finally, we performed recession of the lateral rectus muscle, so that both patients were satisfied with their ocular alignment, with no marked change of visual behavior.

Neurosci Lett, 1995 Aug 18, 196(1-2), 37 - 40
Botulinum toxin type A inhibits Ca(2+)-dependent transport of acetylcholine in reconstituted giant liposomes made from presynaptic membranes from cholinergic nerve terminals; Lopez-Alonso E et al.; Giant liposomes were made from a mixture of asolectin phospholipid vesicles and presynaptic plasma membranes isolated from Torpedo cholinergic nerve endings . Acetylcholine filled giant liposomes were able to release neurotransmitter upon stimulation by the Ca2+ ionophore A23187 and Ca2+ . Botulinum neurotoxin type A inhibited this Ca(2+)-dependent acetylcholine transport . Additionally, Botulinum toxin type A decreased membrane fluidity of liposomes . These results suggest that Botulinum toxin can interact directly with components of the presynaptic plasma membrane and inhibit acetylcholine translocation . Furthermore, since the reconstituted liposomes do not have synaptic vesicle components, the observed effects may account for the action of Botulinum toxin on the non-quantal release of acetylcholine from motor nerve terminals.

J Biol Chem, 1995 Aug 4, 270(31), 18216 - 8
The effect of botulinum neurotoxins on the release of insulin from the insulinoma cell lines HIT-15 and RINm5F; Boyd RS et al.; Western blotting of the insulin-secreting beta-cell lines HIT-15 and RINm5F with anti-SNAP-25 (synaptosomal associated protein of 25 kDa), anti-synaptobrevin, and anti-syntaxin 1 antibodies revealed the presence of proteins with the same electrophoretic mobility as found in neural tissue . Permeabilization of both of these insulinoma cell lines to botulinum neurotoxin A by electroporation resulted, after 3 days of culture, in the loss of approximately 90% of SNAP-25 immunoreactivity . A similar permeabilization of these cells with botulinum neurotoxin B resulted in the cleavage of approximately 90% of the synaptobrevin-like immunoreactivities . Botulinum neurotoxin F also cleaved approximately 90% of the synaptobrevin-like immunoreactivity in RINm5F cells . The permeabilization of both insulinoma cells to neurotoxin A resulted in a > 90% inhibition of potassium-stimulated, calcium-dependent insulin release . By contrast, permeabilization of the insulinoma cell lines to neurotoxin B resulted in only a approximately 60% inhibition of potassium-stimulated insulin release in HIT-15 cells, and neither neurotoxin B nor F caused inhibition in RINm5F cells . Thus HIT-15 and RINm5F cells contain the components of the putative exocytotic docking complex described in cells derived from the neural crest . In HIT-15 cells both SNAP-25 and synaptobrevin appear to be involved in calcium-dependent insulin secretion, whereas in RINm5F cells SNAP-25 but not synaptobrevin is involved.

J Otolaryngol, 1995 Aug, 24(4), 209 - 16
Effect of neuromuscular activity on the response to botulinum toxin injections in spasmodic dysphonia; Wong DL et al.; Spasmodic dysphonia (SD), a neurologic disorder characterized by involuntary vocal spasms during speech, has been effectively treated by injections of botulinum toxin (BT) into the laryngeal muscles . The aim of the present study was to determine if the therapeutic response to BT is enhanced by immediate and continuous activation of the injected muscles . Twenty SD patients were randomized into two groups following bilateral injections: vocal rest for 30 minutes and continuous vocalization for 30 minutes . Evaluations consisted of voice ratings by expert observers, acoustic measurements using computer analyses, and laryngeal aerodynamic measurements . The findings suggest that vocal rest, rather than vocalization, produces a superior and longer lasting response in SD patients receiving BT injections . It is recommended that SD patients refrain from post-injection vocalization to maximize the therapeutic effects of BT.

Neuropediatrics, 1995 Aug, 26(4), 214 - 6
Dystonic posture of lower extremities associated with myelomeningocele: successful treatment with botulinum A toxin in a six-month-old child; Heinen F et al.; We report on a six-month-old child with severe intermittent dystonic posture of both legs associated to a thoraco-lumbar myelomeningocele . The patient presented with a combination of progressive hypertonic knee extension and hip flexion . While the mobility of the right leg improved sufficiently after physiotherapy and splinting, satisfactory improvement of the left leg could be achieved only after local injections of botulinum A toxin, allowing for adequate functional motor development.

Prog Neurobiol, 1995 Aug, 46(5), 541 - 60
Effects of myotoxins on skeletal muscle fibers; Khan MA; This review highlights various aspects of a number of experimental myological alterations, induced by different chemical toxicants, including anticholinesterase, colchicine, vincristine, chloroquine, tetanus toxin, botulinum toxin, reserpine and emetine . Despite their chemical diversity and mechanism(s) of action, it is evident from the data discussed here that remarkably different toxic agents exert quite similar effects and induce toxic myopathies . The latter include preferential involvement of slow-twitch red muscle, mitochondrial derangement, denervation-like alterations, formation of membranous whorls, tubular aggregates, autophagic vacuoles and axonal sprouts . The non-invasive experimental models discussed here are valuable in studying various aspects of myopathology in the absence of any mechanical damage to the innervating elements from neurons to axonal terminals.

J Neurol, 1995 Aug, 242(8), 529 - 34
Botulinum toxin in cervical dystonia: low dosage with electromyographic guidance; Brans JW et al.; Sixty patients with idiopathic cervical dystonia were treated a total of 240 times with botulinum toxin type A (BTA) . Selected muscles were injected with BTA under electromyographic (EMG) guidance . The clinical effect was measured on the Tsui scale and a 10-point anchored visual analogue scale . A dosage of 150-300 mouse units was used in 77% of the treatments (mean 204 mouse units) . Based on the Tsui scale, 45% of 240 treatments were still effective at the moment of reinjection (median improvement 2 points) . Based on the 10-point anchored visual analogue scale, 73% of treatments were successful (median improvement 3 points) . Forty-eight patients (80%) responded favourably to the treatment . Side-effects were mild and transient . Dysphagia occurred in 9% of treatments . Antibody production was investigated in 41 patients and was negative in all . A striking difference from previous reports is the lower dosage used in this study . The clinical response, however, was similar to that of other studies . We conclude that a dosage of 200-400 mouse units BTA (Dysport) may also be effective in the treatment of cervical dystonia, but with fewer side effects . EMG guidance and application of BTA into deep cervical muscles may further improve the clinical effect.

J Neurol, 1995 Aug, 242(8), 504 - 7
Selective peripheral denervation for spasmodic torticollis: is the outcome predictable?
Braun V, Richter HP, Schroder JM.
If botulinum toxin fails in the treatment of cervical dystonia, selective peripheral denervation is now accepted as the best surgical option . Despite the very promising results, however, there is still a substantial group of patients who do not benefit from this procedure . Positive response to prior botulinum toxin therapy seems to be a very good predictor of outcome after selective peripheral denervation (P < 0.01) . The meaning of the histological findings of the resected nerves is uncertain . Patients with histologically proven pathological nerves do not seem to benefit more than patients with histological normal ones (P < 0.30).

Protein Sci, 1995 Aug, 4(8), 1490 - 7
Formation of ion channels in lipid bilayers by a peptide with the predicted transmembrane sequence of botulinum neurotoxin A; Oblatt-Montal M et al.; Synthetic peptides patterned after the predicted transmembrane sequence of botulinum toxin A were used as tools to identify an ion channel-forming motif . A peptide denoted BoTxATM, with the sequence GAVILLEFIPEIAI PVLGTFALV, forms cation-selective channels when reconstituted in planar lipid bilayers . As predicted, the self-assembled conductive oligomers express heterogeneous single-channel conductances . The most frequent openings exhibit single-channel conductance of 12 and 7 pS in 0.5 M NaCl, and 29 and 9 pS in 0.5 M KCl . In contrast, ion channels are not formed by a peptide of the same amino acid composition as BoTxATM with a scrambled sequence . Conformational energy calculations show that a bundle of four amphipathic alpha-helices is a plausible structural motif underlying the measured pore properties . These studies suggest that the identified module may play a functional role in the ion channel-forming activity of intact botulinum toxin A.

J Leukoc Biol, 1995 Aug, 58(2), 196 - 202
Nitric oxide stimulates ADP ribosylation of actin in association with the inhibition of actin polymerization in human neutrophils; Clancy R et al.; In these studies we provide conclusive evidence that (beta/gamma) actin present in human neutrophils is a substrate for nitric oxide (NO)-dependent ADP ribosylation and that this modification is associated with the inhibition of actin polymerization . A 43-kDa substrate for NO-dependent ADP ribosylation was identified as actin by four methods: (1) comigration with the botulinum C2 toxin substrate by two-dimensional gel electrophoresis (pI 5.2), (2) identity between the peptide map generated by V8 protease digestion of the NO and botulinum C2 substrates, (3) immunoprecipitation with antiactin antibodies, and (4) the ability of NO to ADP ribosylate purified neutrophil G-actin in the presence of plasma membrane cofactors . Because the ADP ribosylation of actin by the botulinum C2 toxin is known to inhibit F-actin polymerization, we examined the effect of NO on actin assembly . Flow cytometry revealed that NO inhibited formyl-methionine-leucine-phenylalanine (fMLP)-dependent (30 s at 37 degrees C) F-actin formation (108 +/- 8 vs . 89 +/- 6 relative fluorescence units, P < .02) . These results were confirmed by quantification of F-actin formation by gel scanning (10% sodium dodecyl sulfate gel, Coomassie, and densitometry): pretreatment of polymorphonuclear leukocytes with NO resulted in a reduction of fMLP-induced, cytoskeletal-associated F-actin, which was accompanied by an increase of Triton-soluble G-actin . NO also inhibited F-actin formation, as observed by means of rhodamine phalloidin staining of neutrophils adherent to a fibronectin-coated surface . This effect was accompanied by a dose-dependent inhibition of neutrophil adherence in NO-treated cells . The data indicate that NO inhibits cytoskeletal assembly and adherence in human neutrophils in association with the ADP ribosylation of actin.

Am J Gastroenterol, 1995 Aug, 90(8), 1319 - 21
Botulinum toxin for suspected pseudoachalasia; Vallera RA et al.; We describe a 74-yr-old man with stage III adenocarcinoma of the lung who presented with suspected malignancy-induced secondary achalasia and responded clinically to intrasphincteric injections of botulinum toxin type A (Botox, Allergen Inc., Irvine, CA) . We discuss the use of botulinum toxin in this setting, as well as diagnostic strategies to differentiate achalasia from pseudoachalasia.

Curr Opin Neurol, 1995 Aug, 8(4), 314 - 9
Essential tremor and its variants; Britton TC; Essential tremor is the commonest of movement disorders . Although sometimes prefaced with the term 'benign', it often causes significant disability . Diagnosis is based on the clinical finding of a postural tremor, predominantly affecting the upper limbs, that is absent at rest and not associated with extrapyramidal or cerebellar signs . There are, as yet, no specific anatomical, physiological, biochemical or genetic markers for the condition . Postural limb tremors, clinically indistinguishable from essential tremor, may occur in patients who have, or will later develop, other neurological conditions; whether such patients have essential tremor is a matter of controversy that will only be resolved with a better understanding of the pathophysiology of essential tremor . Positron emission tomography in patients with essential tremor reveals increased cerebellar activity even at rest, a finding that is consistent with the cerebellum having an important role in the generation of tremor . Abnormal cerebellar function has also been invoked to account for the abnormal manner in which patients with essential tremor perform rapid voluntary wrist movements . Molecular genetic studies in hereditary essential tremor have been initiated, but with negative results so far . Several new drug treatments have been tried, but with limited success; the role of thalamic stimulation and botulinum toxin in the treatment of essential tremor remains to be judged.

HNO, 1995 Aug, 43(8), 498 - 501
{Dystonia as the cause of pharyngolaryngeal motility disorders}; Zwirner P et al.; Dystonia as cause of pharyngo-laryngeal motility disorders has not been adequately considered in most clinical ENT practices . This case study of a patient with spasmodic torticollis, Meige's syndrome and pharyngo-laryngeal motility disorder was found to be due to dystonia as the underlying cause . The possibility of local symptomatic therapy with botulinum toxin injections has currently provided the physician with an effective means for alleviating the disorder.

No To Shinkei, 1995 Aug, 47(8), 749 - 54
{The clinical usefulness of botulinum toxin type A for spasmodic torticollis and facial spasm}; Mezaki T et al.; We studied the clinical effectiveness of botulinum toxin type A for spasmodic torticollis and that for facial spasm by multicenter, non-blinded study . The freeze-dried crystalline botulinum toxin type A (AGN 191622; Allergan Inc., Irvine, CA) was injected into the hyperactive muscles and the clinical course was followed for 22 weeks . Repeated injections were done, if necessary, with an interval of 4 weeks . The toxin was highly effective in both disorders . In spasmodic torticollis, clinical severity improved in 38 (63.3%) and the global improvement was seen in 39 (65.0%) out of 60 patients . Subjective improvement was seen in 56 (93.3%) . In facial spasm, 52 (92.9%) out of 56 patients improved after the treatment . Unfavorable reactions, mainly consisting of neck muscle weakness and dysphagia in torticollis and facial weakness in facial spasm, were mostly due to the excessive action of the toxin . They were usually mild and transient . No patients discontinued the trial because of side effects . Botulinum toxin injection is a very useful and safe method for the symptomatic treatment of spasmodic torticollis and facial spasm.

Biochem Biophys Res Commun, 1995 Jul 26, 212(3), 945 - 52
Poisoning by botulinum neurotoxin A does not inhibit formation or disassembly of the synaptosomal fusion complex; Otto H et al.; We investigated the effect of poisoning rat brain synaptosomes with botulinum neurotoxin A on the NSF-mediated disassembly of a complex consisting of syntaxin, SNAP-25 and synaptobrevin (fusion complex) . Botulinum neurotoxin A specifically removes 9 amino acids from the C-terminus of SNAP-25 and efficiently blocks KCl-evoked glutamate release from synaptosomes . We report that truncated SNAP-25 is incorporated into the fusion complex of poisoned synaptosomes . The presence of truncated SNAP-25 does not interfere with the NSF-induced disassembly of the fusion complex . Also, the release of truncated SNAP-25 from the fusion complex is similar to that of the native SNAP-25 . Since neither the formation of the complex nor its disassembly seems to be affected by the SNAP-25 fragment, this fragment is likely to block exocytosis by disrupting events between disassembly of the synaptosomal fusion complex and membrane fusion itself.

Lancet, 1995 Jul 15, 346(8968), 154 - 6
Pain and remote weakness in limbs injected with botulinum toxin A for writer's cramp; Sheean GL et al.; We describe two cases of a syndrome resembling neuralgic amyotrophy that occurred in limbs injected with botulinum toxin for writer's cramp . In both cases, one of the injections was followed by pain and the next, 7-10 weeks later, by weakness . We believe that unexplained pain in the shoulder region after an injection of botulinum toxin contraindicates a second injection.

FEBS Lett, 1995 Jul 3, 367(3), 246 - 50
Involvement of rho in GTP gamma S-induced enhancement of phosphorylation of 20 kDa myosin light chain in vascular smooth muscle cells: inhibition of phosphatase activity; Noda M et al.; In beta-escin-permeabilized cultured pig aortic smooth muscle cells GTP gamma S dose-dependently enhances Ca(2+)-induced, wortmannin-sensitive phosphorylation of 20 kDa myosin light chain (MLC20) . GTP gamma S does not potentiate thiophosphorylation of MLC20, but does inhibit its dephosphorylation . Pretreatment with C . botulinum exotoxin C3, which specifically ADP-ribosylates and inactivates the rho family of the small molecular weight G proteins, completely abolishes the effects of GTP gamma S . These results indicate that rho is involved in the GTP gamma S-induced enhancement of Ca(2+)-dependent MLC20 phosphorylation in aortic smooth muscle cells, and strongly suggest that this effect of rho is due to inhibition of protein phosphatase activity toward MLC20.

Neurol Neurochir Pol, 1995 Jul-Aug, 29(4), 481 - 8
{Own experience with botulinum treatment of dystonia}; Domzal T; Fifty-five patients were treated with botulin injections into the muscles showing dystonia, contracture or tremor . Twenty two of them had torticollis, 21 had blepharospasm, 10 had hemifacial spasm, and 2 had tremor . In all, 112 injections were done with good result in 64%, slight effect in 27% and without effect in 9% of the cases . Similar results have been reported from other centers in the world . Adverse effects were not significant and disappeared after several days or weeks . They included ptosis, speech and deglutition disturbances, general weakness and neurotic reactions . These adverse effects developed in 12 cases . In cases of tremor the dose as well as the technique of injections must be individualized . The method is an important therapeutic advance and can be applied in outpatient clinics.

Muscle Nerve, 1995 Jul, 18(7), 720 - 9
Five-year experience in the treatment of focal movement disorders with low-dose Dysport botulinum toxin; Van den Bergh P et al.; We report the results of botulinum toxin type A (Dysport, Porton Products, UK) treatment over 5 years in 107 patients with blepharospasm, Meige's syndrome, oromandibular dystonia, hemifacial spasm, cervical dystonia, and writer's cramp . Electromyography was used to localize dystonic muscles and guide Dysport injections in Meige's syndrome, oromandibular dystonia, cervical dystonia, and writer's cramp . All but 2 Meige's syndrome and 2 writer's cramp patients responded to treatment . Improvement was dramatic in blepharospasm (79%) and hemifacial spasm (90%); pronounced in cervical dystonia (74%); and moderate in Meige's syndrome (53%), oromandibular dystonia (57%), and writer's cramp (34%) . Although Dysport doses were 50-75% lower than usually reported, response and improvement rates as well as relapse intervals were similar to those of others . To treat cervical dystonia relapses, only 50% of the initial dose was required for continued optimal relief of symptoms . Low-dose Dysport was associated with a very low incidence of dysphagia in cervical dystonia.

Gastrointest Endosc Clin N Am, 1995 Jul, 5(3), 635 - 47
The use of high frequency endoscopic ultrasonography probes in the evaluation of achalasia; Miller LS et al.; Because its ultrasound beam cannot penetrate beyond a radius of 2 cm, HRES is limited in evaluating abnormalities within or adjacent to the esophageal wall . To date, however, it has provided new diagnostic capabilities in evaluating patients with several different esophageal disorders . EUS at lower frequencies has shown inconsistent results in evaluating the thickness of esophageal muscle layers in patients with achalasia . HRES has been able to more accurately measure the individual CSM, LSM, and TM layers of the muscularis propria and the mean measurements have been noted to increase in achalasia patients when compared with normals . HRES has been found to be clinically useful in assessing histologic damage following pneumatic dilatation and in localizing the LES during the administration of intrasphincter botulinum toxin injection in the treatment of achalasia.

Mov Disord, 1995 Jul, 10(4), 508 - 9
Auctioneer's jaw: a case of occupational oromandibular hemidystonia; Scolding NJ et al.; An auctioneer is described in whom focal dystonia of the jaw developed as an occupational symptom, occurring solely and predictably when he commenced his selling "patter" and resolving quickly on stopping . He responded well to treatment with intramuscular botulinus toxin.

Mov Disord, 1995 Jul, 10(4), 504 - 7
Hemimasticatory spasm in hemifacial atrophy: diagnostic and therapeutic aspects in two patients; Ebersbach G et al.; We report two cases of hemimasticatory spasm in association with progressive hemifacial atrophy . On the basis of neurophysiological and magnetic resonance imaging assessments, a peripheral irritation of the trigeminal nerve--probably due to entrapment of the motor branches in the infratemporal fossa--is suggested as the cause of the involuntary movement . Local injections of botulinum toxin type A into the masticatory muscles proved to be a successful treatment in both patients.

Mov Disord, 1995 Jul, 10(4), 466 - 71
Double-blind trial of botulinum toxin for treatment of focal hand dystonia; Cole R et al.; Ten patients with focal dystonia of the hand, all of whom had benefited in an open-label study of botulinum toxin, were treated with botulinum toxin-A in a double-blind study . Response was assessed by three measures: (a) subjective rating, provided by patients' reports of the effect of the injections on the dystonia; (b) objective testing, consisting of manual muscle testing (MRC scale) to measure muscle strength in all patients, timing of a writing sample and counting the number of errors of writing off-the-line in six patients with writer's cramp, counting the number of errors on a standard test of transcription in two patients with stenographer's cramp, and rating by professional musicians of the performances of two patients with musician's cramp; and (c) physicians' rating, provided by a review of the patients' videotaped performance by neurologists who were unaware of which treatment was administered . Eight of the 10 patients had greater subjective improvement with botulinum toxin than with placebo, and this impression was verified by at least one objective test in six patients . Two patients failed to have a better response to botulinum toxin than to placebo, and their reports were verified by the objective tests . This study confirms the efficacy of botulinum toxin in many patients with focal hand dystonia.

Mov Disord, 1995 Jul, 10(4), 455 - 9
Click-evoked vestibulocollic reflexes in torticollis; Colebatch JG et al.; A total of 26 patients with torticollis were studied using a recently developed technique for recording vestibulocollic reflexes from the sternocleidomastoid muscles in addition to conventional caloric tests of vestibular function . Previous reports of abnormalities of vestibulo-ocular reflexes in these patients were confirmed with just fewer than half having significant canal pareses or directional preponderances (nine of 20 tested) . In addition, there was a high incidence of abnormal click-evoked vestibulocollic reflexes (17 of 26 tested), which were not simply the result of prior treatment with botulinum toxin, nor due to unequal levels of muscle activation . In patients never previously treated with botulinum toxin (14 patients), the effect almost always consisted of suppressed responses in the sternocleidomastoid muscle ipsilateral to the direction of head turning . Because responses were not abnormal in all patients tested, and more commonly so in those with a history of torticollis of > or = 5 years (eight of nine patients) than in de novo patients, we suggest that the changes are more likely to be compensatory than causal.

Mov Disord, 1995 Jul, 10(4), 444 - 9
The blepharospasm disability scale: an instrument for the assessment of functional health in blepharospasm; Lindeboom R et al.; Assessment of the functional status in patients with blepharospasm is of major importance for clinical practice and outcome studies . The Blepharospasm Disability Scale (BDS) is specifically directed to measure the disability in these patients . The metric properties of this instrument were evaluated . Reliability, validity, and responsiveness to within-patient health changes over time of the BDS were assessed in 40 patients with essential blepharospasm treated with botulinum toxin injections . The reliability of the scale was sufficient for use on group level (Cronbach's alpha coefficient, 0.69) . Evidence of discriminant validity was provided by the difference in median score on the BDS between 21 newly admitted patients and 19 patients already under treatment (p < 0.001) . Convergent validity was supported by correlations between BDS and neurological impairment scores (range, Spearman correlation coefficients, 0.65-0.79) . Responsiveness to health changes was demonstrated by a significant difference between median BDS scores before treatment and 2 weeks after treatment with botulinum toxin (p < 0.01) . The BDS is a useful disease-specific instrument to assess disability . Completion of the questionnaire is easy and takes only a few minutes . The instrument is suitable for use in patient care, descriptive outcome studies, and should be considered in controlled clinical trials.

Ugeskr Laeger, 1995 Jun 26, 157(26), 3766 - 9
{Hemifacial spasms--clinical picture, diagnosis and treatment}; Jespersen JH et al.; A rectrospective study of 23 consecutive patients with hemifacial spasm is presented . Based on this experience, a practical approach to the diagnosis, investigation and treatment with injections of botulinum toxin is described . Magnetic resonance imaging angiography of the brain was performed in 20 patients an 15 controls . Contact between an artery from the vertebrobasilar circulation and the intracranial part of the facial nerve was observed ipsilaterally to the spasm in 17 patients (85%) and in two controls (7%) . Treatment with botulinum toxin was performed in 16 patients with moderate to good improvement in the majority of the patients . MR-angiography is recommended in order to exclude infrequent etiologies and as preoperative evaluation . Botulinum toxin injection is recommended as the symptomatic treatment of choice . The possibility of curative surgical intervention by microvascular decompression of the facial nerve is discussed.

J Biol Chem, 1995 Jun 16, 270(24), 14399 - 404
Endothelial caveolae have the molecular transport machinery for vesicle budding, docking, and fusion including VAMP, NSF, SNAP, annexins, and GTPases; Schnitzer JE et al.; Transport by discrete vesicular carriers is well established at least in part because of recent discoveries identifying key protein mediators of vesicle formation, docking, and fusion . A general mechanism sensitive to N-ethylmaleimide (NEM) is required for the transport of a divergent group of vesicular carriers in all eukaryotes . Many endothelia have an abundant population of non-coated plasmalemmal vesicles or caveolae, which have been reported with considerable controversy to function in transport . We recently have shown that like other vesicular transport systems, caveolae-mediated endocytosis and transcytosis are inhibited by NEM (Schnitzer, J . E., Allard, J., and Oh, P . (1995) Am . J . Physiol . 268, H48-H55) . Here, we continue this work by utilizing our recently developed method for purifying endothelial caveolae from rat lung tissue (Schnitzer, J . E., Oh, P., Jacobson, B . S., and Dvorak, A . M . (1995) Proc . Natl . Acad . Sci . U . S . A . 92, 1759-1763) to show that these caveolae contain key proteins known to mediate different aspects of vesicle formation, docking, and/or fusion including the vSNARE VAMP-2, monomeric and trimeric GTPases, annexins II and VI, and the NEM-sensitive fusion factor NSF along with its attachment protein SNAP . Like neuronal VAMPs, this endothelial VAMP is sensitive to cleavage by botulinum B and tetanus neurotoxins . Caveolae in endothelium are indeed like other carrier vesicles and contain similar NEM-sensitive molecular machinery for transport.

EMBO J, 1995 Jun 15, 14(12), 2723 - 30
VAMP-2 and cellubrevin are expressed in pancreatic beta-cells and are essential for Ca(2+)-but not for GTP gamma S-induced insulin secretion; Regazzi R et al.; VAMP proteins are important components of the machinery controlling docking and/or fusion of secretory vesicles with their target membrane . We investigated the expression of VAMP proteins in pancreatic beta-cells and their implication in the exocytosis of insulin . cDNA cloning revealed that VAMP-2 and cellubrevin, but not VAMP-1, are expressed in rat pancreatic islets and that their sequence is identical to that isolated from rat brain . Pancreatic beta-cells contain secretory granules that store and secrete insulin as well as synaptic-like microvesicles carrying gamma-aminobutyric acid . After subcellular fractionation on continuous sucrose gradients, VAMP-2 and cellubrevin were found to be associated with both types of secretory vesicle . The association of VAMP-2 with insulin-containing granules was confirmed by confocal microscopy of primary cultures of rat pancreatic beta-cells . Pretreatment of streptolysin-O permeabilized insulin-secreting cells with tetanus and botulinum B neurotoxins selectively cleaved VAMP-2 and cellubrevin and abolished Ca(2+)-induced insulin release (IC50 approximately 15 nM) . By contrast, the pretreatment with tetanus and botulinum B neurotoxins did not prevent GTP gamma S-stimulated insulin secretion . Taken together, our results show that pancreatic beta-cells express VAMP-2 and cellubrevin and that one or both of these proteins selectively control Ca(2+)-mediated insulin secretion.

JAMA, 1995 Jun 7, 273(21), 1710 - 2
Physical medicine and rehabilitation; Brandstater ME; An important priority has been the development of quantitative methods for measuring the disability status of patients undergoing rehabilitation . Intramuscular injection of botulinum toxin is an effective new therapy for spasticity due to a number of causes.

FEBS Lett, 1995 Jun 5, 366(1), 11 - 6
Lysophosphatidic acid-induced activation of protein Ser/Thr kinases in cultured rat 3Y1 fibroblasts . Possible involvement in rho p21-mediated signalling; Kumagai N et al.; Renaturation kinase assay was used to detect protein kinases activated by lysophosphatidic acid (LPA) in cultured rat 3Y1 fibroblasts . LPA activated several Ser/Thr protein kinases with apparent molecular weights of 145K, 85K, 64-65K (a doublet), and 60K (each named p145, p85, p64165 and p60, respectively) in addition to p43 mitogen activated protein (MAP)-kinase . Experiments using pertussis toxin and botulinum C3 exoenzyme showed that p145, p85, and p64165 kinases were activated by a pertussis toxin-insensitive rho p21-dependent pathway and that the activation of MAP-kinase was mediated by both the pertussis toxin-sensitive rho p21-independent and the pertussis toxin-insensitive rho p21-dependent pathways.

Laryngoscope, 1995 Jun, 105(6), 585 - 8
Treatment of vocal fold granuloma using botulinum toxin type A; Nasri S et al.; Contact granuloma of the vocal folds has been associated with abnormal use of the voice, and acid reflux may exacerbate the inflammatory process . Treatments have included voice therapy and antireflux measures . Surgical excision is considered in patients who do not respond to medical management . Localized injections of botulinum toxin type A (BOTOX) have been effective in patients with disorders of muscular control in the head and neck . In this study, granulomas resolved in six patients who underwent injection of the affected vocal folds . Botulinum toxin type A is probably successful because it prevents forceful closure of the arytenoids during phonation and coughing . Localized injection of this neurotoxin is promising both as an initial treatment and as an alternative treatment in patients who do not respond to standard therapy or who are poor surgical candidates.

Nervenarzt, 1995 Jun, 66(6), 465 - 7
{Increased residual urine volume after local injection of botulinum A toxin}; Schnider P et al.; After major head trauma, a 28-year-old male patient developed tetraparesis with marked left-sided contractions of the leg adductors . Spasticity was resistant to antispastic drugs and intensive physiotherapy . Therefore, we injected 12.5 ng botulinum A toxin (Dysport) in the left adductor longus and adductor magnus . Eight measurements of the post-micturition residual urine of the bladder before botulinum-A-toxin administration gave no evidence for urinary retention . Between day 5 and 14 after injection we measured pathologically increased urinary volumes up to 130 ml at five different points of time . This case report indicates possible subclinical side effects on the autonomic nervous system of the urinary bladder.

Nervenarzt, 1995 Jun, 66(6), 422 - 9
{Subgroups of torticollis spasmodicus from the psychosomatic viewpoint . Results of a cluster analysis of 144 cases . German Study Group of Dystonia Research}; Scheidt CE et al.; The question of subgroups in idiopathic spasmodic torticollis, which has been discussed in earlier studies in order to define etiologically heterogeneous patient populations has lost some of its relevance since with the injection of botulinum toxin an effective treatment is available . However, psychosocial distress is linked with spasmodic torticollis in a substantial number of patients . In order to define criteria for psychosocial interventions in addition to the treatment with botulinum toxin, a cluster analysis was carried out to identify high-risk populations in terms of psychological and social distress . Five subgroups were defined on the basis of eight variables . Two of these five groups, one group with rotational torticollis and one with laterocollis, emerged as particularly distressed by their physical complaints, the effects of their illness on various areas of life and in terms of psychological functioning . The consistency of the subgroups was tested and statistically confirmed by analysis of variance . In a cross-validation 83.02% of the ungrouped cases were predicted correctly . The authors suggest that the evaluation of psychological and social aspects of the condition should be part of the neurological assessment in order to offer appropriate support to patients, who reveal a high degree of psychological distress.

Nippon Ganka Gakkai Zasshi, 1995 Jun, 99(6), 663 - 8
{Botulinum toxin type A (BOTOX) for treatment of blepharospasm: an open label, dose-response study}; Iwashige H et al.; The clinical effect of Botulinum Toxin Type A (BOTOX) were studied in patients with blepharospasm . Clinical symptoms were evaluated with Jankovic's rating scale, widely used for blepharospasm . To see dose response, eyelid muscle force of the patients was measured with a device recently developed for measurement of eyelid muscle force . The results showed significant improvement (p = 0.0000) on Jankovic's rating scale in all patient groups after effective dose injections of 0.5, 1.25, and 2.5 U/site . Particularly the number of patients with marked improvement (decrease of six points or more in total score on Jankovic's rating scale) increased with higher dose injections . The eyelid muscle force also decreased by 33.2 +/- 28.1%, 41.7 +/- 25.1%, and 69.6 +/- 5.0% in patients groups after effective dose injections of 0.5, 1.25, and 2.5 U/site, respectively . The decrease of the eyelid muscle force showed significant dose response (p = 0.0254) . Mean duration of effect was 12.9 weeks in patients after effective dose injections of 1.25 U/site, which was significantly longer (p = 0.0205) than that of 9.6 weeks in patients after effective dose injections of 0.5 U/site . No severe adverse effects were observed . We concluded that BOTOX injections of 1.25 U/site or more are a safe and effective treatment of blepharospasm.

Brain, 1995 Jun, 118 ( Pt 3), 801 - 7
Physiological effects produced by botulinum toxin treatment of upper limb dystonia . Changes in reciprocal inhibition between forearm muscles; Priori A et al.; Patients with upper limb dystonia have abnormal reciprocal inhibition between flexor and extensor forearm muscles . To see whether botulinum toxin treatment alters segmental motor system function, we studied reciprocal inhibition between forearm flexor and extensor muscles, before and after botulinum toxin injection in forearm muscles in 12 patients with upper limb dystonia . Reciprocal inhibition was studied by conditioning the H reflex in forearm flexors with a radial nerve stimulus delivered at a range of time intervals . Botulinum toxin injection improved upper limb dystonia . Before botulinum toxin injection, the dystonic patients had a decreased second phase of reciprocal inhibition . After botulinum toxin injections this second abnormal phase of reciprocal inhibition increased . Botulinum toxin did not change the first phase of reciprocal inhibition . Botulinum toxin treatment also reduced the M wave and the H reflex by a similar amount but left the Hmax:Mmax ratio unchanged . Ample evidence has shown that the therapeutic effects of botulinum toxin in dystonia depend mainly on its neuromuscular junction blocking action . Our data now suggest a concurrent indirect effect on spinal cord circuitry, probably through the action of botulinum toxin on the intrafusal neuromuscular junction.

J Med Assoc Thai, 1995 Jun, 78(6), 281 - 8
Treatment of various movement disorders with botulinum A toxin injection: an experience of 900 patients; Poungvarin N et al.; A prospective open study of botulinum toxin A treatment for patients with various movement disorders at Siriraj Hospital, Mahidol University was analysed to evaluate its efficacy . The grand total of 900 patients comprised of a) 592 patients (65.78 per cent) with hemifacial spasm; b) 92 patients (10.22 per cent) with occupational cramp; c) 79 patients (8.78 per cent) with blepharospasm and Meige syndrome; d) 72 patients (8.00 per cent) with spasmodic torticollis; e) 19 patients (2.11 per cent) with hemidystonia and generalised dystonia; f) 11 patients (1.22 per cent) with spasmodic dysphonia; g) 10 patients (1.11 per cent) with spastic hemiparesis; and h) 25 patients (2.78 per cent) with miscellaneous group (i.e . tics, Gilles de la Tourette, facial myokimia, benign fasciculation, etc.) . The results of treatment for hemifacial spasm were classified as excellent in 486 patients (82.09 per cent), moderate improvement in 60 patients (10.14 per cent), mild improvement in 39 patients (6.59 per cent) and no improvement or worse in 7 patients (1.18 per cent) . There were complications of mild transient facial weakness in 50 patients (8.45 per cent) and mild ptosis in 12 patients (2.02 per cent) . The effect of botulinum toxin treatment lasted 3-6 months . In occupational cramp and spasmodic torticollis the good response rate was around two-thirds of all patients, whereas, blephalospasm, spasmodic dysphonia, spastic hemiparesis and tics responsed in 79-88 per cent of the patients . Botulinum toxin A injection is thus a simple, safe, and effective out-patient treatment for patients with various kinds of movement disorders but it is a costly therapy.

Cell, 1995 May 19, 81(4), 571 - 80
Different requirements for NSF, SNAP, and Rab proteins in apical and basolateral transport in MDCK cells; Ikonen E et al.; We used an in vitro system based on streptolysin O-permeabilized MDCK cells to study the involvement of NSF, SNAP, SNAREs, and Rab proteins in polarized membrane transport of epithelial cells . In MDCK cells, transport from the trans-Golgi network (TGN) to the basolateral plasma membrane is inhibited by anti-NSF antibodies and stimulated by alpha-SNAP . In contrast, transport from the TGN to the apical cell surface is not affected by anti-NSF antibodies or alpha-SNAP . Furthermore, apical transport is insensitive to Rab-GDI and tetanus and botulinum neurotoxins, which inhibit basolateral transport . These results provide evidence that the Rab-NSF-SNAP-SNARE mechanism operates in basolateral transport, while other molecules constitute the machinery for vesicular delivery in the apical pathway.

EMBO J, 1995 May 15, 14(10), 2317 - 25
Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro; Hayashi T et al.; The interaction of the presynaptic membrane proteins SNAP-25 and syntaxin with the synaptic vesicle protein synaptobrevin (VAMP) plays a key role in the regulated exocytosis of neurotransmitters . Clostridial neurotoxins, which proteolyze these polypeptides, are potent inhibitors of neurotransmission . The cytoplasmic domains of the three membrane proteins join into a tight SDS-resistant complex (Hayashi et al., 1994) . Here, we show that this reconstituted complex, as well as heterodimers composed of syntaxin and SNAP-25, can be disassembled by the concerted action of the N-ethylmaleimide-sensitive factor, NSF, and the soluble NSF attachment protein, alpha-SNAP . alpha-SNAP binds to predicted alpha-helical coiled-coil regions of syntaxin and SNAP-25, shown previously to be engaged in their direct interaction . Synaptobrevin, although incapable of binding alpha-SNAP individually, induced a third alpha-SNAP binding site when associated with syntaxin and SNAP-25 into heterotrimers . NSF released prebound alpha-SNAP from full-length syntaxin but not from a syntaxin derivative truncated at the N-terminus . Disassembly of complexes containing this syntaxin mutant was impaired, indicating a critical role for the N-terminal domain in the alpha-SNAP/NSF-mediated dissociation process . Complexes containing C-terminally deleted SNAP-25 derivatives, as generated by botulinal toxins type A and E, were dissociated more efficiently . In contrast, the N-terminal fragment generated from synaptobrevin by botulinal toxin type F produced an SDS-sensitive complex that was poorly dissociated.

Eur J Biochem, 1995 May 15, 230(1), 32 - 7
Autoregulation of actin synthesis in hepatocytes by transcriptional and posttranscriptional mechanisms; Reuner KH et al.; Treatment of rat hepatocytes with the filamentous-actin-stabilizing toxin phalloidin decreased the amount of globular actin by 77% in the cytosol and by 80% in the nucleus within 12 h . Simultaneously, actin mRNA was specifically increased by 230% . The de-novo synthesis of actin mRNA, as measured by nuclear run-on transcription, was enhanced by 250% . Treatment of cells with actinomycin D blocked the increase of actin mRNA . The apparent half-life of actin mRNA was not significantly altered during treatment with phalloidin . In contrast, the globular-actin-stabilizing botulinum C2 toxin increased the amount of cytosolic globular actin by 50% within 12 h . Simultaneously, the actin mRNA level was decreased by 62% . However, de-novo synthesis of actin mRNA was not impaired . The apparent half-life of actin mRNA was decreased by approximately 60% during treatment with C2 toxin . The data strongly suggest an autoregulatory control of actin synthesis on the basis of the globular/filamentous actin ratio in rat hepatocytes at the transcriptional as well as at the posttranscriptional levels.

Nebr Med J, 1995 May, 80(5), 109 - 15
Cervical dystonia: a review the role of botulinum toxin; Edwards LL et al.; Cervical dystonia, although rare in the general population, can severely affect the lives of those afflicted with the disease . Throughout history several theories have been proposed regarding its etiology and pathophysiology, from underlying mental disorders to post-infectious to altered basal ganglia and brainstem function . However, CD remains poorly understood . Because of its similarity to Idiopathic Torsion Dystonia a genetic basic is suspected, but is not proven . Without a true understanding of the disease treatment remains symptomatic, and begins with physical therapy and medications and progresses to consideration of surgery . These treatment strategies have provided some relief, which is usually less than satisfactory within a short period of time . Recently, the use of botulinum toxin has provided significant symptomatic relief of pain in CD and has been associated with subjective and objective improvement in head posture . This newest therapy, although symptomatic, restores a more normal head posture and pain relief enabling the individuals with CD to continue to be active and productive participants in life, providing a ray of hope to these people as we continue to search for a better understanding of the disease process and the development of more effective treatment strategies.

Neurology, 1995 May, 45(5), 897 - 902
Motor persistence of orbicularis oculi muscle in eyelid-opening disorders; Aramideh M et al.; We describe clinical and EMG findings in three patients with an inability to reopen the eyes after voluntary closure of the eyelids . Synchronous EMG recording from the levator palpebrae (LP) and orbicularis oculi (OrbOc) muscles revealed that after voluntary closure of the eyelids and upon the command to open the eyes, all three patients were unable to inhibit the "voluntary" contraction of the OrbOc muscles, while on clinical examination there was no evidence of ongoing OrbOc muscle contraction . This "motor persistence" was restricted predominantly to the pretarsal portion of the OrbOc . In one patient, it occurred as an isolated abnormality of the eyelid movement and was recorded as an additional EMG abnormality in two patients with blepharospasm and involuntary LP inhibition . Clinical examination alone cannot differentiate this type of disorder of supranuclear control of eyelid movement from involuntary LP inhibition; simultaneous EMG recording from the LP and OrbOc muscles is required . Injection of botulinum toxin into the pretarsal portion of OrbOc muscles is helpful.

Acta Otolaryngol, 1995 May, 115(3), 459 - 61
Frey's syndrome: treatment with botulinum toxin; Drobik C et al.; The effectiveness of botulinum toxin injections for the management of Frey's syndrome was studied . Botulinum toxin A (approximately 0.5 Units/cm2) was injected intracutaneously into the affected skin area as determined by Minor's starch iodine test . Gustatory sweating in the treated skin area ceased completely within 1 week and has not reappeared (12 months follow up until now in the first treated case) . There have been no side effects . It is concluded that local botulinum toxin injections are a highly effective and safe treatment for Frey's syndrome . Additional study is required to evaluate the duration of the therapeutic effect.

Mov Disord, 1995 May, 10(3), 333 - 6
"Off" painful dystonia in Parkinson's disease treated with botulinum toxin; Pacchetti C et al.; The "off" painful dystonia (OPD), usually concerning the feet, is a type of abnormal involuntary movement, induced by the chronic use of levodopa . It is mostly observed in the advanced stage of Parkinson's disease (PD), particularly in the early morning, in the evening, and late at night . Indeed, some patients have experienced OPD also during "on" periods when dystonic posture of the foot alternates with dyskinesia . The pain probably is due to sustained muscle contraction, which causes prolonged muscle spasm, as in primary dystonia or torticollis . Dopaminergic drugs like bromocriptine, pergolide, and especially apomorphine (s.c . infusions, or bolus), can dramatically improve the OPD . Anticholinergics baclofen and lithium are alos used in the management of OPD with some benefit . On the other hand, clinical experience shows that in many cases, these therapeutic procedures are not always enough to produce the expected results . Thirty PD patients (22 men and eight women) with OPD of the foot were treated with botulinum toxin (Botox, Btx) using electromyograms to guide injections . Dystonia was evaluated using a quantitative rating scale . The selection of the muscles for Btx treatment was carried out on the basis of foot posture . We injected Btx into tibialis posterior, tibialis anterior, gastrocnemius, flexor digitorum longus, and extensores hallucis longus with a median dose 40 IU for each muscle, distributed in two sites . In all patients, the pain improved within 10 days, whereas in 21 patients, the pain disappeared completely for 4 months (range, 3-7 months); a concomitant improvement in intensity of the dystonic spasm was also observed . No side effects were reported . Seven patients with associated "on" foot dystonia described an improvement of foot posture on walking . In conclusion, in this uncontrolled study, the use of Btx in OPD seemed a promising tool to improve pain linked to foot dystonia; however, because of the well-known underlying dopaminergic defect in OPD, the Btx therapy should be considered only if the dopaminergic treatment established for the management of OPD has failed.

Klin Oczna, 1995 May, 97(5), 147 - 51
{Principles and results of treatment in acquired paralysis of III, IV and VI nerves}; Kubatko-Zielinska A et al.; Between 1979-1994 120 patients with acquired paralysis of cranial nerves of ocular muscles were treated: 33 cases with paralysis of oculomotor nerve, 43 cases with paralysis of trochlear nerve, 44 cases with paralysis of abducens nerve . The majority of our patients were males (84-70.0%) aged 21-40 years (65-54.2%) . The paralysis was most frequently caused by traffic accidents (45 cases--37.5%) and assaults (26 cases--21.7%) . The most common symptom of paralysis was diplopia (109 patients--90.8%) . Only 12 persons (10.0%) were admitted during the first month of paralysis and visual disorders caused by it . In our group 18 patients (15.0%) were treated conservatively . Sixteen patients (13.3%) were given injections of botulin toxin A into the eye muscles . Surgical treatment, usually of several ocular muscles, was performed in 83 cases (67.5%) when diplopia was not reduced after 6-12 months . The method of surgery and results are presented.

J Pediatr Ophthalmol Strabismus, 1995 May-Jun, 32(3), 191 - 3
Botulinum A toxin treatment of a deviated orbital implant; Wortham E et al.; We report a case of progressive deviation of an implant that resulted in conjunctival thinning and an inadequately fit prosthesis . Such a case may have been treated with conjunctival grafting or replacement of the implant with an hydroxyapatite sphere . In this case, injection of botulinum A toxin (Botox) into the contracted medial rectus muscle contributed to the successful refitting of a new and stable prosthesis and improvement of the conjunctival integrity . Possible explanations of the progressive shift in the position of the implant and its postinjection stability are discussed.

Synapse, 1995 May, 20(1), 24 - 32
Synaptic transmission blockade increases plasminogen activator activity in mouse skeletal muscle poisoned with botulinum toxin type A; Tian WH et al.; Experimental denervation, either by nerve crush or axotomy, leads to a dramatic increase in muscle plasminogen activator (PA) activity, suggesting a regulation of muscle PA levels by some neural influence (Festoff et al., 1986, J . Cell Biol., 103:1415-1421; Hantai et al., 1990, Proc . Natl . Acad . Sci . U.S.A., 87:2926-2930) . The Botulinum toxin (BoTx) type A is known to selectively interrupt the release of acetylcholine without structurally altering synaptic morphology . In the present study we have used acute BoTx poisoning of hind limb muscles to further explore the neural regulation of muscle PA activities directly after poisoning and during the process of collateral reinnervation . Electromyographic recording and study of ultraterminal sprouting after zinc iodideosmium and silver-cholinesterase staining were used to monitor "denervation" and reinnervation . Muscle choline acetyltransferase activity did not decrease, as is observed after experimental denervation, but in contrast increased and, therefore, reflected the functional integrity of intramuscular nerve endings . Within 2 days of BoTx poisoning, muscle urokinase-PA, and to a lesser extent, tissue-PA activities, rose in muscle extracts as shown by an amidolytic assay and fibrin zymography . When reinnervation occurred, muscle urokinase-PA activity decreased but did not return to baseline levels within the 80 days of our study . These results suggest that cholinergic transmission-regulated events determine activity of muscle PAs and that PAs likely have a role in neuromuscular formation and plasticity.

Gastroenterol Nurs, 1995 May-Jun, 18(3), 92 - 5
Botulinum toxin: a new therapeutic use; Ogilvie J et al.; Botulinum Toxin is a powerful neurotoxin whose therapeutic uses for smooth muscle disorders have received little attention . Through innovative research, gastroenterologists at Johns Hopkins Hospital have developed a simple, less traumatic therapeutic use for this toxin in the treatment of achalasia . In this article, the authors describe the pathophysiology of this disease, its diagnosis and traditional treatment, and the newest potential therapy, Botulinum Toxin (BoTx) injection.

Prog Neurobiol, 1995 May, 46(1), 83 - 96
Molecular aspects of tetanus and botulinum neurotoxin poisoning; Ahnert-Hilger G et al.; Clostridial neurotoxins, tetanus and the botulinum toxins A-G, are high molecular weight proteins consisting of a heavy chain which is responsible for the internalisation and a light chain possessing a zinc-dependent proteolytic activity . They exclusively proteolyse either the vesicle membrane protein, synaptobrevin or two integral plasma membrane proteins, SNAP 25 and syntaxin . Together with cytosolic proteins these proteins form the SNARE complex involved in vesicle exocytosis, and their cleavage blocks the latter process . Clostridial neurotoxins have now become powerful tools to investigate the final events occurring during secretion in neuronal, endocrine, and non-neuronal cells . They are applied to dissect the specific interactions of the SNARE protein complex with cytosolic fusogens and other modulators of exocytosis . Whereas exocytosis is not essential for the survival of cells, the organism as a whole will fall victim to a few nanograms since interneuronal and neuromuscular transmission is vital to muscular control, especially in respiration . Although all clostridial neurotoxins by their light chains attack proteins of the SNARE complex, tetanus toxin and the various botulinum toxins differ dramatically in their clinical symptoms . The biological information for this difference resides on the respective heavy chains which select different transport routes carrying the light chain from the place of entrance to the final compartment of action . So far the different transport vesicles used either by the various botulinum neurotoxins or by tetanus toxin are not yet defined . Nevertheless at least one of the botulinum toxins serves as a beneficial drug in the treatment of severe neuromuscular spasms.

Neurosci Lett, 1995 Apr 28, 190(1), 33 - 6
Possible involvement of botulinum ADP-ribosyltransferase sensitive low molecular G-protein on 5-hydroxytryptamine (5-HT)-induced inositol phosphates formation in 5-HT2c cDNA transfected cells; Tohda M et al.; To clarify the involvement of botulinum ADP-ribosyltransferase sensitive low molecular G-proteins in 5-hydroxytryptamine (5-HT)-induced stimulation of phosphatidylinositol turnover, we examined the effects of 5-HT on inositol phosphates formation in COS 7 cells transfected with 5-HT2c receptor cDNA, but did not in non-transfected or vector-transfected cells . A typical 5-HT2c receptor antagonist mianserin (0.3-3 microM) inhibited the 5-HT-induced inositol phosphates formation . Treatment with botulinum toxin D preparation (20 micrograms/ml, 8 h) that contained botulinum C3 ADP-ribosyltransferase, blocked the 5-HT-induced inositol phosphate formation, although botulinum toxin A preparation that did not contain the enzyme did not have an influence . These results support our previous findings suggesting that low molecular weight G-proteins ADP-ribosylated by botulinum ADP-ribosyltransferase are involved in phospholipase C activity.

Biochem Biophys Res Commun, 1995 Apr 26, 209(3), 1102 - 10
Microfilament assembly is required for anti-IgM dependent MAPK and p90rsk activation in human B lymphocytes; Melamed I et al.; Mitogen-activated protein kinases (MAPK) are important mediators of signal transduction from the cell surface to the nucleus . These MAPK pathways serve different receptor-mediated signaling pathways leading to dual phosphorylation on serine/threonine and tyrosine residues . The mechanisms linking cytoplasmic MAPK activation to later events is still unclear . In this study we demonstrate that the microfilament system has an active role in MAPK activation . Cross-linking of surface IgM or direct activation of PKC with PMA resulted in time and concentration-dependent increases in F-actin content, MAPK (p42erk-2) activation, and phosphorylation of p90rsk . Pretreatment of the B cells with cytochalasin D or botulinum C2 toxin, microfilament-disrupting agents, prevented the increases in F-actin content as well as MAPK and p90rsk activation . These data indicate a role for the microfilament system in the complex and divergent functions of MAPK.

Biochemistry, 1995 Apr 25, 34(16), 5494 - 503
Blockade by botulinum neurotoxin B of catecholamine release from adrenochromaffin cells correlates with its cleavage of synaptobrevin and a homologue present on the granules; Foran P et al.; Botulinum neurotoxin type B blocks transmitter release via a selective endoproteolysis of the small clear vesicle membrane protein synaptobrevin that is essential for neuro-exocytosis . In view of the distinct characteristics of exocytosis of adrenochromaffin granules and considering the controversy over the presence of synaptobrevin on the latter, this study aimed to determine the molecular basis of the inhibition by this toxin of secretion from chromaffin cells . Thus, affinity-purified antibodies against a synaptobrevin synthetic peptide were used to quantify its concentrations in subcellular fractions of bovine adrenal medulla . The latter, as well as density gradient centrifugation and size-exclusion chromatography, showed that > 70% of the protein copurifies with the granules and their marker, dopamine beta-hydroxylase . Notably, much lower concentrations of synaptobrevin and synaptophysin were found in chromaffin granules than in synaptic small clear vesicles (approximately 9% and approximately 2%, respectively); however, isolated granule membranes exhibited greater enrichments (approximately 35% and approximately 9%) . A second immunoreactive protein was colocalized with synaptobrevin on chromaffin granules; in view of its susceptibility to the toxin and lower M(r), it is assumed to be cellubrevin and, also, because of its high homology . Involvement of synaptobrevin and cellubrevin in Ca(2+)-triggered granule exocytosis was established by the demonstrated correlation between the extent of botulinum neurotoxin B-induced inhibition of secretion and their selective proteolysis following introduction of the toxin into intact chromaffin cells . On the basis of these collective findings, it is concluded that these proteins occur on chromaffin granules and one or both are essential for exocytosis.

J Clin Gastroenterol, 1995 Apr, 20(3), 189 - 91
Endoscopic intrasphincteric injection of botulinum toxin for the treatment of achalasia; Rollan A et al.; Three patients with achalasia were treated with endoscopic injection of botulinum toxin (BoTx) . BoTx (80 U) was injected via a sclerotherapy needle into the lower esophageal sphincter (LES) . One patient complained of transient heartburn that resolved after omeprazole treatment . Two patients reported sustained symptomatic improvement . They were able to eat normally 48 h after treatment and have remained symptom free for 5 and 6.5 months, respectively . In these patients, esophageal manometry 4 months after treatment showed a marked reduction of resting LES pressure and the appearance of a previously absent LES relaxation after swallowing . The third patient had only a transient clinical improvement, with occasional dysphagia beginning 3 months after treatment . All patients showed unchanged aperistalsis of the esophageal body . Its less invasive nature compared with other therapeutic alternatives may give BoTX injection a role in the treatment of some patients with achalasia.

Brain, 1995 Apr, 118 ( Pt 2), 533 - 45
Botulinum toxin-induced myopathy in the rat; Hassan SM et al.; We have used histological and histochemical techniques at the light microscopical level, and electron microscopy to examine the myopathological changes in rat muscle up to 30 weeks following botulinum toxin injection . Apart from muscle fibre atrophy and related myofibrillar structural changes, the results show a number of striking abnormalities which developed and disappeared at different stages . During the first 4 weeks after toxin injection, vacuoles of variable size were seen in the sarcoplasm near myonuclei, both at and away from endplates . Following this, between 4 and 10 weeks post-injection, progressive degeneration of junctional folds and separation of some nerve terminals from the simplified postsynaptic membranes were observed . At different time points following recovery from the toxin-induced paralysis (evidenced by the increase in muscle fibre size and return of function) a number of abnormalities were still detectable in muscle fibres . These included the appearance within them of multiple arrays of sarcotubular profiles, focal areas lacking myofibrillar organization and mitochondria, abnormal mitochondrial aggregates showing crystalline inclusions, and extension of the postsynaptic densities along the full depth of junctional folds . Furthermore, targetoid-like areas were detected histochemically following recovery from the toxin-induced paralysis . The early extensive vacuolation of the sarcoplasm and the degeneration of junctional folds suggest a myotoxic effect of botulinum toxin . The late changes are likely to be (at least in part) related to the process of recovery following reinnervation.

Neurology, 1995 Apr, 45(4), 822 - 4
Botulinum toxin treatment of essential head tremor; Pahwa R et al.; We examined in a double-blind, placebo-controlled study the effects of botulinum toxin in 10 patients with essential head tremor . Each subject received two treatments approximately 3 months apart, one with botulinum toxin injections and another with normal saline injections into the sternocleidomastoid and splenius capitis muscles . The subjects were assessed before each treatment and at 2, 4, and 8 weeks after injections . There was moderate to marked improvement in clinical ratings in five subjects after botulinum toxin injections and in one subject after placebo . There was moderate to marked subjective improvement in five patients with botulinum toxin as compared with three subjects with placebo . Side effects were mild and transient . We conclude that botulinum toxin may be useful for patients with essential head tremor who have failed to benefit from oral medications.

Neurology, 1995 Apr, 45(4), 712 - 7
Botulinum toxin A for spasticity, muscle spasms, and rigidity; Grazko MA et al.; We studied the effects of botulinum toxin A in 12 patients with spasticity and in eight patients with rigidity . The study design was a double-blind, placebo-controlled crossover trial with botulinum toxin A versus saline . Using the Ashworth Scale for spasticity and the Unified Parkinson's Disease Rating Scale for rigidity, we gave the patients a tone grade before and 2 weeks after treatment . Improvement in tone by two grades or more was considered clinically significant . In the spasticity group, botulinum toxin A reduced the tone of all patients significantly, improved functionality and nursing care in eight of 12 patients, and alleviated painful spasms in five of five patients . In the rigidity group, muscle tone was decreased in seven of eight patients, functionality improved in four of seven, and joint and muscle pain decreased in four of five . We conclude that botulinum toxin A is effective against the disabling effects of spasticity and rigidity . The treatment was well tolerated.

Am J Ophthalmol, 1995 Apr, 119(4), 489 - 96
Unsatisfactory treatment of acquired nystagmus with retrobulbar injection of botulinum toxin; Tomsak RL et al.; PURPOSE: We quantified the effects of botulinum toxin injected into the retrobulbar space of patients with acquired nystagmus with prominent vertical or torsional components . METHODS: We measured binocular eye rotations in three planes before and after injection of botulinum toxin (10, 12.5, or 25 units) into the retrobulbar space of one eye of each of three patients, ages 28 to 37 years, with acquired pendular nystagmus . RESULTS: Retrobulbar injection of botulinum toxin abolished or reduced all components of the nystagmus in the treated eye in all three patients for about two to three months . The patient who received 25 units developed complete external ophthalmoplegia and blepharoptosis . The other two patients retained some voluntary movements but developed diplopia . In one patient, visual acuity improved from Jaeger 5 to Jaeger 1 . In a second patient, filamentary keratitis developed, and visual acuity declined from Jaeger 2 to Jaeger 7; keratitis was a recurrent problem one year after the botulinum toxin injection . In the third patient with predominantly torsional nystagmus, visual acuity was unchanged at Jaeger 2 . No patient was pleased with the results, because of blepharoptosis, diplopia, or discomfort (from keratitis), and none elected to repeat the procedure . CONCLUSIONS: The side effects of botulinum toxin administered by retrobulbar injection limit its therapeutic value in the treatment of acquired nystagmus . Even smaller doses that do not abolish nystagmus may produce troublesome diplopia.

Otolaryngol Head Neck Surg, 1995 Apr, 112(4), 566 - 71
Rhinorrhea is decreased in dogs after nasal application of botulinum toxin; Shaari CM et al.; At this time no effective long-term therapy exists for the excessive secretion of vasomotor rhinitis . Because rhinorrhea is under parasympathetic control, it was theorized that botulinum toxin--a powerful and long-acting cholinergic blocker that has been successful in the treatment of dystonia--might be useful in blocking the cholinergic control of rhinorrhea . Four male mongrel dogs were studied . Fifty units of type A botulinum toxin was soaked into sterile gauze, which was then packed into the left nasal cavity of each dog for 1 hour . Saline-soaked gauze was similarly introduced into the right nasal cavity to serve as control . Six days later, rhinorrhea was produced by inserting a bipolar needle electrode into the sphenopalatine ganglion and electrically stimulating for 10 minutes (6 mA, 50 Hz) . Nasal secretions were collected with a suction catheter placed in the nasal vestibule . Three of four dogs exposed to the toxin showed a 41% average decrease in rhinorrhea (specifically 53%, 41%, and 30%) . One dog showed a 10% increase in secretion after exposure to the toxin . We conclude that topically applied botulinum toxin reduced neurally evoked rhinorrhea by an average of 41% . Because some secretion is mediated by noncholinergic neurotransmitters such as vasoactive intestinal peptide, topical application of an anticholinergic substance has limitations . However, because all the nasal parasympathetic nerves appear to originate from cholinergic synapses in the sphenopalatine ganglion, direct injections of toxin into this ganglion may possibly allow complete blockade of all cholinergically mediated rhinorrhea.

Neuroscience, 1995 Apr, 65(3), 905 - 15
Impairment of syntaxin by botulinum neurotoxin C1 or antibodies inhibits acetylcholine release but not Ca2+ channel activity; Mochida S et al.; The involvement of syntaxin, an omega-conotoxin-sensitive Ca2+ channel-associated protein, in acetylcholine release was studied at synapses formed between rat sympathetic neurons in culture . Transmission at these synapses involved omega-conotoxin-sensitive Ca2+ channels because a dose-dependent inhibition was observed when omega-conotoxin was bath-applied . Confocal microscope examination of immunofluorescent staining showed that syntaxin had a similar distribution to synaptic vesicle-associated membrane proteins, synaptophysin and vesicle-associated membrane protein/synaptobrevin-2, indicating that syntaxin molecules are concentrated in the presynaptic terminals . Botulinum neurotoxin C1 applied extracellularly or intracellularly into presynaptic neurons blocked synaptic transmission . Introduction of a monoclonal antibody, or polyclonal antibodies, to syntaxin into the presynaptic neuron depressed the evoked release of acetylcholine without affecting Ca2+ influx through Ca2+ channels . These results suggest that syntaxin plays an important role in release of neurotransmitter by a nerve impulse and that this mechanism is downstream of Ca2+ influx.

J Formos Med Assoc, 1995 Apr, 94(4), 189 - 92
Double-blind, placebo-controlled study of botulinum toxin injections in the treatment of cervical dystonia; Lu CS et al.; Botulinum toxin injections were given to 23 patients with cervical dystonia in a double-blind, placebo-controlled, crossover study . Each patient was randomly given 100 units of botulinum A toxin and placebo in two respective sets of injections into the two most active contracting neck muscles at least 3 months apart . Before and 3 weeks after each set of injections, the cervical dystonia was "blindly" scored by the investigator . Each patient was also asked to independently score the improvement of the dystonia and neck pain . Four patients dropped out and one patient showed spontaneous remission . The remaining 18 patients completed the study and showed a significant improvement after the injections of botulinum toxin in comparison with that of the placebo . The effect started about 1 week after injection and the maximal effect lasted about 11 weeks in patients with a satisfactory response . The neck pain was satisfactorily relieved in 9 of 10 patients affected . There were no significant side effects . Thus, botulinum toxin injection is effective in treating cervical dystonias; injection of a higher dose of botulinum toxin in a greater number of neck muscles is required for those patients with complex cervical dystonia.

Toxicon, 1995 Apr, 33(4), 559 - 67
Structural predictions of the channel-forming region of botulinum neurotoxin heavy chain; Lebeda FJ et al.; A novel combination of theoretical approaches was exploited to predict which amino acid residues of various botulinum neurotoxin serotypes participate in forming ion channels . Estimates of sequence hydrophobic moments were used initially to identify residues within amphipathic regions in the N-terminal half of the heavy chain . A neural network algorithm was then used to make additional secondary structural predictions for these regions . Together, these approaches predicted a complimentary pattern of four, adjacent amphipathic, possibly transmembrane, regions that may be separated by solvent-exposed loops . Both the hydrophobic moment and the neural net analyses predicted that at least one of these amphipathic segments may be in an extended conformation . These theoretical results are discussed in view of our current knowledge of other transmembrane structures.

Toxicon, 1995 Apr, 33(4), 551 - 7
A study of zinc-dependent metalloendopeptidase inhibitors as pharmacological antagonists in botulinum neurotoxin poisoning; Deshpande SS et al.; Zinc-dependent metalloprotease inhibitors phosphoramidon, captopril and a peptide hydroxamate were studied as potential pretreatment compounds by examining their ability to delay the onset or to prolong the time to 50% block of nerve-elicited muscle twitch tension in the mouse phrenic-nerve diaphragm (in vitro at 36 degrees C) after botulinum neurotoxin serotypes A and B (BoNT-A, BoNT-B) . Addition of BoNT-A or BoNT-B (1 x 10(-10) M) produced 50% block of the twitch response at 56 +/- 9 min and 76 +/- 4 min, respectively . Preincubation (45 min) of muscles with phosphoramidon (0.2 mM) prolonged the time to 50% block by 15 min in BoNT-B-poisoned muscles with no effect on the time-course of paralysis in BoNT-A exposed muscles . When the same quantities of BoNT-A or BoNT-B (equivalent to 1 x 10(-10) M bath concentration) were preincubated for 2 hr with phosphoramidon (equivalent to 0.2 mM final bath concentration), and the incubation mixture was added to the muscle chamber, the times to 50% block were prolonged by 38 min and 18 min for BoNT-B and BoNT-A, respectively . Preincubation of diaphragms with captopril (up to 10 mM) or peptide hydroxamate (75 microM) failed to antagonize BoNT-A or BoNT-B-induced neuromuscular block . Among the three metalloprotease inhibitors examined here, only phosphoramidon showed a significant protection against both serotypes of BoNT . A search for better inhibitor compounds specifically tailored to match the active site on BoNT molecule deserves attention.

Toxicon, 1995 Apr, 33(4), 539 - 49
Interactions between heavy metal chelators and botulinum neurotoxins at the mouse neuromuscular junction; Sheridan RE et al.; Exposure of isolated mouse hemidiaphragms to botulinum neurotoxins, 0.1 nM BoNT-A or BoNT-B, at 36 degrees C reduced nerve-elicited peak isometric twitch tension to 50% of control values at 55 min (BoNT-A) to 68 min (BoNT-B) after application . Either coincubation of BoNT with the heavy metal chelator TPEN, preincubation with TPEN followed by BoNT, or application of TPEN after BoNT but before neuromuscular block, delayed the onset of muscle failure in a dose-dependent manner by up to five-fold . TPEN doses between 2 and 10 microM were required to antagonize significantly the muscle block produced by BoNT, and the delay in onset was maximal between 10 and 50 microM TPEN . Treatment of muscles with a Zn(2+)-TPEN coordination complex, rather than TPEN alone, eliminated any beneficial effects of TPEN on BoNT intoxication, indicating that these effects were mediated by chelation of Zn2+ . Other metal chelators that were not as membrane permeant as TPEN were ineffective in delaying BoNT paralysis, suggesting that TPEN acts by chelating intraterminal Zn2+ . In the absence of BoNT, TPEN caused a dose-dependent increase in nerve-elicited twitch tension with a half-maximal concentration at 8 microM . There was no corresponding change in twitches from direct electrical stimulation of the muscle . After BoNT (A or B serotype) had reduced the muscle twitch by 20 to 70%, however, subsequent application of TPEN rapidly depressed nerve-elicited twitches . The shift from potentiation to depression after BoNT treatment suggests that presynaptic vesicle mobilization and/or release involve Zn(2+)-dependent enzymes and that BoNTs interact with these enzyme pathways.

Toxicon, 1995 Apr, 33(4), 527 - 37
Antagonism of botulinum toxin-induced muscle weakness by 3,4-diaminopyridine in rat phrenic nerve-hemidiaphragm preparations; Adler M et al.; The effects of the potassium channel inhibitor and putative botulinum toxin antagonist 3,4-diaminopyridine (3,4-DAP) were investigated in vitro on the contractile properties of rat diaphragm muscle . In the presence of 100 pM botulinum neurotoxin A (BoNT/A), twitches elicited by supramaximal nerve stimulation (0.1 Hz) were reduced to approximately 10% of control in 3 hr at 37 degrees C . Addition of 3,4-DAP led to a rapid reversal of the BoNT/A-induced depression of twitch tension . In the presence of 100 microM 3,4-DAP, antagonism of the BoNT/A-induced blockade began within 30-40 sec and reached 82% of control with a half-time of 6.7 min . The beneficial effect of 3,4-DAP was well maintained and underwent little or no decrement relative to control for at least 8 hr after addition . Application of 1 microM neostigmine 1 hr after 3,4-DAP led to a further potentiation of twitch tension, but this action lasted for < 20 min . Moreover, neostigmine caused tetanic fade during repetitive stimulation . In contrast to the efficacy of the parent compound, the quaternary derivative of 3,4-DAP, 3,4-diamino-1-methyl pyridinium produced little or no twitch potentiation up to a concentration of 1 mM . The potassium channel blocker, tetraethylammonium, generated a transient potentiation followed by a sustained depression of twitch tensions . It is concluded that 3,4-DAP is of benefit in antagonizing the muscle paralysis following exposure to BoNT/A . Co-application of neostigmine or tetraethylammonium with 3,4-DAP, however, appears to confer no additional benefit.

Toxicon, 1995 Apr, 33(4), 507 - 14
Characterization of a rabbit serum raised against a botulinum toxin type A binding protein from presynaptic plasma membranes from Torpedo electric organ; Canals JM et al.; Botulinum neurotoxin type A blocks acetylcholine release from the peripheral nervous system . We have previously described a putative botulinum neurotoxin type A receptor of presynaptic plasma membranes from Torpedo . The electric organ of Torpedo, which is largely enriched in cholinergic nerve endings, is homologous to the neuromuscular junction, allowing us to isolate large scale of presynaptic components . In order to characterize this protein we have raised a polyclonal antibody (a-P140) against this receptor . The antiserum a-P140 recognizes a 140,000 mol . wt band in non-reducing conditions and an 80,000 band in reducing conditions . The immunohistochemistry assay reveals the P140 protein on the ventral face of the electrocytes where the nerve terminals are localized . Moreover, a-P140 antiserum recognizes the P140-BoNT/A complex after binding and cross-linking experiments . In addition, we have immunoprecipitated an in vitro translated product which is closely coincident in mol . wt to the 80,000 band of the receptor.

N Engl J Med, 1995 Mar 23, 332(12), 774 - 8
Intrasphincteric botulinum toxin for the treatment of achalasia; Pasricha PJ et al.; BACKGROUND . Achalasia is a disorder of swallowing in which the lower esophageal sphincter fails to relax . We report the use of botulinum toxin, a paralytic agent, for the treatment of this condition . METHODS . In a double-blind trial, 21 patients with achalasia received either 80 units of botulinum toxin or placebo, injected endoscopically into the lower esophageal sphincter . One week later, the response to treatment was assessed on the basis of changes in the symptom scores (measured on a scale from 0 to 9), pharyngoesophagograms, and results of esophageal manometric and scintigraphic studies . Patients who received placebo initially were subsequently treated with botulinum toxin . After six months, esophageal scintigraphy was repeated . RESULTS . One week after treatment, the mean decrease in the symptom score was 5.4 points for the patients treated with botulinum toxin and 0.5 point for the placebo group (P = 0.001) . The mean decrease in the pressure of the lower esophageal sphincter was 33 percent in the treatment group, as compared with a mean increase of 12 percent in the placebo group (P = 0.02), and the mean increase in the width of the opening of the lower esophageal sphincter was 204 percent in the treatment group, as compared with a mean decrease of 14 percent in the placebo group (P = 0.02) . Nineteen of the 21 patients treated with botulinum toxin had symptomatic improvement initially; after six months 14 patients were still in remission . This improvement was accompanied by a decrease in esophageal retention that was sustained at six months (46 percent, as compared with a pretreatment value of 77 percent; P = 0.04) . There were no serious adverse effects . CONCLUSIONS . Injection of botulinum toxin into the lower esophageal sphincter is an effective, safe, and simple method of treatment for achalasia, with results that are sustained for several months.

Neurosci Lett, 1995 Mar 10, 187(3), 161 - 4
Effects of local injections of botulinum toxin on electrophysiological parameters in patients with hemifacial spasm: role of synaptic activity and size of motor units; Glocker FX et al.; Ten patients with typical hemifacial spasm were examined before and after treatment with local injections of botulinum toxin type A . After a mean follow-up period of 38 days there was a reduction of the compound muscle action potential (CMAP) of the injected orbicularis oculi muscle of 40% . Ephaptic transmission studied by selective stimulation of facial nerve branches revealed a preserved delayed response of the affected mentalis muscle . However, no delayed response could be recorded in the injected orbicularis oculi muscle in nine patients . The discrepancy between complete loss of the delayed (ephaptic) response and only moderate reduction of the CMAP amplitude of the direct response may be explained by preferential uptake of botulinum toxin type A by hyperactive synapses involved in ephaptic transmission.

Neuroreport, 1995 Mar 7, 6(4), 637 - 41
Calcium-dependent endogenous proteolysis of the vesicle proteins synaptobrevin and synaptotagmin; Hausinger A et al.; The synaptic vesicle integral protein synaptobrevin/VAMP is a target of the clostridial metalloproteases tetanus toxin and botulinum toxins . We provide evidence that synaptobrevin can also be cleaved by an endogenous protease . As revealed by Western blotting proteolysis is calcium-dependent, results in the formation of an 8 kD peptide that becomes apparent within 10 min . Proteolysis can be inhibited by the chelating agents EGTA and EDTA, whereas other protease inhibitors failed to prevent degradation . In addition, a proteolytic degradation of the synaptic vesicle specific protein synaptotagmin could be observed . Other proteins including the synaptic vesicle proteins synapsin I and synaptophysin remained unaltered . Partial calcium-dependent degradation of select synaptic vesicle proteins may play a role in the life cycle of the organelle.

Neurology, 1995 Mar, 45(3 Pt 1), 506 - 8
Comparison of therapeutic efficacies of type A and F botulinum toxins for blepharospasm: a double-blind, controlled study; Mezaki T et al.; Type F botulinum toxin can be used for treating patients with dystonia who become refractory to type A toxin injection due to antibody development . We compared the therapeutic efficacy of type F botulinum toxin to that of type A toxin in a self-controlled, double-blind clinical trial . In nine patients with blepharospasm, we injected type A toxin on one side and the same units of type F toxin on the other side . Although the onset of clinical effect, maximal benefit, and adverse reactions were similar between type A and F toxins, the duration of the clinical effect was significantly shorter on the side injected with type F toxin . Although type F toxin proved its promise as an alternative to type A toxin, its usefulness is limited by the shorter duration of action.

J Cell Biol, 1995 Mar, 128(6), 1019 - 28
SNAP-25 is expressed in islets of Langerhans and is involved in insulin release; Sadoul K et al.; SNAP-25 is known as a neuron specific molecule involved in the fusion of small synaptic vesicles with the presynaptic plasma membrane . By immunolocalization and Western blot analysis, it is now shown that SNAP-25 is also expressed in pancreatic endocrine cells . Botulinum neurotoxins (BoNT) A and E were used to study the role of SNAP-25 in insulin secretion . These neurotoxins inhibit transmitter release by cleaving SNAP-25 in neurons . Cells from a pancreatic B cell line (HIT) and primary rat islet cells were permeabilized with streptolysin-O to allow toxin entry . SNAP-25 was cleaved by BoNT/A and BoNT/E, resulting in a molecular mass shift of approximately 1 and 3 kD, respectively . Cleavage was accompanied by an inhibition of Ca(++)-stimulated insulin release in both cell types . In HIT cells, a concentration of 30-40 nM BoNT/E gave maximal inhibition of stimulated insulin secretion of approximately 60%, coinciding with essentially complete cleavage of SNAP-25 . Half maximal effects in terms of cleavage and inhibition of insulin release were obtained at a concentration of 5-10 nM . The A type toxin showed maximal and half-maximal effects at concentrations of 4 and 2 nM, respectively . In conclusion, the results suggest a role for SNAP-25 in fusion of dense core secretory granules with the plasma membrane in an endocrine cell type- the pancreatic B cell.

Am J Ophthalmol, 1995 Mar, 119(3), 376 - 7
Hemifacial spasm and osteitis deformans; Ing EB et al.; PURPOSE/METHODS: A patient with osteitis deformans (Paget's disease) and hemifacial spasm underwent magnetic resonance tomographic angiography . Bone-modulating bisphosphonates and botulinum injection were administered to treat the hemifacial spasm . RESULTS/CONCLUSIONS: Computed tomography showed marked temporal bone overgrowth . Magnetic resonance tomographic angiography showed no vascular compression of the facial nerve root . The hemifacial spasm failed to resolve with intravenous pamidronate . Subsequent botulinum injection rendered the patient spasm free for 22 weeks . Further research on the use of bisphosphonates in the treatment of pagetoid hemifacial spasm is required.

J Exp Med, 1995 Mar 1, 181(3), 1071 - 9
Nerve growth factor triggers microfilament assembly and paxillin phosphorylation in human B lymphocytes; Melamed I et al.; Increasing evidence suggests that the nervous system is involved in allergic inflammation . One of the potential regulatory molecules of the neuroimmune system is nerve growth factor (NGF) . Recent studies from our group demonstrated the presence of a functional NGF receptor (NGFR) on human B lymphocytes . Moreover, we showed that gp140trk tyrosine kinase, which serves as an NGFR, was involved in transduction of early signaling events in human B lymphocytes . The mechanisms by which NGF initiates the signaling cascade and the link between the neuroimmune systems are unknown . We have focused on the role of the cytoskeleton as a possible mediator for transduction of signals induced by NGF . Polymerized actin (F-actin) content was determined by fluorescent staining and immunoblotting with antiactin antibody . Addition of NGF caused a time- and concentration-dependent increase in F-actin content, and maximum effects were noted after 1 min . These increases in F-actin content and NGF-induced thymidine incorporation could be blocked by incubating the cells with cytochalasin D and botulinum C2 toxin before the addition of NGF . Incubation of human B lymphocytes with 10 nM K252a, an inhibitor of Trk kinase, decreased NGF-induced microfilament assembly by 75% . In immunoprecipitation experiments, addition of NGF to B cells induced a rapid increase in the tyrosine phosphorylation of paxillin, one of a group of focal adhesion proteins involved in linking actin filaments to the plasma membrane . Coimmunoprecipitation studies demonstrated the association between gp140trk kinase and paxillin . Together, these observations suggest that actin assembly is involved in NGF signaling in human B cells, and that paxillin may be essential in this pathway after phosphorylation by gp140trk kinase.

Fortschr Neurol Psychiatr, 1995 Mar, 63(3), 99 - 105
{Clinical aspects and therapy of dystonias}; Huber M; Dystonia is characterized by slow, repetitive, involuntary, often twisting movements leading to sustained abnormal postures and can be focal or generalised . In most cases, etiology remains unrevealed, some cases are hereditary, others have metabolic origin or can be attributed to focal brain lesions . The most common types of dystonia, spasmodic torticollis and blepharospasm, can now be treated successfully, albeit symptomatically, with Botulinum toxin injections into the affected muscles . A practical guide to differential diagnostic and therapeutic strategies is presented.

J Physiol, 1995 Mar 1, 483 ( Pt 2), 397 - 406
Number of junctional acetylcholine receptors: control by neural and muscular influences in the rat; Andreose JS et al.; 1 . The number of acetylcholine receptors (AChRs) per neuromuscular junction in soleus muscles of adult rats was estimated from counts of 125I-alpha-bungarotoxin binding sites . The muscles were either denervated, denervated and electrically stimulated, paralysed by botulinum toxin (BoTX), or paralysed by tetrodotoxin (TTX) . 2 . After denervation, the number of junctional AChRs was normal after 18 days and then fell to 54 and 35% of normal after 33 and 57 days, respectively . 3 . Direct high frequency muscle stimulation (100 Hz) maintained a normal number of junctional AChRs for at least 2 months when the stimulation started on the day of denervation . When the stimulation was started progressively later, the effect of the stimulation on AChR number disappeared within about a week . The disappearance was gradual and appeared to affect all the muscle fibres equally . 4 . Stimulation at 100 Hz, starting on the day of denervation and stopping after 18 days, did not prevent the endplates from losing AChRs during the subsequent 15 days without stimulation . Thus 100 Hz stimulation and innervation are not equivalent in their effects on junctional AChR number . 5 . Direct low frequency muscle stimulation from the day of denervation did not maintain a normal number of junctional AChRs, as the number of AChRs fell to 70 and 62% of normal after 33 days of stimulation at 20 and 10 Hz, respectively . 6 . Endplates paralysed by BoTX or TTX for 33 days lost about as many junctional AChRs (54 and 55%) as endplates denervated for 33 days (46%) . Direct stimulation at 100 Hz during the last 15 days of BoTX treatment reduced but did not prevent this AChR loss (36% loss at 33 days) . 7 . The results show that when motor nerve terminals in rat soleus muscles are removed by axotomy, they leave a 'trace' which, in conjunction with appropriate muscle stimulation, can maintain a normal number of AChRs in the postsynaptic region . In non-stimulated muscles the trace responsible for this maintenance disappears within about a week . In stimulated muscles it persists for at least 2 months . From indirect evidence it appears that the trace is a factor, or the postsynaptic effect of a factor, released by impulse activity in the nerve, and that its degradation after denervation is accelerated by the acute effects of nerve degeneration.

Rinsho Shinkeigaku, 1995 Mar, 35(3), 251 - 5
{Treatment of spastic paraparesis with botulinum toxin with reference to beneficial effects, disease severity and long-term treatment}; Takenaga S et al.; We administered local botulinum toxin injections on the leg adductors of 12 patients with spastic paraparesis (9 patients with HAM, 2 patients with spinal spastic paraparesis, 1 patient with an identified degenerative disease) . Two of them were wheelchair-bound and the other patients could walk with or without help . The patients were assessed by the time to walk 10 m and the spasticity score which was derived from the degree of muscle tone and spasm frequency of leg adductors . After the initial injection, 7 of the 12 patients improved spasticity scores and 8 of the 10 patients could walk 10 m within a shorter time . The time to walk 10 m was markedly shortened in moderate cases . However, one patient complained of leg weakness and the time to walk 10 m was prolonged . Five of the 12 patients received injections 3 to 7 times, and were followed up for a mean of 16.2 months . In 4 of the 5 patients, repeated injections could maintain the improvement of spasticity score and time to walk 10 m . However, injection was discontinued in one patient because of leg weakness . The other side effects were pain and swelling at the injected site and dysarthria . However, these side effects were slight and transient and did not require treatment . No other systemic side effects were observed . In conclusion, the beneficial effects of botulinum injections to spastic paraparesis were (1) improvement of objective symptoms in mild cases, (2) improvement of ADL in moderate cases, and (3) improvement of objective symptoms and ease of nursing care in severe cases . Furthermore, we confirmed the long-term efficacy and safety of botulinum toxin.

Laryngoscope, 1995 Feb, 105(2), 144 - 8
The effect of botulinum toxin type A injection on compound muscle action potential in an in vivo rat model; Cichon JV Jr et al.; Serial measurements were performed on the compound muscle action potential (CMAP) amplitude and the force generated by the rat lower hind limb flexors to investigate the time course of intramuscular injections of botulinum toxin type A (BOTOX) . Thirty animals were used in this in vivo rat model . CMAP amplitude and muscle force were measured at predetermined intervals for 28 weeks . Compound muscle action potential amplitude and force declined markedly the first 5 to 7 days after injection of BOTOX but recovered in a near linear manner . The response magnitude and recovery rate were dose-dependent . Recovery of CMAP amplitude preceded recovery of muscle force . No clear evidence of a systemic effect on the untreated leg or a concentration effect was found . CMAP amplitude may be useful in determining optimal timing of repeat injections in treating neuromuscular disorders.

J Neurol Neurosurg Psychiatry, 1995 Feb, 58(2), 232 - 5
Treatment of chronic limb spasticity with botulinum toxin A; Dunne JW et al.; The purpose of this open study was to find out whether botulinum toxin A (BTX-A) relieves the signs and symptoms of chronic limb spasticity . The study comprised 40 patients, aged 12-82 years, with moderate to severe spasticity of the upper (13) or lower limbs (27) refractory to conventional physical and medical treatments . Outcome measures were clinical and blinded videotape assessments of spasticity and motor function . Electromyography guided BTX-A injections were given in one or two sessions at total doses averaging 175 U in the upper limb (range 70-270 U) and 221 U in the lower limb (range 100-500 U) . Thirty four patients (85%) derived worthwhile benefit, with improved limb posture and increased range of passive motion in 31, pain reduction in 28 of 31 with pain, and improved function in 16 . Side effects were limited to local and usually mild discomfort from the injections (19), symptomatic local weakness (one), and local infection (one) . Preliminary experience indicates that BTX-A is a promising adjunctive treatment for selected patients with spasticity.

J Cell Biol, 1995 Feb, 128(4), 637 - 45
The t-SNAREs syntaxin 1 and SNAP-25 are present on organelles that participate in synaptic vesicle recycling; Walch-Solimena C et al.; Syntaxin 1 and synaptosome-associated protein of 25 kD (SNAP-25) are neuronal plasmalemma proteins that appear to be essential for exocytosis of synaptic vesicles (SVs) . Both proteins form a complex with synaptobrevin, an intrinsic membrane protein of SVs . This binding is thought to be responsible for vesicle docking and apparently precedes membrane fusion . According to the current concept, syntaxin 1 and SNAP-25 are members of larger protein families, collectively designated as target-SNAP receptors (t-SNAREs), whose specific localization to subcellular membranes define where transport vesicles bind and fuse . Here we demonstrate that major pools of syntaxin 1 and SNAP-25 recycle with SVs . Both proteins cofractionate with SVs and clathrin-coated vesicles upon subcellular fractionation . Using recombinant proteins as standards for quantitation, we found that syntaxin 1 and SNAP-25 each comprise approximately 3% of the total protein in highly purified SVs . Thus, both proteins are significant components of SVs although less abundant than synaptobrevin (8.7% of the total protein) . Immunoisolation of vesicles using synaptophysin and syntaxin specific antibodies revealed that most SVs contain syntaxin 1 . The widespread distribution of both syntaxin 1 and SNAP-25 on SVs was further confirmed by immunogold electron microscopy . Botulinum neurotoxin C1, a toxin that blocks exocytosis by proteolyzing syntaxin 1, preferentially cleaves vesicular syntaxin 1 . We conclude that t-SNAREs participate in SV recycling in what may be functionally distinct forms.

Biochem J, 1995 Feb 1, 305 ( Pt 3), 721 - 4
Vesicle-associated membrane protein-2 (synaptobrevin-2) forms a complex with synaptophysin; Washbourne P et al.; Vesicle-associated membrane protein (VAMP) (or synaptobrevin), a type II membrane protein of small synaptic vesicles, is essential for neuroexocytosis because its proteolysis by tetanus and botulinum neurotoxins types B, D, F and G blocks neurotransmitter release . The addition of cross-linking reagents to isolated small synaptic vesicles induces the formation of 30 and 50 kDa complexes containing the isoform 2 of VAMP (VAMP-2) . Whereas the 30 kDa band is a VAMP-2 homodimer, the 50 kDa species results from the cross-linking of VAMP-2 with synaptophysin . This heterodimer also forms in detergent-solubilized vesicles and involves the N-terminal part of VAMP-2 . The implications of the existence of a synaptophysin-VAMP-2 complex in the processes of vesicle docking and fusion with the presynaptic membrane are discussed.

J Otolaryngol, 1995 Feb, 24(1), 64 - 8
Laryngeal image analysis following botulinum toxin injections in spasmodic dysphonia; Wong DL et al.; The injection of botulinum toxin (BT) into laryngeal muscles has proven to be an effective treatment for spasmodic dysphonia (SD), a neurologic disorder characterized by intermittent vocal spasms during speech . BT treatment has reduced abnormal laryngeal muscle hyperactivity in SD patients, when measured by subjective visual rating scales . This paper utilizes images obtained from endoscopic laryngeal video recordings to quantify changes in pre- and post-injection parameters of 17 patients following bilateral BT injections . A previous study, utilizing perceptual, acoustic, and laryngeal aerodynamic measures, showed significant improvements following treatment with BT . In addition, post-injection vocal rest, rather than vocalization, enhanced therapeutic responses . Results of these measures are compared with data from the image analyses and those parameters that proved to be the most useful are highlighted.

Nippon Ganka Gakkai Zasshi, 1995 Feb, 99(2), 232 - 7
{Preoperative prediction of the amount of surgical correction by using Botulinum A type toxin for the treatment of paralytic esotropia}; Iwashige H et al.; The aim of this study was to obtain the correct choice of initial surgical procedure and to predict the amount of surgical correction by using the maximum reduction rate of eye position after Botulinum A-type toxin injection for the treatment of acquired neurogenic paralytic strabismus . The subjects were 30 with sixth nerve palsy and 33 with non-paralytic esotopia with no previous surgery . The results were that if the maximum reduction rate of eye position after the first injection of 1.25 U was less than 68%, the modified Jensen procedure with recession of medial rectus muscle was necessary to achieve 100% recovery . However, if it was more than 100%, then recession-resection of the horizontal rectus muscles alone produced successful ocular alignment . Based on our data of 100 cases of sixth nerve palsy that showed spontaneous recovery, we conclude that surgical treatment should be planned, if the maximum percentage reduction of preoperative eye position after Botulinum A-type toxin treatment is under 68% (reflecting the inability of the medial rectus muscle to contract and the lengthening of the lateral rectus muscle), and if the patient has no spontaneous improvement at all within 12 weeks after onset.

Ann Chir Plast Esthet, 1995 Feb, 40(1), 67 - 76
{Botulinum toxin in the treatment of frontoglabellar and periorbital wrinkles . An initial study}; Ascher B et al.; Glabellar frown lines and crow's feet are wrinkles of facial expression related to an underlying muscular activity, which is particularly strong during facial expression . Classic treatments of these wrinkles only give partially satisfactory are associated with results, and secondary effects, whether they involve skin and muscle lifting, surgical section of muscles, dermal stimulation by thread or injectable fillers, chemical or mechanical abrasion, transient or permanent soft tissue augmentation with various materials . The authors studied the efficacy and safety of intramuscular injections botulinum A Exotoxin in glabellar and crow's feet areas in 19 well-informed and consenting patients . Botulinum toxin injections have been used since 1980 in the treatment of focal dystonia (blepharospasm, oromandibular dystonia, spasmodic torticollis, spasmodic dysphonia and writer's cramp) and safety hemifacial spasm . Their wide use in these indications has highlighted their excellent and efficacy, and the need to repeat injections every 3 to 4 months . The dose required was progressively adjusted around glabellar and orbital areas, while injections of the peri-buccal and forehead areas are still being evaluated . The 19 patients were examined clinically, filmed and photographed every month over a period of 12 to 24 months, and skin prints were performed . Evaluation criteria included the percentage improvement as assessed by the patients themselves, and also evaluation by the investigators of the data of clinical examination, and blind comparison of photographic, videoscopic, and prints . The authors obtained a significant decrease of wrinkles of the areas studied, with a "smoothing" effect during the period of activity of the toxin, which lased an average of 3 to 4 months at the beginning, and 6 to 9 months after several injections . No secondary effects, either general or local, were observed . The product's specificity means that the operator must have a complete mastery of the injection technique and a thorough knowledge of its pharmacology.

Toxicon, 1995 Feb, 33(2), 217 - 27
The median paralysis unit: a more pharmacologically relevant unit of biologic activity for botulinum toxin; Pearce LB et al.; Although the LD50 has been used to quantify the biologically active toxin in clinical preparations of botulinum A toxin (Botox and Dysport), a discrepancy exists between the clinical potency of equivalent international units of different formulations of botulinum A toxin for multiple clinical indications . Our laboratory previously reported that a regional chemodenervation assay in the mouse could be utilized to detect the difference in the potencies of the clinical preparations of toxin {Pearce et al . (1994) Toxic . appl . Pharmac . 128, 69-77} . The purpose of this study was to quantify the regional paralysis produced by botulinum toxin and define a new pharmacologic/biologic unit of activity that more accurately reflects the mechanism of action of botulinum toxin in the clinical setting . Quantal analysis of regional paralysis revealed that the ED50, defined as the median paralysis unit (MPU) for Botox and Dysport, was 0.41 +/- 0.01 and 1.00 +/- 0.02 LD50 units, respectively . Differences in the potencies found in retrospective clinical studies comparing Botox and Dysport were accurately reflected, for the first time, by the dose of toxin expressed in terms of the MPU (median paralysis unit) . The data suggested that the MPU may be a more appropriate measure of the biologic activity in therapeutic formulations of botulinum toxin.

Nurs Times, 1995 Jan 25-31, 91(4), 41 - 3
The effects of botulinum toxin on ocular tissue; Smith H; This report examines the beneficial use of botulinum toxin in the treatment of ophthalmic disorders . It aims to describe the effects and uses of the toxin along with the ophthalmic conditions it is used to treat . The use of botulinum toxin as an effective treatment of ophthalmic disorders is a recent development, with the first treatment occurring in 1983.

J Biol Chem, 1995 Jan 20, 270(3), 1332 - 6
Vesicle-associated membrane protein (VAMP)/synaptobrevin-2 is associated with dense core secretory granules in PC12 neuroendocrine cells; Papini E et al.; The presence and intracellular distribution of vesicle-associated membrane protein-1 (VAMP-1) and VAMP-2 were investigated in the PC12 neuroendocrine cell line using isotype-specific polyclonal antibodies . VAMP-2 was detected in the total membrane fraction, while VAMP-1 was undetectable . Subcellular fractionation demonstrates that a substantial amount of the VAMP-2 (24-36%) is associated with dense core, catecholamine-containing granules (DCGs) . This was confirmed by immunofluorescence microscopy . The L chain of tetanus neurotoxin, known to inhibit granule mediated secretion in permeabilized PC12 cells, as well as botulinum neurotoxins F and G, effectively cleaved DCG-associated VAMP-2 . These data demonstrate that VAMP-2 is present on the secretory granules of PC12 cells.

Biochem Biophys Res Commun, 1995 Jan 17, 206(2), 492 - 6
Possible involvement of protein kinase C and low molecular weight GTP-binding proteins in thrombin-induced histamine secretion in human platelets; Uehara T et al.; Thrombin and 12-O-tetradecanoylphorbol 13-acetate (TPA) caused histamine secretion from human platelets . To clarify the intracellular signalling mechanism of thrombin-induced histamine secretion, the effects of pertussis toxin (PTX) and botulinus toxin (BTX) on thrombin- and TPA-induced histamine secretion were examined in human platelets . The secretion by thrombin was sensitive to BTX, but not PTX . The secretion by TPA was also inhibited by BTX . These results suggest that protein kinase C and low molecular weight G-proteins sensitive to BTX are involved in histamine secretion.

EMBO J, 1995 Jan 16, 14(2), 232 - 9
A role for soluble NSF attachment proteins (SNAPs) in regulated exocytosis in adrenal chromaffin cells; Morgan A et al.; Digitonin-permeabilized chromaffin cells secrete catecholamines by exocytosis in response to micromolar Ca2+ concentrations, but lose the ability to secrete in response to Ca2+ as the cells lose soluble proteins through the plasma membrane pores . Such secretory run-down can be retarded by cytosolic fractions, thus providing an assay for proteins potentially involved in the exocytotic process . We have used this assay to investigate the role of N-ethylmaleimide-sensitive fusion protein (NSF) and soluble NSF attachment proteins (SNAPs) in regulated exocytosis . Recombinant alpha- and gamma-SNAP stimulated Ca(2+)-dependent exocytosis, although recombinant NSF was ineffective, despite the fact that NSF and alpha-SNAP leak from the permeabilized cells with similar time courses . However, around one third of cellular NSF was found to be present in a non-cytosolic form and so it is possible that this is sufficient for exocytosis and that exogenous SNAPs stimulate the exocytotic mechanism by acting on the leakage-insensitive NSF . The stimulatory effect of alpha-SNAP displayed a biphasic dose-response curve and was maximal at 20 micrograms/ml . The effect of alpha-SNAP was Ca(2+)- and MgATP-dependent and was inhibited by N-ethylmaleimide and botulinum A neurotoxin, indicating a bona fide action on the exocytotic mechanism . Furthermore, Ca2+ concentrations which trigger catecholamine secretion acted to prevent the leakage of NSF and alpha-SNAP from permeabilized cells . These findings provide functional evidence for a role of SNAPs in regulated exocytosis in chromaffin cells.

Jpn J Ophthalmol, 1995, 39(4), 424 - 31
Botulinum toxin type A purified neurotoxin complex for the treatment of blepharospasm: a dose-response study measuring eyelid force; Iwashige H et al.; The clinical efficacy of botulinum toxin type A was studied in patients with blepharospasm . Clinical symptoms were evaluated using the Jankovic rating scale . To measure dose response, we used a recently developed device to measure eyelid muscle force . The results showed significant improvement (P = 0.0000) in the Jankovic rating scale scores in all dose groups . The number of patients with marked improvement (6-point decrease or more in the total Jankovic rating scale score) increased with higher dose injections . After injections of 0.50, 1.25, or 2.50 U/site, 6 sites/eye, the eyelid muscle force decreased by 33.2 +/- 28.1%, 41.7 +/- 25.1%, or 69.6 +/- 5.0%, respectively . The decrease of eyelid muscle force showed a significant dose response (P = 0.0254) . The mean duration of effect was 12.9 weeks in patients after dose injections of 1.25 U/site, which was significantly longer (P = 0.0205) than the 9.6 weeks in patients after dose injections of 0.50 U/site . No severe adverse effects were observed . We concluded that injections of botulinum toxin type A at an initial dose of 1.25 U/site are a safe and effective treatment for blepharospasm.

J Fr Ophtalmol, 1995, 18(12), 751 - 7
{Clinical and psychological factors influencing the efficacy of botulinum toxin in the treatment of hemifacial spasm and blepharospasm}; Burbaud P et al.; We analyzed the influence of clinical and psychological factors on the long-term efficacy of botulinum toxin A (BTX) injections in 45 patients with blepharospasm and 66 patients with hemifacial spasm . Injections efficacy (respectively 94.3% and 95.7%) and duration of relief (respectively 14.8 and 18.7 weeks) remained stable over seven successive injections . Clinical improvement was not influenced by patients' sex, age, or disease duration but by psychological background (p < 0.001) . Patients who failed to respond after repetitive injections had lower inter-injections intervals (p < 0.05) . This data shows: (1) the importance of psychological contexte in subjective evaluation of treatment efficacy with BTX, (2) emphasizes the necessity of avoiding close injections.

Arch Med Res, 1995 Winter, 26(4), 405 - 8
Botulinum toxin-A for the treatment of hemifacial spasm; Cuevas C et al.; Management of hemifacial spasm can actually be done medically, surgically and with Botulinum-A Toxin . The Botulinum-A Toxin treatment locally injected into the involved facial muscles offers a useful alternative to medical and surgical therapy . The objective of this study was to evaluate the efficacy of Botulinum-A Toxin for the treatment of hemifacial spasm in those subjects for whom the presently available medical therapy is inadequate . A total of 28 individuals were enrolled in the clinical study . Patients were evaluated using the Fahn's blepharospasm rating and disability scales . Efficacy was assessed by evaluating changes from the baseline in eyelid spasm intensity, brow spasm intensity, eyelid force and facial spasm intensity . All 28 subjects with hemifacial spasm showed clinical improvement in relation to this baseline, which was statistically significant . The mean decrease from baseline at their follow-up examination was statistically significant for all subjects and for all measurements: eyelid spasm changed from 2.3 to 0.3 (p = 0.0001); brow spasm from 1.9 to 0.1 (p = 0.0001); facial spasm from 2.3 to 0.1 (p = 0.0001) and eyelid force from 0.9 to -0.1 (p = 0.0020) . We concluded that Botulinum-A Toxin provides a significant therapeutic benefit to patients with hemifacial spasm, without the risk of disabling side effects.

Rev Neurol, 1995 Jan-Feb, 23(119), 129 - 33
{Syndromes of continuous muscular activity: report of a central case (stiff-man) and a peripheral case (neuromyotonia) associated with neuroborreliosis}; Requena I et al.; We describe two cases of continuous muscular activity: one which is central (the stiff-man syndrome), and another which is peripheral (neuromiotony), the latter in a patient suffering from diabetic neuropathy and with positive Borrellia burgdorferi serology in the bloodstream, as well as CSF . Both cases reacted favourably to medical treatment . In the first case botulinic toxin was used as a simultaneous treatment for focal pseudodystonia in one foot . Response was good.

Eye, 1995, 9 ( Pt 5), 558 - 63
Management of strabismus due to orbital myositis; Bessant DA et al.; We report on 5 consecutive patients seen at the botulinum toxin clinic at Moorfields Eye Hospital with an ocular motility disorder secondary to orbital myositis . CT scans demonstrated involvement of one or both of the medial recti in the inflammatory process in all 5 patients . In addition 1 patient had involvement of both the lateral recti and the right superior rectus . Two patients had been treated with oral steroids, 3 with non-steroidal anti-inflammatory agents, and 1 with orbital radiotherapy . Prior to toxin injection 3 patients had an esotropia (ranging from 4 delta to 30 delta) and two an exotropia (52 delta and 85 delta) . A vertical imbalance was present in 3, and all 5 patients had symptomatic diplopia . A total of six injections were given to 5 patients, 2 of whom later went on to have surgery . Toxin injection reduced the angle of the deviation to less than 10 delta in 4 patients, all of whom are now asymptomatic . The fifth patient has persistent diplopia despite two operations to correct a large exotropia . We discuss the role of botulinum toxin and surgery in the management of strabismus due to orbital myositis.

J Physiol Paris, 1995, 89(2), 83 - 94
Role of myosin in neurotransmitter release: functional studies at synapses formed in culture; Mochida S; To determine the functional role of presynaptic proteins in the neurotransmitter release, I have employed cholinergic synapses formed between superior cervical ganglion neurons in culture . These synapses expressed proteins characteristic of mature synapses: immunofluorescence staining showed the presence of synaptophysin, synaptotagmin, VAMP/synaptobrevin-2, syntaxin and neurexin . The function of these proteins seems to be similar to that of mature synapses because botulinum neurotoxins A, E and C1 inhibited neurotransmitter release evoked by presynaptic action potentials . With this preparation, I have obtained evidence supporting roles for myosin II and myosin light chain kinase in neurotransmitter secretion . Acetylcholine release was inhibited by introduction of antibody against myosin II or inhibitors of myosin light chain kinase . This evidence suggests a model in which myosin light chain kinase phosphorylates myosin, and the resultant change in actin-myosin interactions is involved in some steps of transmitter release.

Neuron, 1995 Jan, 14(1), 133 - 41
Nerve sprouting in muscle is induced and guided by processes extended by Schwann cells; Son YJ et al.; Partial denervation or paralysis with botulinum toxin, manipulations that induce sprouting of nerve terminals in muscle, also induced terminal Schwann cells to extend processes . These processes were associated with every nerve sprout and in some cases were longer than the sprouts that appeared to be growing along them . Following partial denervation, more than 70% of the nerve sprouts that grew to innervate nearby denervated endplates were associated with Schwann cell processes that had extended from the denervated endplates, i.e., in the direction opposite to nerve growth . Implantation of Schwann cells into an innervated muscle induced sprouting upon contact of an axon or nerve terminal by Schwann cell processes . These observations show that Schwann cells induce and guide axonal sprouting in muscle.

J Neurosci, 1995 Jan, 15(1 Pt 2), 520 - 8
Calcitonin gene-related peptide: possible role in formation and maintenance of neuromuscular junctions; Sala C et al.; The expression and content of calcitonin gene-related peptide (CGRP) and secretogranin II (SgII) in adult rat motor neurons were examined by in situ hybridization, Northern blot analysis, and immunocytochemistry . Normal motor nerve terminals did not contain detectable CGRP or SgII . Ten to 15 days after a peripheral nerve crush about 80% of the motor nerve terminals reinnervating the soleus (SOL) muscle contained detectable CGRP but no SgII . Thereafter, the percentage of CGRP-positive terminals declined towards zero . In the spinal cord, CGRP expression was higher than normal 1 d after a sciatic nerve crush and increased during the next few days . No increase in SgII expression was observed . Nerve blocks by tetrodotoxin (TTX) and botulinum toxin (BoTX) increased CGRP content and expression in motor neurons but had no effect on SgII . After 10 d of BoTX treatment and 33 d of TTX treatment (the longest time points studied), more than 90% of the motor nerve terminals stained for CGRP . The density of large dense core vesicles (LDCVs) was also higher than normal in such terminals . Some increase in CGRP content and expression occurred in the nontreated side . In a group of rats, the peroneal nerve was stimulated electrically with brief, intermittent pulse trains at 100 Hz . The stimulation was applied below a TTX block that had started 7 or 19 d earlier . One minute of such stimulation was sufficient to remove CGRP from most of the terminals.(ABSTRACT TRUNCATED AT 250 WORDS)

Hokkaido Igaku Zasshi, 1995 Jan, 70(1), 19 - 28
{Structure and function of botulinum toxin}; Fujii N; Botulinum toxins (types A to G) inhibit the release of acetylcholine at the neuromuscular junction . These toxins are produced as progenitor toxins of large molecular sizes of 12S (M toxin), 16S (L toxin) and 19S (LL toxin) in culture supernatants . Three different molecular forms have been demonstrated in botulinum type A toxin . L and M toxins are recognized in botulinum type C and D toxins . Type E toxin is exclusively composed of M toxin . In an alkaline condition, M and L toxins dissociate into neurotoxin and nontoxic components . Nontoxic components consist of nontoxic-nonhemagultinin component (nontoxic-nonHA) and hemagultinin (HA) . M toxin is made up by association of neurotoxin with nontoxic-nonHA, and L toxin is formed by conjugation of M toxin with HA . HA also consists of several subcomponents . These genes with related functions (progenitor toxin) are closely grouped as operon on the chromosome . Nontoxic-nonHA gene is located only 17 bp (type C) or 27 bp (type E) upstream of the neurotoxin gene . Both genes may be transcribed (right-ward transcription) by a polycistronic mRNA species initiated from a promoter located in the 5'-untranslated region of the nontoxic-nonHA gene . The construction of HA subcomponent genes (HA-33, HA-17, HA-25 and HA-53) also appears operon structure . The gene cluster related HA is located 262 bp upstream of nontoxic-nonHA gene of type C and transcribed (left-ward transcription) by the same mRNA from the 5'-noncoding region of HA-33 gene . Botulinum neurotoxin undergoes cleavage to form a dichain molecule linked through a disulphide bond . The heavy chain correlates with the binding of toxin to peripheral synapses, and the light chain is associated with the intracellular activity of blocking of acetylcholine release . Fifty amino acids in C-terminal region of type C toxin is essential for the binding activity of toxin to the target cells . However, the binding efficiency of type C toxin is not antagonized by the other type of botulinum toxins because of low homology of this binding domain of type C toxin to other types . Furthermore, five highly homologous regions are found in light chain among seven neurotoxins . One of these homologous regions, sequence HEL-H--, shows strong similarity with the active site of zinc-proteases . The inhibition of acetylcholine release is associated with this protease activity which selectively cleaves the synaptic vesicle membrane proteins . These target membrane proteins are key components of the synaptic vesicle docking and fusion.(ABSTRACT TRUNCATED AT 400 WORDS)

Gig Sanit, 1995 Jan-Feb, (1), 9 - 12
{Stability of botulin toxins in solutions and beverages}; Kazdobina IS; Stability of botulinic toxins of types A, B, C, E and F in drinking water, various solutions, and beverages was studied . The stability of all toxins was the highest in bear, mineral and fruit waters, dessert wine and 40 degrees alcohol, where toxins preserved 50% of initial toxicity during 5-70 days . C-type toxin was the most stable in drinking water . Biological assay and neutralization test on white mice can be used for detection of botulinic toxins.

Eur Neurol, 1995, 35(1), 43 - 5
Treatment of hemifacial spasm with botulinum toxin . Value of preinjection electromyography abnormalities for predicting postinjection lower facial paresis; Angibaud G et al.; Thirty-two patients with hemifacial spasm were treated with 61 botulinum toxin (BT) injections . Some patients had post-BT lower facial paresis (LFP+group) while others did not suffer this side effect (LFP-group) . Abnormal electromyography (EMG) recordings were more frequent in the LFP+ group (6/11) than in the LFP- group (3/21; p < 0.05) . This result suggests that an EMG performed before any BT injection could detect the patients with a higher risk for this side effect . The first dose of BT might be lowered in such cases.

Va Med Q, 1995 Summer, 122(3), 184 - 5
Achalasia . A new modality for treatment; Mitchell RE Jr et al.; This lesion of the esophagus, first described in 1682 by Thomas Willis, been subject to many forms of therapy . We feel botulinum toxin injection to be an acceptable alternative treatment modality for select patients with primary esophageal achalasia . Traditional methods of treating achalasia consist of medical therapy for short-term relief, balloon dilation and myotomy . Botulinum toxin injection is an alternative method of treatment, suggested by Pasricha et al and used successfully in our patient, which does not seem to cause the significant complications of perforation or gastroesophageal reflux and which may be more attractive to patients less able to undergo dilation or myotomy . This method of injecting botulinum toxin directly into the LES appears to be a relatively safe modality of treatment . Reports suggest symptoms of achalasia may recur (in up to a year's time) and repeated injections may be needed . Even so, this would seem to be acceptable in the overall management of achalasia . We agree that long-term follow-up of these patients is indicated . Data to date, plus our personal experiences, have been encouraging . We feel this represents an option for non-surgical patients and may even be considered prior to the endoscopic balloon surgery approach . It is certainly more cost effective . We are currently evaluating a second patient for botulinum toxin therapy.

Biochemistry, 1994 Dec 27, 33(51), 15365 - 74
Differences in the protease activities of tetanus and botulinum B toxins revealed by the cleavage of vesicle-associated membrane protein and various sized fragments; Foran P et al.; Botulinum neurotoxin serotype B (BoNT/B) and tetanus toxin (TeTx) block neuroexocytosis through selective endoproteolysis of vesicle-associated membrane protein (VAMP) . The enzymological properties of both toxins were compared for the first time in their cleavage of VAMP and various sized fragments using a sensitive chromatographic assay . The optimal substrate sizes for the zinc-dependent protease activities of the light chains of TeTx and BoNT/B were established using synthetic peptides corresponding to the hydrophilic core of VAMP (30-62 amino acids in length) . TeTx was found to selectively cleave the largest peptide at a single site, Gln76-Phe77 . It exhibited the most demanding specificity, requiring the entire hydrophilic domain (a 62-mer) for notable hydrolysis, whereas BoNT/B efficiently cleaved the much smaller 40-mer . Thus, an unusually long N-terminal sequence of 44 amino acids upstream of the scissile bond is required for the selective hydrolysis of VAMP by TeTx . Using the largest peptide, BoNT/B and TeTx exhibited approximately 50% and 35%, respectively, of the activities shown toward intact VAMP, detergent solubilized from synaptic vesicles . Given the large size of the smallest substrates, it is possible that these neurotoxins recognize and require a three-dimensional structure . Although both toxins were inactivated by divalent metal chelators, neither was antagonized by phosphoramidon or ASQFETS (a substrate-related peptide that spans the cleavage site), and TeTx was only feebly inhibited by captopril; also, they were distinguishable in their relative activities at different pHs, temperatures, and ionic strengths . These collective findings are important in the design of effective inhibitors for both toxins, as well as in raising the possibility that TeTx and BoNT/B interact somewhat differently with VAMP.

Otolaryngol Head Neck Surg, 1994 Dec, 111(6), 787 - 94
Technique for injection of botulinum toxin through the flexible nasolaryngoscope; Rhew K et al.; A new endoscopic method of injecting botulinum toxin into the thyroarytenoid muscles for treatment of adductor spasmodic dysphonia was evaluated . Twelve patients with adductor spasmodic dysphonia were given injections in the thyroarytenoid muscle under video visualization with a flexible catheter needle that was passed through the working channel of a flexible nasolaryngoscope . Six patients received unilateral injections, and six received bilateral injections . Preinjection and postinjection speech samples were compared by use of spectrographic analysis . Significant decreases in voice breaks and sentence duration were found after treatment with both unilateral and bilateral injections . Patient interviews and diaries documented the reported degree and duration of symptom reduction . All 12 patients reported that the injections were of significant benefit and that the endoscopic procedure was tolerable . We concluded that this is a safe and effective technique for injecting botulinium toxin into laryngeal muscles for treatment of spasmodic dysphonia.

Neurology, 1994 Dec, 44(12), 2262 - 6
Laryngeal botulinum toxin injections for disabling stuttering in adults; Brin MF et al.; Stuttering is an action-induced speech disorder with involuntary, audible, or silent repetitions or prolongations in the utterance of short speech elements (sounds, syllables) and words . Symptomatic treatment programs frequently have initial success; persistent benefit is variable and many patients remain disabled . Stuttering has many characteristics similar to spasmodic dysphonia (laryngeal dystonia), often including the presence of adductor laryngeal spasms that obstruct airflow (glottal block) . We hypothesized that relief of the spasmodic dysphonic glottal blocks in stutterers would modify the stuttering phenomenon and increase fluency . We therefore studied the effects of bilateral vocal fold injections of botulinum toxin type A (BTX) on dysfluency and speech characteristics in stuttering . We treated 14 adult patients (12 men, 2 women) with persistent stuttering and glottal block who previously failed standard speech therapy with 1.25 U BTX into each thyroarytenoid (vocalis) muscle . Fluency evaluations included the Stuttering Severity Instrument, the Perceptions of Stuttering Inventory, and a global rating scale (percent of normal function) . Patients were evaluated at baseline and at 2-, 6-, and 12-week follow-up visits . Improvement in fluency documented by each rating instrument occurred at 2 and 6 weeks, with functional relapse by 12 weeks in most patients . We conclude that therapeutic laryngeal injections of botulinum toxin are useful in the management of stuttering with glottal block and result in a moderate improvement in fluency . When an adult patient with developmental stuttering with glottal blocks has failed speech interventional therapy and presents for treatment, a trial of BTX can be considered early.(ABSTRACT TRUNCATED AT 250 WORDS)

Oncogene, 1994 Dec, 9(12), 3519 - 26
The regulation of endothelial cell motility by p21 ras; Fox PL et al.; Directed endothelial cell (EC) movement is required for the development and repair of blood vessels and plays a critical role in angiogenic processes obligatory for large tumor formation . We now report that ras proteins have a critical role in regulation of movement of normal mammalian cells . Bovine aortic EC microinjected with oncogenic Ha-ras enter further into an artificial wound than uninjected cells . Treatment with oncogenic Ha-ras also converts the cell paths from nearly linear in control cells to apparent 'random-walk' trajectories in treated cells, suggesting that oncogenic ras alters the normal control processes regulating cell motility . Botulinum toxin C blocks ras-stimulated motility indicating that a member of the p21 rho family is a downstream participant in the motile pathway . In related experiments we have observed that microinjection of the neutralizing, ras-specific, Y13-259 monoclonal antibody completely blocks both basal and basic fibroblast growth factor-stimulated movement of aortic EC . Y13-259 blocks the initiation of EC movement, as well as the continued progress of cells already in motion, suggesting that ras activity is continuously required throughout the motile process . Together these data indicate that ras is an integral component of the signaling pathway regulating cell movement.

Muscle Nerve, 1994 Dec, 17(12), 1385 - 92
Serial neurophysiological studies of intramuscular botulinum-A toxin in humans; Hamjian JA et al.; To characterize the time course of intramuscular botulinum toxin-induced paresis, we serially performed electrophysiological measurements and recorded the sonographic size of an extensor digitorum brevis (EDB) muscle in 10 human subjects before and after injecting the EDB with 10 units of botulinum-A toxin . All EDB CMAPs decreased within 48 h, with peak decline at day 21 (8.3 +/- 3.1 mV to 3.0 +/- 0.9 mV) . Decline of mean rectified voltage during maximal voluntary contraction of the EDB paralleled the change in CMAP amplitude . Average decrements to 2-Hz repetitive stimulation never exceeded 6% (day 42) and exercise failed to facilitate significantly CMAP amplitude . Atrophy peaked at day 42 . The F-wave to M-wave ratio increased at day 2; silent periods did not change . Our findings confirm a primary peripheral action of the toxin, but a superimposed, transient central effect of the drug cannot be excluded . Intramuscular injections into EDB provide a useful model for studying chemodenervation effects.

J Neuroophthalmol, 1994 Dec, 14(4), 199 - 204
Chemomyectomy of the orbicularis oculi muscles for the treatment of localized hemifacial spasm; Wirtschafter JD; OBJECTIVE: To report our experience with doxorubicin chemomyectomy as an alternative to other treatments for hemifacial spasm (HFS) . DESIGN: A prospective, open study Phase I clinical trial of chemomyectomy . SETTING: A hospital-based, referral neuro-ophthalmology and oculoplastic service . PATIENTS AND METHODS: Repeated (1-6, median: 4) local injections of doxorubicin were given in the eyelids of 8 patients (5 men, 3 women, average age: 71) . MAIN OUTCOME MEASURES: Eyelid strength, self-reported spasm, and duration of improvement without seeking additional or alternative treatments . RESULTS: Chemomyectomy resulted in permanent (> or = 2.5 years) orbicularis oculi weakness and relief from spasms in the treated areas in 5 patients, although 2 patients requested occasional supplementary botulinum toxin (BT) injections in the facial muscles over the cheek . One patient had a successful result for 3 years, after which spasm recurred . One patient maintains a successful result in the eyelid but had a failed microvascular decompression in the lower face . One incompletely treated patient required microvascular decompression following spread of spasms to the lower branches of the facial nerve and increased severity of the HFS . One patient required eyelid surgery because of concurrent spastic entropion . One patient treated with a higher concentration than currently used required closure of a skin ulcer . CONCLUSIONS: Doxorubicin chemomyectomy is an effective alternative to conventional therapy for properly selected patients affected by HFS, particularly older patients with relatively localized eyelid muscle spasms . The modified technique of doxorubicin chemomyectomy has developed to the point where its safety is demonstrated and its risks are known.

Scand J Rehabil Med, 1994 Dec, 26(4), 191 - 5
Efficacy of botulinum toxin for cervical dystonia . A comparison of methods for evaluation; Odergren T et al.; Twenty patients with cervical dystonia were treated during one year with repeated intramuscular injections of botulinum toxin . The outcome was evaluated comparing subjective global rating with relative changes in degree of pain on the Visual analogue scale (VAS), degree of dysfunction due to dystonia, and quality of life according to the Nottingham health profile (NHP) . Objective measurement of dystonic position and movement ability was performed using a goniometer, semiquantitatively noted as scores according to Fahn and Tsui . Before treatment, the degree of impaired life quality on the NHP did not correlate with the Tsui score of dystonic posture, but significantly with the Fahn score (p < 0.01) which also includes data on pain . Significant improvement after treatment was seen for all parameters (p < 0.05) . Global subjective rating correlated significantly with improved posture according to the Tsui score (p < 0.05), but not with reduced pain or degree of dysfunction . The results suggest that the efficacy of botulinum toxin in cervical dystonia is best evaluated using a combination of the VAS for pain and the Tsui score for dystonic posture and movement ability.

Z Gastroenterol, 1994 Dec, 32(12), 694 - 701
{Diagnostic and therapeutic possibilities in suspected Oddi's sphincter dysfunction}; Wehrmann T et al.; Endoscopic manometry and quantitative cholescintigraphy are the diagnostic cornerstones for the detection of suspected sphincter of Oddi dysfunction . In patients with recurrent biliary pain after cholecystectomy, endoscopic manometry proves an elevated sphincter of Oddi baseline pressure as the most common finding . The probability for the detection of an elevated baseline pressure in these patients is significantly correlated with the presence of certain clinical features (i.e . biliary pain and/or cholestasis and/or dilated bile duct and/or delayed drainage of contrast material after ERCP) . Therefore, these features enable a clinical classification of patients with suspected sphincter of Oddi dysfunction . Isolated baseline pressure elevations in the pancreatic portion of the sphincter of Oddi were reported in patients with recurrent, idiopathic, acute pancreatitis . In patients with biliary sphincter dysfunction, therapeutic relief can be expected from pharmacological therapy, but controlled studies are lacking . However, the clinical value of endoscopic sphincterotomy could be established in this field . Despite endoscopic manometry is not a prerequisite for the performance fo endoscopic sphincterotomy in every case of suspected sphincter of Oddi dysfunction, in most patients endoscopic manometry allows the only definitive diagnosis of sphincter dysfunction . Further on, the clinical value of semi-invasive methods as alternative treatment strategies (i.e . botulinum-toxin, transcutaneous electric nerve stimulation, balloon dilation) for sphincter of Oddi dysfunction has to be evaluated in the future.

J Voice, 1994 Dec, 8(4), 347 - 51
Botulinum toxin in the treatment of recalcitrant mutational dysphonia; Woodson GE et al.; Mutational falsetto is the failure of the normal drop in vocal pitch at puberty . Voice therapy almost always achieves an appropriate pitch; however, in cases of failure, surgical treatment has also been recommended . We report a case of a 47-year-old man with an above-average fundamental frequency and a thin voice quality in the absence of any signs of androgen insufficiency . Laryngeal examination revealed atrophy of the vocalis muscle . Voice therapy was unsuccessful in achieving a stable voice . Injection of 15 units of botulinum toxin into each cricothyroid muscle initially resulted in aphonia, but the voice returned by 1 week . Average fundamental frequency was 84 Hz at 1 week, 104 Hz at 1 month, and 100 Hz at 1 year . We hypothesize that mutational dysphonia is an habitual dysfunction of the voice with inappropriate activation of the cricothyroid muscle and disuse of laryngeal adductor muscles . Temporary deactivation of the cricothyroid muscle enforces adoption of a more appropriate vocal mechanism . Botulinum toxin as an adjunct to voice therapy should be considered before surgical alteration of the glottis in patients with recalcitrant mutational falsetto.

Brain, 1994 Dec, 117 ( Pt 6), 1457 - 74
Abnormal eye movements in blepharospasm and involuntary levator palpebrae inhibition . Clinical and pathophysiological considerations; Aramideh M et al.; We report on four patients with involuntary eyelid closure and eye movement disorders . Three were healthy until the onset of their illness and one had a mild generalized choreoathetosis and dystonia due to kernicterus . Electromyographic recording revealed solely blepharospasm in two patients and blepharospasm in combination with involuntary levator palpebrae inhibition in the other two . The eye movement abnormalities were clinically characterized by inability to fix gaze and short or prolonged episodes of uncontrollable eye deviations accompanied, in two patients, by diplopia in horizontal or vertical directions . These episodes occurred independently of a disorder of eyelid movement . Eye movement recordings with a double magnetic induction technique showed saccadic intrusions in horizontal directions . They consisted of highly frequent square wave jerks in three and sporadic macro-square wave jerks in two patients . There were also episodes of extraocular muscle dystonia, commonly known as oculogyric crises, resulting in involuntary upward eye deviation in all patients and lateral deviation in three patients . In one patient, nasal-ward deviations were sometimes restricted to one eye . We conclude that these abnormal eye movements do not necessarily point to a symptomatic form of dystonia and that they may limit the beneficial effect of botulinum toxin or surgical intervention in the therapeutic management of involuntary eyelid closure . We suggest that either basal ganglia, especially substantia nigra pars reticularis and the brainstem structures, especially the paramedian pontine reticular formation, or both, may be involved in the pathogenesis of these abnormal movements.

J Neurol Neurosurg Psychiatry, 1994 Dec, 57(12), 1535 - 7
Optimisation of botulinum treatment for cervical and axial dystonias: experience with a Japanese type A toxin; Mezaki T et al.; Twenty two patients with cervical and axial dystonias were treated with Japanese type A botulinum toxin . Injections were given repeatedly at intervals of 28-30 days to carefully chosen muscles with increased activities, with a maximum dose per session of 300 units . The maximum improvements in subjective and objective ratings were obtained only after repeated injections . No anti-toxin antibodies were detected; nor did any muscle fail to respond to the toxin . During the treatment, previously "silent" muscles were activated to reproduce the original abnormal posture, as if driven by a central motor programme . This resistance to treatment was overcome by injecting the toxin into newly activated muscles . Repeated injections are thus required to override central mechanisms in dystonias or to maximise drug delivery to large muscles . Antibody development may be controlled by the use of a less immunogenic toxin.

Drugs, 1994 Dec, 48(6), 888 - 93
Botulinum toxin in clinical practice; Hughes AJ; Over recent years botulinum toxin type A has emerged as a safe and effective treatment for a number of previously refractory conditions associated with excessive muscle activity . The list of indications is expanding, but at present it is generally considered to be the treatment of choice for focal dystonias such as blepharospasm, torticollis, laryngeal dystonia, and oromandibular dystonia, as well as hemifacial spasm, strabismus, and some forms of limb spasticity . Carefully targeted intramuscular injections of a small amount of the toxin block the release of acetylcholine at the neuromuscular junction, producing a chemical denervation, with the aim of reducing excessive muscle activity without producing significant functional weakness . In some situations electrophysiological assessment and localisation of the muscles for injection is necessary . Treatment is symptomatic, with effects lasting 3 to 4 months and most patients requiring up to 4 injections per year to maintain the beneficial effect . Appropriate use of the toxin requires both an understanding of the physiological action of the potential muscles involved in each situation, together with a knowledge of the likely dose necessary to reduce muscle activity to the required level . Botulinum toxin represents a major advance in the management of these conditions, many of which responded poorly to previously available forms of therapy.

Electroencephalogr Clin Neurophysiol, 1994 Dec, 93(6), 434 - 9
Quantitative EMG in botulinum toxin treatment of cervical dystonia . A double-blind, placebo-controlled study; Ostergaard L et al.; We used turns-amplitude analysis of the EMG as a guidance for botulinum toxin (BT) treatment in 19 patients with cervical dystonia . At the first examination, muscles showing abnormal activity (> 100 turns/sec at rest) were given BT 75 units (10 patients) or placebo (9 patients) . At subsequent examinations, about 6, 12 and 18 weeks after the start, BT 75 units were given to all hyperactive muscles . Six weeks after the first BT treatment the sternocleidomastoid muscle contralateral to the involuntary head rotation and the ipsilateral and contralateral posterior neck muscles (PNM) showed a reduction of involuntary activity, as indicated by reduced turns/sec and mean amplitude at rest . Similar changes were seen when comparing BT treatment with placebo . The reduction was greater in the contralateral sternocleidomastoid muscle than in PNM, suggesting that PNM need higher doses of BT . At maximal voluntary contraction, BT treated muscles showed unchanged turns/sec (5/6 tests), decreased mean amplitude and increased ratio (turns/amplitude) . This may reflect a functional random loss of muscle fibres, combined with inability to activate all motor units . A high (89%) clinical success rate with BT therapy was obtained, and it is concluded that quantitative EMG is a useful tool for the precise identification of hyperactive muscles, for optimal placing of the injection cannula and for unbiased monitoring of the treatment effect.

Biochem Biophys Res Commun, 1994 Nov 30, 205(1), 751 - 7
The light chain of botulinum neurotoxin forms channels in a lipid membrane; Kamata Y et al.; The ability of botulinum neurotoxin and its isolated subunits, the heavy and light chains, to bind to a lipid membrane and to form channels in the membrane was examined . At pH 4.0, the neurotoxin caused aggregation of calcein-containing liposomes, providing evidence of binding of the neurotoxin to the surface of the outer lipid membrane . Aggregation was followed by the release of calcein, as a result of the formation of channels . The heavy chain evoked the same responses as those of the neurotoxin . The light chain did not cause aggregation of the liposomes but did evoke the release of calcein . The channel-forming ability of the light chain appeared to be higher than that of the neurotoxin or the heavy chain . This novel property of the light chain may help us to understand the mechanism of action of botulinum neurotoxin.

J Biol Chem, 1994 Nov 4, 269(44), 27427 - 32
SNAP-25, a t-SNARE which binds to both syntaxin and synaptobrevin via domains that may form coiled coils; Chapman ER et al.; The membrane proteins SNAP-25, syntaxin, and synaptobrevin (vesicle-associated membrane protein) have recently been implicated as central elements of an exocytotic membrane fusion complex in neurons . Here we report that SNAP-25 binds directly to both syntaxin and synaptobrevin . The SNAP-25-binding domain of syntaxin lies between residues 199 and 243, within the region previously shown to mediate synaptobrevin binding (Calakos, N., Bennett, M . K., Peterson, K . E., and Scheller, R . H . (1994) Science 263, 1146-1149) . The syntaxin-binding domain of SNAP-25 encompasses most of the amino-terminal half of SNAP-25, including its putative palmitoylation sites . Truncation of the carboxyl-terminal 9 residues of SNAP-25, which yields a fragment corresponding to that generated by botulinum neurotoxin A, diminishes the interaction of SNAP-25 with synaptobrevin, but not with syntaxin . Sequence analysis revealed that the regions that mediate the interaction between SNAP-25 and syntaxin contain heptad repeats characteristic of certain classes of alpha-helices . Similar repeats are also present at the carboxyl terminus of SNAP-25 and in synaptobrevin . These domains have a moderate to high probability of forming coiled coils . We conclude that SNAP-25 can interact with both syntaxin and synaptobrevin and that binding may be mediated by alpha-helical domains that form intermolecular coiled-coil structures.

Cell, 1994 Nov 4, 79(3), 507 - 13
The small GTP-binding protein Rho regulates a phosphatidylinositol 4-phosphate 5-kinase in mammalian cells; Chong LD et al.; Integrin-mediated adhesion is known to stimulate production of phosphatidylinositol 4,5-bisphosphate (4,5-PIP2) and increase 4,5-PIP2 hydrolysis in response to platelet-derived growth factor (PDGF) . We now show that treatment of cells with lovastatin, which inhibits modification of small GTP-binding proteins, reduced PIP2 levels and decreased calcium mobilization in response to PDGF and thrombin . In cell lysates, GTP gamma S stimulated PIP 5-kinase activity, and this effect was blocked by botulinum C3 exoenzyme, suggesting that Rho was responsible . GTP-bound recombinant Rho stimulated PIP 5-kinase activity, whereas GDP-Rho was much less potent and GTP-bound Rac was ineffective . Microinjected botulinum C3 exoenzyme caused diminished calcium mobilization in response to PDGF or thrombin . Conversely, microinjection of activated Rho reversed the decrease in calcium mobilization normally seen in nonadherent cells . These data demonstrate that Rho regulates 4,5-PIP2 synthesis and, indirectly, 4,5-PIP2 hydrolysis . They also raise the possibility that PIP2 synthesis could mediate the effects of Rho on the actin cytoskeleton.

Isr J Med Sci, 1994 Nov, 30(11), 816 - 9
Interventional neurology: botulinum toxin as a potent symptomatic treatment in neurology; Giladi N et al.; Local injections of botulinum toxin is a well-accepted treatment for focal dystonias, hemifacial spasms and strabismus . Its use by skilled neurologists has been reported to be safe and effective . We report our experience with botulinum toxin injections in 108 patients with various central nervous system disorders . Botox was effective in upper face dystonia (86% improvement), spastic dysphonia (92% improvement), platysma muscle spasms and spasmodic torticollis (range of movement 61%, pain and tension 90%) . It was also very effective in a few patients with apraxia of eyelid opening, parkinsonian jaw tremor, teeth clenching, palatal myoclonus and adductor leg spasticity . No serious side effects were recorded . Botulinum toxin is a useful symptomatic treatment for many neurological disorders, and one of the leading mode of treatments in the new subspecialty in neurology called "Interventional neurology."

J Neurol Neurosurg Psychiatry, 1994 Nov, 57(11), 1321 - 4
Botulinum toxin treatment for lower limb extensor spasticity in chronic hemiparetic patients; Hesse S et al.; Twelve chronic hemiparetic outpatients with pronounced lower limb extensor spasticity were injected with 400 units of botulinum toxin A, EMG guided into the soleus, tibialis posterior, and both heads of the gastrocnemius muscles . Botulinum toxin A caused a definite reduction of plantar flexor spasticity, in 10 patients two weeks after the injection, as assessed by the Ashworth scale . Four of the patients were able to achieve active dorsiflexion of their affected ankle . Gait analysis including the measurement of vertical ground reaction forces showed a statistically significant (p < 0.01) improvement in velocity, stride length, stance symmetry, and the length of the force point of action under the affected foot . Qualitative improvements on the force diagrams indicated a better loading, advancement of the body, and push off of the affected limb in seven patients . Eight weeks after the injection the effects waned.

J Biochem (Tokyo), 1994 Nov, 116(5), 1134 - 8
ADP-ribosylarginine glycohydrolase catalyzing the release of ADP-ribose from the cholera toxin-modified alpha-subunits of GTP-binding proteins; Maehama T et al.; A rat glycohydrolase which catalyzes the hydrolysis of ADP-ribosylarginine was expressed in Escherichia coli and purified to homogeneity for characterization of its enzymatic properties . The purified glycohydrolase catalyzed the hydrolysis of N-glycoside linked ADP-ribosylarginine on the alpha-subunits of stimulatory GTP-binding proteins (Gs) and cholera toxin A1-subunit that had been modified by cholera toxin and NAD . Nonmuscle actin of which an arginine residue was ADP-ribosylated by botulinum C2 toxin also served as a substrate of the glycohydrolase . On the other hand, the glycohydrolase did not hydrolyze ADP-ribosylated cysteine on the alpha-subunits of pertussis toxin-substrate GTP-binding proteins, ADP-ribosylated diphthamide on elongation factor 2, or ADP-ribosylated asparagine on rho GTP-binding proteins . The rate of the reaction catalyzed by the glycohydrolase was affected by nucleotide-binding form of the ADP-ribosylated substrate proteins; the GDP-bound form of the modified Gs-alpha was more rapidly hydrolyzed than the guanosine 5'-(3-O-thio)triphosphate-bound form . Interestingly, the glycohydrolase activity was markedly inhibited by mM order concentration of ATP in addition to ADP-ribose, the product of the enzyme reaction, though ADP had no inhibitory effect on the activity . Moreover, alpha NAD, but not beta NAD, inhibited the enzyme activity, suggesting that the glycohydrolase reaction was stereospecific for the alpha-anomer.

Mov Disord, 1994 Nov, 9(6), 610 - 5
"Apraxia of lid opening," a focal eyelid dystonia: clinical study of 32 patients; Krack P et al.; We have seen 32 patients with "apraxia of lid opening" (ALO) in the following clinical settings: as an isolated condition (3 patients), idiopathic blepharospasm (BSP, 20 patients, including 4 familial cases), progressive supranuclear palsy (PSP, 7 patients), and dystonic parkinsonian syndrome (2 patients) . Twenty-nine patients treated with botulinum toxin into the orbicularis oculi muscle were rated before and after treatment and 83% of the patients improved on a clinical scale . Best results were obtained with injections directed toward the junction of the preseptal and pretarsal parts of the palpebral orbicularis oculi . Several patients also improved on anticholinergic drugs . Besides medical treatment, lid crutches, in conjunction with botulinum toxin injections, were useful in some patients . ALO is not a true apraxia; it constitutes an eyelid dystonia as shown by its clinical and electrophysiological features as well as pharmacological reactions and is encountered in a clinical spectrum ranging from an isolated form to predominant BSP . It was an important cause of treatment failures in botulinum toxin-treated BSP but by modifying our injection strategy and by adding anticholinergic drugs and also lid crutches, we obtained a good functional benefit.

Mov Disord, 1994 Nov, 9(6), 601 - 9
Botulinum toxin A injections for the treatment of hand tremors; Trosch RM et al.; We conducted an open-label study to determine the utility of treating severe hand tremors with intramuscular injections of botulinum toxin (BTX) in forearm and arm muscles in 26 patients, 12 with Parkinson's disease (PD) and 14 with essential tremor (ET) . The effect after 6 weeks for each patient was evaluated using two clinical rating scales, subjective evaluations of functional improvement and global disability, measures of weakness, and computer-assisted quantitative assessments of tremor . Although none of the clinical scores averaged > 3/4 point change, statistical significance was found on comparison of pre- and postinjection scores in the Webster Tremor and Global Disability Scales in the ET patients . Similarly, although average tremor amplitudes decreased by no more than 25% by quantitative analysis, amplitude decrease significantly correlated with patient subjective assessment in ET . In only two of 12 PD (17%) and three of 14 ET patients (21%) were major quantitative changes in tremor amplitude (> 50% reduction) found after BTX injections . Nevertheless, 10 patients (38%; five PD and five ET) reported moderate to marked subjective improvement in functional benefit after BTX . These findings suggest that although there were no major changes in clinical ratings or objective measurements, BTX injections may subjectively improve tremor in some patients, particularly those with ET.

Aust N Z J Ophthalmol, 1994 Nov, 22(4), 255 - 60
Efficacy and side effects of botulinum toxin treatment for blepharospasm and hemifacial spasm; Price J et al.; PURPOSE: To analyse the effectiveness and rate of side effects of botulinum toxin treatment for blepharospasm and hemifacial spasm . METHODS: In a prospective trial, 81 patients with blepharospasm and 70 with hemifacial spasm were treated with botulinum toxin A in the neuroophthalmology clinic at St Vincent's Hospital, Melbourne . Some 989 treatments were given and the mean follow-up time was 28.7 months . RESULTS: The duration of action was longer for patients with hemifacial spasm than for those with blepharospasm (median 12.0 weeks compared with 7.0 weeks, P < 0.0001) . There was no change in the duration of effect over time with repeated treatments of the same dose (F = 0.4, P > 0.05) . Once an effective dosage was reached, increasing the dose further did not prolong the duration of effect . There were no systemic side effects, but there were a number of local transient side effects . The most significant side effect was ptosis, which occurred in 12% of treatments given to those patients with blepharospasm and hemifacial spasm . CONCLUSION: Botulinum toxin is an effective treatment for blepharospasm and hemifacial spasm, but there are a number of side effects, the most significant being ptosis.

FEBS Lett, 1994 Oct 31, 354(1), 1 - 6
Families of zinc metalloproteases; Hooper NM; A scheme based on the zinc binding site {1992, FEBS Lett . 312, 110-114} has been extended to classify zinc metalloproteases into distinct families . The gluzincins, defined by the HEXXH motif and a glutamic acid as the third zinc ligand, include the thermolysin, endopeptidase-24.11, aminopeptidase, angiotensin converting enzyme, endopeptidase-24.15, and tetanus and botulinum neurotoxin families . The metzincins, defined by the HEXXH motif, a histidine as the third zinc ligand and a Met-turn, include the astacin, serralysin, reprolysin and matrixin families . The inverted zincin motif, HXXEH, defines the inverzincin family of insulin-degrading enzymes, the HXXE motif defines the carboxypeptidase family, and the HXH motif DD-carboxypeptidase.

Arch Ophthalmol, 1994 Oct, 112(10), 1320 - 4
Treatment of acquired nystagmus with botulinum neurotoxin A; Repka MX et al.; OBJECTIVE: Acquired nystagmus may cause oscillopsia and in some cases decreased visual acuity . Such symptoms may be debilitating . We evaluated the efficacy of retrobulbar botulinum neurotoxin A in the visual rehabilitation of patients with acquired symptomatic nystagmus . PATIENTS: Adults with acquired nystagmus from multiple sclerosis or brain-stem hemorrhage were recruited for this treatment study . Eligible patients were unable to perform visual tasks that they had performed prior to the onset of the nystagmus . DESIGN AND INTERVENTION: A prospective study evaluated the results of the retrobulbar injection of 25 to 30 U of botulinum neurotoxin A . Patients underwent testing of visual function, including eye movement recordings before and after initial injections . Patients were followed up for changes in their visual function for at least 6 months following the last injection . RESULTS: Six patients (nine eyes) with acquired nystagmus were treated with a series of 17 injections of retrobulbar botulinum neurotoxin A . Each patient had subjective and objective improvement in distance visual acuity following the injection . A reduction in the amplitude of the nystagmus was seen following each of the injections, but the frequency of the nystagmus was generally unchanged . Visual improvement usually lasted no more than 8 weeks . However, improvement persisted for 6 months after injection in two patients with oculopalatal myoclonus . CONCLUSION: Botulinum neurotoxin A transiently improves the visual function of patients with acquired nystagmus . For patients with oculopalatal myoclonus the improvement seems to last longer, about 6 months in two patients.

Neurology, 1994 Oct, 44(10), 1861 - 4
Modulation of parkinsonian tremor by radial nerve palsy; Pullman SL et al.; We analyzed rest and postural hand tremors in a Parkinson's disease patient who developed and recovered from a right radial nerve palsy at the spiral groove, and found that, despite complete paralysis of all extensors below the elbow, tremor frequencies remained unchanged while tremor amplitudes actually increased . This provides compelling evidence for a central generation of parkinsonian tremor frequency that is not influenced by the effects of peripheral modulation . In addition, the increase in tremor amplitudes may be due to disinhibited flexor activity caused by normally operating spinal segmental mechanisms interacting with central tremor generators programmed to alternate between antagonist muscles . Peripheral treatment of tremors--with muscle paralysis or botulinum toxin, for example--therefore may not be effective in stopping tremor oscillations in Parkinson's disease and may even worsen tremor amplitudes if all antagonists of a tremoring joint are not treated equally.

Electromyogr Clin Neurophysiol, 1994 Oct-Nov, 34(7), 403 - 7
Electromyographic assessment of spasmodic dysphonia patients prior to botulinum toxin injection; Rodriquez AA et al.; Electromyographic (EMG) evidence of inappropriate muscle activity (IMA) in the cricothyroid (CT) and vocalis (V) (thyroarytenoid) muscles was correlated with clinical voice measures in 32 patients with spasmodic dysphonia (SD) . Subjective voice rating and quantified fluency and laryngeal diadochokinesis measures were obtained prior to botulinum toxin (Botox) injection into the V muscles . Pre-Botox EMG was performed using a monopolar needle electrode . Each muscle was sequentially examined at rest, during vocal click, scale, sustained "E" at different pitches, and repeated "E" voicings for brief periods . A three point EMG severity scale was used to grade the amount of IMA seen in each muscle . EMG evaluation showed no evidence of lower motor neuron involvement but did reveal IMA in 81.3% of the subjects . There were no significant correlations for the patients between different EMG-based IMA severity scales and the measures of voice quality and sound production . EMG did discriminate between predominantly adductor and abductor SD pattern types, but could not correctly differentiate a mixed SD group . Those patients with adductor SD displayed IMA in the V and CT muscles, while those with abductor SD displayed more IMA in the CT than the V muscles . Sequential EMG assessment of CT and V IMA in SD did not predict clinical severity or outcome following Botox injection into the V muscles.

Pain, 1994 Oct, 59(1), 65 - 9
Botulinum toxin in the treatment of myofascial pain syndrome; Cheshire WP et al.; Six patients with chronic myofascial pain syndrome involving cervical paraspinal and shoulder girdle muscles received trigger point injections of botulinum toxin type A (Botox) or saline in a randomized, double-blind, placebo-controlled study . Four patients experienced reduction in pain of at least 30% following Botox, but not saline, injections, as measured by visual analog scales, verbal descriptors for pain intensity and unpleasantness, palpable muscle firmness, and pressure pain thresholds . Results were statistically significant . Botox, which inhibits muscle contraction by blocking the release of acetylcholine from peripheral nerves, appears to be an effective treatment for focal myofascial pain disorders.

Curr Opin Neurobiol, 1994 Oct, 4(5), 626 - 32
The molecular machinery for fast and slow neurosecretion; Martin TF; Recent studies indicate that the molecular machinery for synaptic vesicle docking and fusion consists of a triad of botulinum/tetanus neurotoxin substrates (synaptobrevin, syntaxin, SNAP-25) that are homologues of proteins required for constitutive secretion . Proposed low-affinity Ca2+ sensors that regulate exocytosis remain to be identified, although recent studies on synaptotagmin suggest that it, along with other proteins, could play this role . Regulated peptide secretion from dense-core granules has been found to utilize a similar machinery for docking/fusion, and recent studies indicate that this pathway involves a pre-docking step that is regulated by a higher affinity Ca2+ sensor.

J Neurophysiol, 1994 Oct, 72(4), 2041 - 4
Botulinum neurotoxin alters the discharge characteristics of abducens motoneurons in the alert cat; Moreno-Lopez B et al.; 1 . The effects of botulinum neurotoxin (BoTx) injected into the lateral rectus muscle were examined in alert cats by recording the extracellular activity of abducens motoneurons during spontaneous eye movements . 2 . A single high dose (3 ng/kg) of BoTx produced a complete paralysis of abduction that lasted for more than 2 mo . In addition, changes were found in the discharge pattern of abducens motoneurons . Motoneurons discharged steadily at a low firing rate (15-50 spikes/s), which in some instances showed a complete independence of eye position . Their increases in activity during ON-directed saccades were markedly reduced with respect to controls . The loss of inhibitory signals for OFF-directed saccades was even more evident . 3 . A low dose (0.3 ng/kg) of BoTx also produced a paralysis of the lateral rectus muscle that lasted for approximately 1 mo . In this case, only minor modifications in the firing characteristics of abducens motoneurons were observed . 4 . The present findings indicate that the effects of BoTx observed in the discharge pattern of abducens motoneurons might be due not only to target disconnection, but also to a central action of the neurotoxin on the motoneuron.

J Dermatol Sci, 1994 Oct, 8(2), 103 - 9
Effect of botulinum C3 exoenzyme on cell growth and cytoskeleton organization in transformed human epidermal cells in culture: a possible role for rho protein in epidermal cells; Yamamoto M et al.; We examined the role of rho gene products (rho proteins) on cell growth and cytoskeleton organization in transformed human epidermal cells in culture (HSC-1), using recombinant botulinum C3 exoenzyme which specifically ADP-ribosylates rho proteins . Incubation of HSC-1 cell lysates with C3 exoenzyme revealed a single {32P}ADP-ribosylated protein with a molecular weight of 23,000 . This protein was identified as rhoA protein by isoelectric focusing (pI 6.0) . Addition of C3 exoenzyme to the culture medium of HSC-1 cells changed the shape of HSC-1 cells to a round form with beaded processes in a time- and dose-dependent manner . Moreover, C3 treatment reduced the cell growth rate; 72-h treatment with C3 exoenzyme at 1, 3, 10, 30 and 60 micrograms/ml culture medium resulted in 9.0 +/- 1.8%, 20 +/- 2.9%, 26 +/- 2.3%, 50 +/- 1.4% and 40 +/- 2.0% inhibition of the growth rate relative to controls, respectively . Under this condition, actin stress fibers were disassembled, as revealed using fluorescent-labeled phallacidin, whereas keratin intermediate filaments were not affected, visualized by immunofluorescence using anti-keratin antibody . These results suggest that rho proteins are closely related to cell growth and that these proteins regulate, at least in part, the assembly of actin stress fibers in transformed human epidermal cells.

Electroencephalogr Clin Neurophysiol, 1994 Oct, 93(5), 325 - 9
Electromyographic single motor unit potentials after repeated botulinum toxin treatments in cervical dystonia; Odergren T et al.; Electromyographic (EMG) single motor unit potentials (MUPs) of the sternomastoid muscles (STM) were made before and after repeated treatment with botulinum type A toxin (Bx) for cervical dystonia . Post-treatment examinations were 6-25 weeks after the latest injection, when symptoms and EMG interference pattern had recurred and signs of denervation were scarce . Concentric needle EMG records of 200 motor unit potentials in 10 patients showed reduced durations and areas after treatment (P < 0.05) . Increased polyphasia or satellite potentials were not observed . Macro-EMG records of 110 MUPs in 6 patients showed reduced amplitudes and areas in the injected STM when compared to the untreated side (P < 0.05) . Fibre density was within the same range (1.0-1.2) . The results indicate that the pattern of the terminal innervation is mainly restored even after repeated Bx treatments, but the number or size of active muscle fibres within the motor unit is reduced . The clinical relapse could be due to recovery of the original nerve terminals, or to nerve sprouts closely imitating the blocked terminal nerve twigs or both.

Curr Opin Ophthalmol, 1994 Oct, 5(5), 20 - 4
Nystagmus; Spielmann A; Ophthalmologic nystagmus can be congenital and manifest/latent both of whose waveforms, compensatory mechanisms, and treatment are different . Physiologically, latent nystagmus may be reversed at will, whereas nystagmus and head nodding may be simultaneously triggered at will . Occasional oscillopsia is more frequent than is usually believed; extraretinal signals may be a reason for their absence in congenital nystagmus . Astigmatism could be the consequence of nystagmus, myopia is present in most of the cases of congenital nystagmus with blocking convergence, whereas depth-of-focus anomalies are probably due to the presence of nystagmus during its critical period of development . As for treatment, botulinum toxin injection, because of its temporary effect, is not used to treat nystagmus but to help in decision-making for surgery . Four large horizontal recti recession is the last-resort surgery which may be combined with other surgical procedures and can give unexpected results . Artificial divergence surgery by contrast is one of the most valuable procedures used in congenital nystagmus.

FEBS Lett, 1994 Sep 5, 351(2), 207 - 10
SNAP-25 is present in a SNARE complex in adrenal chromaffin cells; Roth D et al.; SNAP-25 (synaptosomal-associated protein 25 kDa) is a target for botulinum neurotoxins A and E, which both inhibit neurotransmitter release, and was recently identified together with syntaxin and synaptobrevin as receptors for NSF and alpha-SNAP . We show that SNAP-25 was enriched in the microsomal fraction from adrenal medulla, although the level of SNAP-25 in adrenal medullary microsomes was about 20-fold less than in brain microsomes . Immunocytochemistry confirmed the presence of SNAP-25 in cultured chromaffin cells and showed plasma membrane staining . Using immunoprecipitation, we found that SNAP-25 was present in a complex with syntaxin, synaptobrevin, synaptotagmin, NSF, alpha-SNAP and other unidentified polypeptides . These data indicate that SNAP-25 in chromaffin cells is present in a complex similar to that identified in brain.

Toxicol Appl Pharmacol, 1994 Sep, 128(1), 69 - 77
Measurement of botulinum toxin activity: evaluation of the lethality assay; Pearce LB et al.; The use of the mouse lethality assay for the estimation of the biologic activity of botulinum toxin was evaluated . The relationship between the number of animals, number of doses, and duration of the assay used to estimate the LD50 and the precision of the assay was investigated . The results of these studies demonstrated that the LD50 for botulinum toxin can be estimated with a high degree of precision (+/- 5%) . The precision of the assay is not increased by using more than a 5-dose 50-animal assay or extending the duration of the assay beyond 72 hr . Estimates of the LD50 obtained at 48 hr were only slightly less precise but underestimated the LD50 by 15% . Analysis of the commercially available preparations of botulinum toxin with the mouse LD50 assay revealed significant discrepancies between the units of toxin in these preparations . In addition, a 2.67-fold difference in the relative potency of the two preparations of botulinum A toxin was observed using a regional chemodenervation assay that measures paralysis . The mouse LD50 assay could not detect this large difference in the potency of the two approved clinical preparations of botulinum toxin . The results of these studies demonstrate that although the mouse LD50 assay can be used to estimate the number of units of botulinum toxin with a high degree of precision this assay alone is not an adequate method for assessing the preclinical biological potency of botulinum toxin.

J Immunol, 1994 Sep 1, 153(5), 1998 - 2003
Epstein-Barr virus induces actin polymerization in human B cells; Melamed I et al.; The EBV selectively infects human and primate lymphocytes . This selective tropism occurs as a result of live virus infection through permissive membrane receptors . Once EBV has entered the cell, it induces proliferation and immortalization of these cells . The mechanism of EBV infection, however, remains largely unknown . We demonstrate here that a transforming strain, but not a nontransforming strain, of EBV stimulates the conversion of globular actin (G-actin) to filamentous actin (F-actin), a process that has been associated with activation and transformation of other cell types . Preincubation of B cells with botulinum C2 toxin or cytochalasin, which block the conversion of G-actin to F-actin, resulted in the inhibition of EBV-induced proliferation . These findings indicate that actin rearrangement is essential for infection of B cells by EBV.

Headache, 1994 Sep, 34(8), 458 - 62
Tension headache: botulinum toxin paralysis of temporal muscles; Zwart JA et al.; The pathogenetic mechanism of tension headache (TH) is still unknown . The role of pericranial muscle tension in TH is also enigmatic . To evaluate this factor in chronic TH, pericranial muscles were paralysed in 6 chronic TH patients, using botulinum toxin . All patients fulfilled the IHS criteria of chronic TH associated with involvement of the pericranial muscles, but not the current criteria for cervicogenic headache . The patients were followed-up regularly with evaluation of the paralysis, changes in pain intensity, and pressure pain threshold measurements . We primarily only injected the temporal muscle on the one side, using the other side as a control . Contralateral muscles were in some cases injected at a later stage . In our study, we did not find any significant reduction in pain intensity, as measured by the visual analogue scale, nor any changes in pressure pain threshold, as measured by an algometer . On the basis of our observations, we conclude that muscle tension in these muscles possibly plays a minor role in the genesis of chronic TH . In our study, however, we have only treated a limited number of patients, and only one pericranial muscle has been injected systematically . Further studies of various neck/posterior head muscles ought to be performed in order to further evaluate a possible effect of tension in the pericranial musculature in producing this type of pain.

Gut, 1994 Sep, 35(9), 1319 - 21
Intrasphincteric injection of botulinum toxin for suspected sphincter of Oddi dysfunction; Pasricha PJ et al.; Botulinum toxin is a potent inhibitor of the release of acetylcholine from nerve endings . It has previously been shown that it can effectively reduce lower oesophageal sphincter pressures both in animals and humans with achalasia . This study examined the hypothesis that locally injected botulinum toxin could also reduce sphincter of Oddi pressure in patients with sphincter of Oddi dysfunction . Two patients with postcholecystectomy pain syndrome were diagnosed with sphincter of Oddi dysfunction (by biliary manometry in one patient and by hepatobiliary scanning criteria in the other) . Botulinum toxin was injected into the sphincter of Oddi, by a sclerotherapy needle passed through a duodenoscope . In the first patient, intrasphincteric injection of botulinum toxin reduced sphincter pressure by about 50%, an effect that was sustained for at least four months . In the second patient, intrasphincteric injection caused about a 50% improvement in bile flow, with normalisation of scintigraphy . Neither patient showed any sustained improvement in pain despite these objective findings . Both patients eventually had endoscopic sphincterotomy, which also did not result in symptomatic improvement in either patient . No side effects were seen . Intrasphincteric botulinum toxin is a simple and effective means of lowering sphincter of Oddi pressure . This technique has potential for being useful clinically.

Ophthal Plast Reconstr Surg, 1994 Sep, 10(3), 193 - 4
Oculinum injection-resistant blepharospasm in young patients; Gausas RE et al.; Botulinum toxin has recently been used as a nonsurgical treatment for blepharospasm and other facial dyskinesias . This report describes four patients between the ages of 32 and 37 years who failed to respond to botulinum injections for severe blepharospasm . Other than age, no other features could be identified in these patients that would differentiate this group . Particularly early age of onset for essential blepharospasm might be an indicator of failure to respond to the injections.

Neuromuscul Disord, 1994 Sep-Nov, 4(5-6), 489 - 96
Calcitonin gene-related peptide-like immunoreactivity, in botulinum toxin-paralysed rat muscles; Hassan SM et al.; Changes in calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) at the motor endplates of botulinum toxin-paralysed rat muscles were investigated using immunohistochemistry . One day following toxin injection, a dramatic increase in CGRP-LI was detected at the motor endplates and within preterminal axons of the soleus and gastrocnemius muscles . The upregulation of CGRP-LI persisted throughout the period during which muscle fibres were paralysed and new neuromuscular junctions were being formed by the growing sprouts . Decline of CGRP-LI at the motor endplates coincided with clinical recovery . Both up- and down-regulation of CGRP-LI took place earlier in the soleus than in the gastrocnemius muscle . Up-regulation of CGRP-LI was also detected in a subpopulation of motor axons in the sciatic nerves and in the spinal motor neurons innervating the paralysed muscles . These results indicate that levels of CGRP are regulated, at least partly, by changes in the target innervation . They also suggest an important role for CGRP in the regenerative processes following muscle paralysis.

J Pediatr Ophthalmol Strabismus, 1994 Sep-Oct, 31(5), 283 - 6
Results of a prospective randomized trial of botulinum toxin therapy in acute unilateral sixth nerve palsy; Lee J et al.; Forty-seven patients entered a prospective randomized trial to assess the effect of early botulinum neurotoxin A treatment to the ipsilateral antagonist medial rectus on the ultimate recovery rate of acute unilateral sixth nerve palsy . Twenty-two patients received injections and 25 acted as controls . The overall etiologies were microvascular (72.3%), unknown (17%), multiple sclerosis (6%), and one case each of central nervous system (CNS) sarcoidosis and basilar artery ectasia . Eighty-three percent of the patients entered the trial within 2 weeks of the onset of symptoms and 95.7% within 3 weeks . The controls had a final recovery rate of 20/25 (80%), and the injected group had a final recovery rate of 19/22 (86%) . No serious side effects were encountered . We conclude that there is no evidence for a prophylactic effect of botulinum toxin in the group that we have studied.

Nurse Pract, 1994 Sep, 19(9), 67 - 73
Spasmodic dysphonia: new diagnosis and treatment opportunities; Cook MJ et al.; The ability to communicate is a basic need . Spasmodic dysphonia, a rare dystonia focal to the larynx, affects the vocal muscles resulting in abnormal sound and speech production . The voice may become hoarse, strained, strangled, tremulous, whispered, or aphonic . Lack of appropriate diagnosis along with the continued use of an abnormal voice often results in frustration, anxiety, and depression . Improved diagnostic ability and unique treatment with botulinum toxin injection (Botox) are now available to patients in several large medical centers across the United States and Canada . A knowledgeable and observant health care provider can alert patients and families to the possibility of treatment for a chronic voice disturbance.

J Med Assoc Thai, 1994 Sep, 77(9), 464 - 70
Botulinum A toxin treatment in spasmodic torticollis: report of 56 patients; Poungvarin N et al.; Botulinum A toxin injection is the most recent and effective treatment of various movement disorders especially focal dystonia . Spasmodic torticollis is one focal dystonia which responds poorly to both medication and surgery . Botulinum A toxin injection has been adopted as a treatment procedure at the Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand since 1989 (before the American Food and Drug Administration approval) as a research protocal for Thai patients . This report is the first ever study of this treatment for Thai patients with spasmodic torticollis . Fifty six spasmodic torticollis patients who had been treated with botulinum A toxin injection at the Movement Disorder Clinic, Siriraj Hospital were analysed . Thirty six patients were male and the male to female ratio was 1.8:1 . Most of the patients (76.79 per cent) were aged between 20-49 years and half of them were from Bangkok . Twelve patients (21.43 per cent) were classified as simple torticollis, 35 patients (62.5 per cent) were combined torticollis, 7 patients (12.5 per cent) were retrocollis, and 2 patients (3.57 per cent) were lateral collis . Three patients had generalised dystonia and 2 patients had segmental dystonia . Duration of suffering in each patient ranged from 1 month to 25 years with the mean duration of 3.70 (S.D . 5.09) years . Only four patients (7.14 per cent) refused botulinum A toxin injection due to their mild symptoms . The remaining 52 patients were given botulinum A toxin injection of 30-120 international units into the most overactive group of muscles which were responsible for abnormal neck posture (mainly sternocleidomastoid and splenius capitis) . Eight patients (15.38 per cent) were lost to follow-up.(ABSTRACT TRUNCATED AT 250 WORDS)

Dev Comp Immunol, 1994 Sep-Oct, 18(5), 389 - 95
Inhibition of phagocytosis by rainbow trout (Oncorhynchus mykiss) macrophages by botulinum C2 toxin and its trypsinized component II; Kodama H et al.; Botulinum C2 toxin (C2T) is composed of two nonlinked protein components, components I and II . The toxin reconstituted with component I and trypsinized component II inhibited phagocytosis of rainbow trout (Oncorhynchus mykiss) and mouse macrophages . Inhibition in both cell types was observed over a range of toxin concentrations that were not toxic to the cells . Because cytoplasmic action of rainbow trout macrophages is ADP-ribosylated by component I, the inhibition of phagocytosis in trout cells by C2T is probably due to inactivation of cytoplasmic actin . Moreover, phagocytosis by trout cells was also inhibited in a dose-dependent manner by trypsinized component II alone, and did not cause cell death . The present results show that the macrophages of aquatic vertebrates are susceptible to C2T, and that trypsinized component II elicits a novel biological activity by binding to the cell membrane of the macrophages.

J Biol Chem, 1994 Aug 12, 269(32), 20213 - 6
Botulinum G neurotoxin cleaves VAMP/synaptobrevin at a single Ala-Ala peptide bond; Schiavo G et al.; Similarly to other serotypes, botulinum neurotoxin serotype G (BoNT/G) contains the zinc binding motif of zinc endopeptidases . Highly purified preparations of BoNT/G show a zinc-dependent protease activity specific for VAMP/synaptobrevin, a membrane protein of synaptic vesicles . The two neuronal VAMP isoforms are cleaved with similar rates at one Ala-Ala peptide bond present in the same region, out of the several such peptide bonds present in their sequences . This site of cleavage is unique among the eight clostridial neurotoxins . VAMP proteolysis is displayed only after reduction of the single interchain disulfide bond present in the toxin, and it is inhibited by EDTA, o-phenanthroline and captopril.

Ann Neurol, 1994 Aug, 36(2), 129 - 41
Treatment of abnormal eye movements that impair vision: strategies based on current concepts of physiology and pharmacology; Leigh RJ et al.; Certain abnormal eye movements, especially pathological nystagmus, degrade vision and cause illusory motion of the seen environment . These symptoms are due to excessive movement of images of stationary objects on the retina . Recently, the pathophysiology underlying several types of nystagmus and saccadic oscillations was better defined by the development of animal models and by experimental pharmacological studies . Despite this, few reliable therapies are currently available for these abnormal eye movements . In clinical studies, a number of drugs reportedly helped individual patients, but few drugs have been subjected to double-blind trials . An alternative approach to pharmacological suppression of abnormal eye movements is optical stabilization of images on the retina, which is helpful in selected patients . Weakening of the extraocular muscles, using botulinum toxin or surgery, is prone to cause diplopia and may induce plastic-adaptive changes that render the effect temporary . In some patients, treatment of an underlying condition, such as the Arnold-Chiari malformation, reduces nystagmus and improves vision . There is a need for multicenter trials to evaluate systematically potential treatments of abnormal eye movements that impair vision.

Curr Opin Neurol, 1994 Aug, 7(4), 358 - 66
Botulinum toxin in movement disorders; Jankovic J; The most potent biologic toxin, botulinum toxin (BTX), has become a powerful therapeutic tool in the treatment of a variety of neurologic, ophthalmic, and other disorders manifested by abnormal, excessive, or inappropriate muscle contractions . This review focuses on the use of BTX in the treatment of dystonia and other movement disorders . The therapeutic application of BTX, however, extends beyond movement disorders; chemodenervation with BTX has been found to ameliorate spasticity, rigidity, spastic bladder, achalasia, and even some cosmetic conditions . In addition to describing its therapeutic effects, this article also reviews recent advances in the understanding of the molecular and cellular mechanisms of BTX . Few therapeutic agents have been better understood in terms of their mechanism of action or have had greater impact on patients' functioning than BTX . BTX-A has been used in nearly all clinical trials . Blocking anti-BTX-A antibodies have been detected in about 5% of patients chronically treated with this type of BTX . Patients who develop immunoresistance to BTX-A may benefit from other serotypes of BTX, such as BTX-B and -F, currently undergoing clinical trials.

Plast Reconstr Surg, 1994 Jul, 94(1), 94 - 9
Botulinum toxin A for hyperkinetic facial lines: results of a double-blind, placebo-controlled study; Keen M et al.; Previous work on patients with muscular dystonia has shown that small intramuscular doses of botulinum toxin A eliminated hyperkinetic facial lines for approximately 6 months . The purpose of this study was to determine the efficacy of botulinum toxin A injections in eliminating facial wrinkles in aesthetic surgery patients who do not have muscular dystonia . Eleven healthy subjects were studied in a double-blind fashion . On both sides of the face, 0.2 cc of either normal saline or botulinum toxin A was injected into the forehead or into the periorbital wrinkles (crow's feet) . Documentation of results was made by photographs taken of the patients during repose and during facial animation before and after injection . Assessment of facial wrinkles was done from a grading system in which the patient and the facial plastic surgeon were asked to judge the severity of the wrinkles on a scale from 0 to 3, with 0 reflecting no facial wrinkles and 3 reflecting severe facial wrinkling . Nine of 11 subjects injected with botulinum toxin A noted a significant improvement in the severity of their facial wrinkles in comparison with the side of the face injected with saline, with a rating improvement of 2 points . Two of 11 subjects noted a moderate improvement, with a rating improvement of 1 point . No patient injected with saline reported an improvement in the severity of the facial wrinkles on the control side . There were no serious complications . Botulinum toxin A is an efficacious method of nonsurgically eliminating facial wrinkles and may play a role in the cosmetic enhancement of the aging face.

Muscle Nerve, 1994 Jul, 17(7), 779 - 84
Botulinum toxin converts muscle acetylcholine receptors from adult to embryonic type; Koltgen D et al.; To assess the postsynaptic consequences of botulinum toxin injection into muscle we characterized the nicotinic acetylcholine receptor (nAChR) with the patch clamp technique, using adult mouse muscle after destruction of the nerve ending and after treatment with botulinum toxin (BoTX) . In both, embryonic channels with a conductance of 30 and 34 pS could be identified, whereas on adult control muscle nAChR channels had a conductance of 48 pS . The mean open times were 1.2 ms for the channels in control, 2.5 ms in denervated and 2.4 ms in BoTX-treated muscle . The dose-response curves of the maximal acetylcholine-elicited currents showed a Km of 60 mumol/L for denervated, 70 mumol/L for BoTX-treated, and 100 mumol/L for control muscle . Destruction of the nerve ending and inhibition of acetylcholine release by BoTX has the same effect as far as the increase of sensitivity of the muscle to acetylcholine is concerned . In contrast to single-fiber EMG findings in patients treated for focal dystonia no distant changes could be found in the control muscle of the BoTX-treated animals.

Mov Disord, 1994 Jul, 9(4), 395 - 402
Clinical and electromyographic features of levator palpebrae superioris muscle dysfunction in involuntary eyelid closure; Aramideh M et al.; We report on five patients with involuntary eyelid closure, diagnosed as blepharospasm and referred to use for treatment with botulinum A toxin . Synchronous electromyographic (EMG) recording was performed from the levator palpebrae superioris (LP) and the orbicularis oculi (OO) muscles . In the first two cases, EMG registration showed alternating, semirhythmic dystonic activities in both the LP and OO, clinically perceptible as "flickering" of the eyelids . While the eyelids were lowered, one of them also showed involuntary upper eyelid tractions due to dystonic activities of LP . In the third patient, EMG patterns were characterized by a gradual decrease in the level of LP activity, followed by the contraction of OO, which facilitated the return of LP to its tonic activity, termed "postinhibition potentiation" of LP . In the fourth patient, EMG recording showed involuntary inhibition of LP in combination with blepharospasm . Involuntary closure of the eyelids in the fifth patient was caused by short or prolonged periods of involuntary LP inhibition, whereas OO activity remained normal . Our results provide further evidence that LP muscle activities are regulated by burst-tonic motoneurons, and we suggest that these motoneurons, and/or the input signals regulating their activities, can be involved independently in a pathological process . Clinical symptoms are discussed that may be helpful to recognize those cases with LP motor dysfunction, whether or not accompanied by OO activity disorders . Because the term blepharospasm indicates an abnormal motor function of OO, we propose "blepharospasm-plus" to designate those cases with a combined motor dysfunction of LP and OO muscles.

Neurosurgery, 1994 Jul, 35(1), 58 - 62; discussion 62-3
Selective peripheral denervation for the treatment of spasmodic torticollis; Braun V et al.; The results of selective peripheral denervation in 50 patients with spasmodic torticollis are presented . Of our patients, 76% reported a significant improvement or disappearance of their dystonia . The mean follow-up is 25 months . There were no major side effects . We recommend the procedure to patients who primarily have responded to botulinum toxin therapy and had become secondary nonresponders or to those refusing further injections while still responding . The results are much less promising in patients who are primary nonresponders to botulinum toxin . Some remarkable histological findings are presented . The posterior branches of the cervical roots frequently showed signs of severe compression neuropathy . In three cases, a functional motor nerve regeneration was proved . Among all surgical options, selective peripheral denervation provides the best result and has the fewest side effects.

J Pediatr Ophthalmol Strabismus, 1994 Jul-Aug, 31(4), 214 - 9
Observations on bilateral simultaneous botulinum toxin injection in infantile esotropia; McNeer KW et al.; Botulinum toxin (BoTX) is an effective pharmacological alternative to the surgical management of strabismus in adults . In a previous study, we found that concurrent bilateral medial rectus muscle BoTX injections for infantile esotropia could produce stable ocular realignment without significant risk . No other investigators have used bilateral simultaneous BoTX injection . We report the results of bilateral simultaneous medial rectus BoTX injection in 57 infantile esotropia patients followed for a minimum of 12 months . The esotropic angle in 27 infantile esotropia patients under 12 months of age was reduced from 43 +/- 12 to 1 +/- 2 prism diopters . The esotropic angle in 30 infantile esotropia patients at a mean age of 24 months was reduced from 31 +/- 12 to 2 +/- 3 delta . Other variables including late onset of hyperopic refractive errors, dissociated vertical deviations, overacting oblique muscles, and consecutive exodeviations are evaluated . We regard BoTX as reliable therapy in infantile esotropia.

Tidsskr Nor Laegeforen, 1994 Jun 20, 114(16), 1830 - 1
{Treatment of writer's cramp with botulinum toxin}; Amthor KF et al.; Writer's cramp is a focal dystonia that is resistant to most treatment regimens . This article describes two patients with writer's cramp who were successfully treated with botulinum toxin injections into the forearm muscles . Transient weakness due to the effect of botulinum toxin was registered in the injected muscles . No other side effects were observed . The beneficial effect of a single injection lasted from three to four months and could be reproduced by repeated injections over a three year follow-up period . We conclude that botulinum toxin injections may provide safe and effective treatment in selected patients with writer's cramp.

J Neurosci Res, 1994 Jun 15, 38(3), 294 - 9
Neural regulation of muscle acetylcholinesterase is exerted on the level of its mRNA; Cresnar B et al.; In rat muscles, AChE activity drops rapidly after denervation, and the patterns of AChE molecular forms in slow and fast muscles differ considerably . Both observations imply that muscle AChE is regulated by the motor nerve . In order to obtain a better insight into the underlying mechanism, AChE regulation in rat muscles was examined on the level of its catalytic subunit mRNA using northern blot analysis . The level of two AChE transcripts (2.4 and 3.2 kb) was much higher in the fast sternomastoid (STM) than in the slow soleus muscle, which explains the difference in AChE activity between the two types of muscles . Expression of AChE mRNA in the extrajunctional region of STM muscle is fairly high so that little difference in the level of AChE mRNAs was observed in comparison to the region rich in the neuromuscular junctions . This indicates that very high AChE activity in the neuromuscular junctions is achieved by unique posttranslational modifications and cellular processing of AChE enhancing stability of the junctional in comparison to the extrajunctional AChE . Denervation as well as botulinum toxin evoked paralysis of STM muscle caused rapid decline of AChE transcripts to almost undetectable levels both in the junctional and extrajunctional regions . The low level of AChE mRNA is therefore largely responsible for low AChE activity in denervated rat muscles . It seems that either muscle activity and/or quantal ACh release enhance the level of AChE mRNA in the junctional as well as extrajunctional regions . In rat muscles, extrajunctional mRNA level of the catalytic subunit of AChE is neurally regulated in exact opposite fashion from that of acetylcholine receptor subunits.

Muscle Nerve, 1994 Jun, 17(6), 623 - 31
Elimination of superfluous neuromuscular junctions in rat calf muscles recovering from botulinum toxin-induced paralysis; Hassan SM et al.; In order to determine the fate of the superfluous neuromuscular junctions (NMJs) formed during the course of botulinum toxin (BoTx)-induced paralysis, we have quantified the change in the total length of the nerve muscle contact area(s) following BoTx injection into rat calf muscles . The results indicate that: (1) at least some of the superfluous NMJs are eliminated following muscle recovery; (2) synapse elimination is a slow process, as 4 months after recovery it was not yet complete; (3) muscles with different content of type I and II fibers follow a different time course during synapse formation and elimination . We further investigated the possibility that the neural cell adhesion molecule (NCAM) would be the element whose loss from the NMJ might play a role in synapse elimination . Using immunofluorescence and immunoelectron microscopy we show that NCAM is exclusively localized between nerve terminals and Schwann cells and not between nerve terminals and muscle . This localization was maintained throughout paralysis and following recovery, suggesting that NCAM does not play a role in synapse elimination.

Laryngoscope, 1994 Jun, 104(6 Pt 1), 656 - 62
Anatomic considerations in botulinum toxin type A therapy for spasmodic dysphonia; Castellanos PF et al.; Chemodenervation by injection of botulinum toxin type A into the vocal fold(s) has become the preferred treatment for patients with adductor spasmodic dysphonia . Injection may be done either perorally or transcutaneously; each method has its advocates and advantages . The authors have used the transcutaneous transcricothyroid membrane route exclusively with satisfactory results in more than 50 patients . Temporary breathliness and aspiration are common . The preferred injection site should be as close as possible to the motor end plates of the affected muscle . The thyroarytenoid muscle end plates are distributed throughout the muscle, whereas in the lateral cricoarytenoid muscle they are located in band in the center of the muscle . The transcutaneous injection site is below and posterior to the midpoint of the vibrating vocal fold as visualized by indirect laryngoscopy . The proximity of this site to the lateral cricoarytenoid muscle suggests that postinjection breathiness and aspiration may be related to spread of botulinum toxin type A to the lateral cricoarytenoid muscle . However, it is likely that thyroarytenoid muscle paresis is mainly responsible for this side effect and that the rapid clearing of the breathy dysphonia in the face of prolonged relief of spasmodic dysphonia symptoms suggests the action of an adaptive neural response, such as axonal sprouting . Further research of this subject is warranted.

J Neurocytol, 1994 Jun, 23(6), 354 - 63
GAP-43 and p75NGFR immunoreactivity in presynaptic cells following neuromuscular blockade by botulinum toxin in rat; Hassan SM et al.; Peripheral nerve lesion results in changes in protein expression by neurons and denervated Schwann cells . In the present study we have addressed the question whether similar changes take place following functional denervation . Using immunohistochemistry and immunoelectron microscopy we examined changes in growth-associated protein (GAP-43) and low-affinity nerve growth factor receptor (p75NGFR) in rat gastrocnemius muscle following botulinum toxin-induced paralysis . GAP-43 and p75NGFR were selected because they are not expressed by mature intact motor neurons or Schwann cells, but are expressed following nerve lesion in both motor neurons and denervated Schwann cells . In control muscle, GAP-43 and p75NGFR immunoreactivity was seen only in nerve fibres near blood vessels . Two weeks after toxin injection, GAP-43 immunoreactivity could be seen at the motor endplates and in axons . Intensity of staining increased with longer survival and reached a peak between 4 and 8 weeks post-injection . Ultrastructurally, GAP-43 immunoreactivity was confined to nerve terminals and axons, whereas Schwann cells remained negative . Immunostaining for p75NGFR also increased following toxin injection and was detected in some terminal Schwann cells and in perineurial cells of small nerve fascicles near the paralyzed target cells, but not in axons . These results show that changes in expression of GAP-43 in motor neurons following functional denervation closely resemble the changes following anatomical interruption of nerve-muscle contact . GAP-43 was not expressed in Schwann cells, indicating that its upregulation in these cells is induced by loss of axonal contact or nerve degeneration products . There is no support for a role of p75NGFR in incorporation of neurotrophins in axons . The restriction of p75NGFR expression to terminal Schwann cells and perineurial cells in close proximity to the paralyzed target suggests a role for a target-derived signal or, alternatively, macrophages in eliciting this expression.

Pediatr Neurol, 1994 Jun, 10(4), 284 - 8
Botulinum toxin A in management of cerebral palsy; Calderon-Gonzalez R et al.; The efficacy of local injection of botulinum toxin A in selected skeletal muscles to relieve muscle hypertonia and muscle contracture, and increase range of motion in children with cerebral palsy was studied in an open ABA (baseline-treatment-posttreatment phase) type of study . The first 6 months were the baseline phase, the day of injection the treatment phase, and the next 6 months the posttreatment phase . The patients acted as their own controls . Fifteen children with cerebral palsy (mean age: 6 years, 8 months) were included in the study . All had limb deformities associated with non-fixed joint contractures that had not responded to physical therapy . Clinical assessment of passive and active muscle tone was performed using a modified Ashworth scale . The range of motion to passive movement was measured with a manual goniometer . Botulinum toxin was injected directly into the muscle at several sites . The postinjection scores of muscle hypertonia were significantly lower (P < .01) and the range-of-motion values demonstrated a significant increase (P < .001) . Functional improvement was measured by decreased scissoring on standing in all 6 children with adductor muscles injected; all 6 children with knee flexor muscles injected were able to straighten the knees . The 3 children with injected gastrocnemius muscles were able to achieve heel-strike while bare-footed . The study provides evidence that the intramuscular injection of botulinum toxin A in selected skeletal muscles decreases muscle tone and contractures, and increases range of motion and motor function.

J Otolaryngol, 1994 Jun, 23(3), 160 - 4
A comparison of the efficacy of unilateral versus bilateral botulinum toxin injections in the treatment of adductor spasmodic dysphonia; Maloney AP et al.; The current treatment of choice of adductor spasmodic dysphonia due to focal dystonia is thyroarytenoid-vocalis injection of botulinum toxin type A (Botox) . Botox exerts its effect by presynaptic motor endplate blockade, preventing the release of acetylcholine and causing muscle paresis . Botox treatment protocols vary . Some centres perform unilateral injections, whereas others treat both cords . Our hypothesis is that unilateral injections may reduce the severity of whisper voice and aspiration side effects in the early two to three weeks after treatment . The purpose of this study, therefore, is to compare the efficacy of unilateral versus bilateral Botox injections in the treatment of adductor spasmodic dysphonia in terms of duration of effect versus the side effects of breathing and swallowing difficulties . This study presents data from a retrospective chart review and a prospective telephone interview of all patients receiving bilateral and unilateral Botox injections.

Eur J Biochem, 1994 Jun 1, 222(2), 325 - 33
Botulinum A and the light chain of tetanus toxins inhibit distinct stages of Mg.ATP-dependent catecholamine exocytosis from permeabilised chromaffin cells; Lawrence GW et al.; Susceptibilities of Mg.ATP-independent and Mg.ATP-requiring components of catecholamine secretion from digitonin-permeabilised chromaffin cells to inhibition by Clostridial botulinum type A and tetanus toxins were investigated . These toxins are Zn(2+)-dependent proteases which specifically cleave the 25-kDa synaptosomal-associated protein (SNAP-25) and vesicle-associated membrane protein (VAMP) II, respectively . When applied to permeabilised chromaffin cells they rapidly inhibited secretion in the presence of Mg.ATP but the catecholamine released in the absence of Mg.ATP, thought to represent fusion of primed granules, was not perturbed . The toxins can exert their effects per se in the absence of the nucleotide complex; therefore, Mg.ATP-requiring steps of secretion are implicated as roles for their targets . Primed release was lost rapidly after permeabilisation of the cells but could be maintained by including Mg.ATP during the incubation before stimulating release with Ca2+ . This ability of Mg.ATP to maintain primed release was only partially inhibited by botulinum neurotoxin A whereas it was abolished by tetanus toxin, consistent with the distinct substrates for these toxins . This study reveals a component of release within which these proteins are either resistant to cleavage by these toxins or in such a position that degradation can no longer prevent granule fusion . Differences in the steps of release at which these toxins can affect inhibition are also revealed.

Neuron, 1994 Jun, 12(6), 1269 - 79
A post-docking role for synaptobrevin in synaptic vesicle fusion; Hunt JM et al.; We have used the squid giant synapse to determine the role of synaptobrevin, integral membrane proteins of small synaptic vesicles, in neurotransmitter release . The sequence of squid synaptobrevin, deduced by cDNA cloning, is 65%-68% identical to mammalian isoforms and includes the conserved cleavage site for tetanus and botulinum B toxins . Injection of either toxin into squid nerve terminals caused a slow, irreversible inhibition of release without affecting the Ca2+ signal which triggers release . Microinjection of a recombinant protein corresponding to the cytoplasmic domain of synaptobrevin produced a more rapid and reversible inhibition of release, whereas two smaller peptide fragments were without effect . Electron microscopy of tetanus-injected terminals revealed an increased number of both docked and undocked synaptic vesicles . These data indicate that synaptobrevin participates in neurotransmitter release at a step between vesicle docking and fusion.

Arq Neuropsiquiatr, 1994 Jun, 52(2), 269 - 72
{Botulinum toxin the treatment of spasmodic torticollis: a meta-analysis}; Teixeira SC et al.; The authors made a meta-analysis of the results of botulinum toxin injection for the treatment of spasmodic torticollis . They concluded that botulinum toxin is effective for the treatment of cervical dystonia.

Proc Natl Acad Sci U S A, 1994 May 24, 91(11), 4688 - 92
Synaptobrevin/vesicle-associated membrane protein (VAMP) of Aplysia californica: structure and proteolysis by tetanus toxin and botulinal neurotoxins type D and F; Yamasaki S et al.; Synaptobrevin/vesicle-associated membrane protein (VAMP) and syntaxin are potential vesicle donor and target membrane receptors of a docking complex that requires N-ethylmaleimide-sensitive factor (NSF) and soluble NSF-attachment proteins as soluble factors for vesicle fusion with target membranes . Members of this docking complex are the target of clostridial neurotoxins that act as zinc-dependent proteases . Molecular cloning of the Aplysia californica synaptobrevin cDNA revealed a 180-residue polypeptide (M(r), 19,745) with a central transmembrane region and an atypically large C-terminal intravesicular domain . This polypeptide integrates into membranes at both the co- and posttranslational level, as shown by modification of an artificially introduced N-glycosylation site . The soluble and membrane-anchored forms of synaptobrevin are cleaved by the light chains of the botulinal toxins type D and F and by tetanus toxin involving the peptide bonds Lys49-Ile50, Gln48-Lys49, and Gln66-Phe67, respectively . The active center of teh tetanus toxin light chain was identified by site-specific mutagenesis . His233, His237, Glu234, and Glu270/271 are essential to this proteolytic activity . Modification of histidine residues resulted in loss of zinc binding, whereas a replacement of Glu234 only slightly reduced the zinc content.

Brain Res Dev Brain Res, 1994 May 13, 79(1), 39 - 46
Botulinum A toxin stimulates neurite branching in nerve-muscle cocultures; Bonner PH et al.; In addition to skeletal muscle paralysis, type A botulinum toxin commonly causes sprouting of motor axons in various experimental whole-animal systems . The use of type A botulinum toxin in clinical treatment of muscle spasm disorders is becoming increasingly popular . The eventual, unwanted return of involuntary activity in the treated muscles may be a consequence of such axon sprouting . We have developed a coculture model allowing the quantification of botulinum toxin-induced sprouting that shows promise for future studies on its mechanism and control . Chick embryo ciliary ganglion motor neurons were cocultured with chick leg muscle cells . The presence of type A botulinum toxin in the coculture medium was correlated with significantly increased branching frequency of neurites . Toxin-increased branching frequency occurred even when the neurons and muscle cells were separated from each other on the culture dishes, suggesting a presynaptic effect of toxin . Cocultures incubated in the presence of curare, a post-synaptic blocker, had control levels of neurite branching, ruling out the possibility that simple synaptic blockade causes sprouting but again supporting the hypothesis of a pre-synaptic activity of botulinum toxin.

Dev Med Child Neurol, 1994 May, 36(5), 386 - 96
Botulinum toxin in the management of the lower limb in cerebral palsy; Cosgrove AP et al.; The role of intramuscular botulinum toxin A in the treatment of 26 children with cerebral palsy was evaluated . The indication for injection was the presence of a dynamic contracture of lower-limb muscles interfering with positioning or walking . Spastic target muscles were identified by clinical examination and, in ambulant children, by gait analysis . Between 50 and 320 units of botulinum toxin were injected into each muscle group to a total dose of 100 to 400 units per child . The effects of injection were monitored by repeated clinical examination and gait analysis . There were no clinically detectable systemic side-effects, and all but one patient had a reduction in tone, which occurred within three days and persisted for two to four months . There were significant improvements in ambulatory status and in sagittal-plane kinematics . In some cases these gains persisted after the tone-reducing effects of the toxin had worn off.

Dev Med Child Neurol, 1994 May, 36(5), 379 - 85
Botulinum toxin A prevents the development of contractures in the hereditary spastic mouse; Cosgrove AP et al.; The hereditary spastic mouse was studied as a model of cerebral palsy in childhood to test the hypothesis that intramuscular botulinum toxin A would prevent the development of calf-muscle contractures . A prospective randomised controlled trial of calf injection with botulinum A compared with injection of normal saline was performed on juvenile mice . At maturity, the calf muscles of the spastic mice were 16 per cent shorter than those of their normal siblings . The calf muscles of spastic mice injected with botulinum toxin A grew to within 2 per cent of normal length . This difference in mature muscle length was highly significant.

Klin Monatsbl Augenheilkd, 1994 May, 204(5), 366 - 9
{Early surgery of convergent strabismus . Preliminary results of a series of 45 cases}; Iuvara-Bommeli A et al.; BACKGROUND: The age for strabismus surgery remains controversial and the various studies differ whether the results of residual angle and binocular vision may be better with early or late surgery . Different operating techniques for infantile esotropia have been proposed: recession of both medial recti, combined surgery on horizontal muscles, posterior fixation on one or both eyes, posterior fixation with simultaneous recession, botulin injection . PATIENTS: Our study presents 45 children with congenital esotropia operated at a mean age of 18.5 months . The surgical technique consisted of posterior fixation of both medial recti with or without simultaneous recession.

Mov Disord, 1994 May, 9(3), 347 - 9
Botulinum toxin in the treatment of dystonic tics; Jankovic J; Botulinum toxin (BTX) injections provide effective treatment for a variety of disorders manifested by inappropriate muscle contractions, but its efficacy in the treatment of tics has not been previously studied . Ten male patients 13-53 years of age who were diagnosed with Tourette's syndrome manifested by disabling focal tics were included in this pilot study . Five patients had frequent blinking and blepharospasm, rendering them "blind," and five patients had severe and painful dystonic tics involving their neck muscles . All 10 patients experienced moderate to marked improvement in the intensity and frequency of tics after BTX injections into the involved muscles . Patients in whom premonitory urges preceded their tics noted marked lessening of these sensory symptoms . The benefit lasted 2-20 weeks after injections . There were no serious complications, except for transient ptosis in two and neck pain, stiffness, or weakness in three patients . BTX injections appear to be safe and effective treatment for patients with focal dystonic tics . The treatment ameliorates not only involuntary movements but also the premonitory sensory component associated with some tics.

Mov Disord, 1994 May, 9(3), 318 - 20
Adductor laryngeal breathing dystonia in a patient with lubag (X-linked dystonia-Parkinsonism syndrome); Lew MF et al.; We report a patient with Lubag (X-linked dystonia-parkinsonism) who presented with severe respiratory stridor from adductor laryngeal breathing dystonia . Emergency tracheostomy was necessary, and subsequent laryngeal injection with botulinum toxin led to worsening aspiration . Botulinum toxin injection for severe lingual dystonia was successful.

J Laryngol Otol, 1994 May, 108(5), 380 - 2
Management of middle ear myoclonus; Badia L et al.; Tinnitus produced by synchronous repetitive contraction of the middle ear muscles (middle ear myoclonus) is a rare condition . We present six cases of middle ear myoclonus in whom different management regimes were successful . In two patients, the tinnitus was controlled by conservative measures . In one patient, whose tinnitus was associated with blepharospasm, significant improvement occurred following botulinum toxin injection into the ipsilateral orbicularis oculi . Three patients were cured by tympanotomy with stapedial and tensor tympani tendon section . The aetiology of this type of myoclonus remains unclear . The diagnosis is based on the history of involuntary and rhythmic clicking or buzzing tinnitus which is invariably unilateral . The primary differential diagnosis is palatal myoclonus whilst other local aural pathologies must be excluded by careful clinical assessment . Surgical section of these muscles via tympanotomy brings guaranteed relief when conservative measures fail.

J Pediatr Orthop, 1994 May-Jun, 14(3), 299 - 303
Management of spasticity in cerebral palsy with botulinum-A toxin: report of a preliminary, randomized, double-blind trial; Koman LA et al.; In order to evaluate further the efficacy of local intramuscular injections of botulinum-A toxin (BAT-A) in the management of dynamic equinus deformity associated with cerebral palsy, a randomized, double-blind, placebo-controlled study was undertaken . When evaluated using our Physician Rating Scale, 83% (five of six) of patients receiving toxin showed improvement, versus 33% (two of six) receiving placebo . There were no major complications . BAT-A injections appear to be safe and effective in children, and merit further prospective study.

J Cell Biol, 1994 May, 125(4), 893 - 902
Role of muscle insulin-like growth factors in nerve sprouting: suppression of terminal sprouting in paralyzed muscle by IGF-binding protein 4; Caroni P et al.; The protracted absence of muscle activation initiates complex cellular and molecular reactions aimed at restoring functional neuromuscular transmission and preventing degenerative processes . A central aspect of these reactions is the sprouting of intramuscular nerves in the vicinity of inactivated muscle fibers . Sprouts emerging from terminal nerve branches and nodes of Ranvier can reestablish functional contacts with inactive muscle fibers, and this is an essential restorative process in pathological conditions of the neuromuscular system . Due to their rapid upregulation in inactive skeletal muscle fibers and their ability to induce nerve sprouting in adult muscle, insulin-like growth factors (IGFs) are candidate signaling molecules to promote restorative reactions in the neuromuscular system . In this study we have exploited the high affinity and specificity of IGF-binding protein 4 (IGF-BP4) and IGF-BP5 for IGF1 and IGF2 to determine whether these growth factors are involved in the nerve sprouting reaction in paralyzed skeletal muscle . In tissue culture experiments with sensory- and motoneurons we demonstrate that the neurite promoting activity of IGF1 is blocked by IGF-BP4, and that a similar IGF-BP-sensitive activity is detected in muscle extracts from paralyzed, but not from control muscle . In in vivo experiments, we show that local delivery of IGF-BP4 to Botulinum toxin A-paralyzed skeletal muscle effectively prevents nerve sprouting in that muscle . Our findings indicate that muscle IGFs play an essential role in intramuscular nerve sprouting . In addition, these findings suggest that IGFs are major signaling factors from inactivated muscle to promote local restorative reactions, including interstitial cell proliferation and nerve sprouting.

J Neurosci, 1994 May, 14(5 Pt 2), 3378 - 88
Signaling by insulin-like growth factors in paralyzed skeletal muscle: rapid induction of IGF1 expression in muscle fibers and prevention of interstitial cell proliferation by IGF-BP5 and IGF-BP4; Caroni P et al.; In the absence of muscle activity, muscle fibers, muscle interstitial cells, and intramuscular nerves display characteristic reactions presumably aimed at restoring a functioning neuromuscular system and avoiding degenerative events . In partially denervated muscle these include proliferation of interstitial cells, followed by nerve sprouting . The same reactions can be induced in intact muscle by elevation of intramuscular insulin-like growth factor (IGF) levels . To determine whether IGFs may participate in the initiation of restorative reactions in inactivated muscle we analyzed the expression of IGF1 and IGF2 mRNA in botulinum toxin-paralyzed and in denervated rat skeletal muscle, and interfered with the activity of IGFs by locally applying the IGF-binding proteins IGF-BP5 and IGF-BP4 . To obtain a resolution of the in situ hybridization signals at the cellular level, a nonradioactive method based on digoxigenin-labeled probes was applied . We found that muscle fiber IGF1 mRNA increased rapidly and transiently in inactivated muscle . The time course of this response was similar to that reported for ACh receptor subunits and myogenins . A similar behavior was observed for the corresponding IGF1 protein . The IGF1 response coincided with the transient muscle interstitial cell proliferation reaction . Local application of recombinant IGF-BP5 or IGF-BP4, which are endogenous and specific extracellular ligands of IGFs, prevented the stimulation of interstitial cell proliferation in paralyzed muscle . Our data demonstrate the elevated production of the growth factor IGF1 among early reactions in electrically inactive skeletal muscle fibers and indicate that muscle IGFs are required for the induction of intramuscular interstitial cell proliferation . These findings support the hypothesis that in the neuromuscular system IGF1 plays an important role in initiating cellular reactions in the vicinity of inactive muscle fibers.

J Biol Chem, 1994 Apr 29, 269(17), 12764 - 72
Cleavage of members of the synaptobrevin/VAMP family by types D and F botulinal neurotoxins and tetanus toxin; Yamasaki S et al.; Tetanus toxin (TeTx) and the various forms of botulinal neurotoxins (BoNT/A to BoNT/G) potently inhibit neurotransmission by means of their L chains which selectively proteolyze synaptic proteins such as synaptobrevin (TeTx, BoNT/B, BoNT/F), SNAP-25 (BoNT/A), and syntaxin (BoNT/C1) . Here we show that BoNT/D cleaves rat synaptobrevin 1 and 2 in toxified synaptosomes and in isolated vesicles . In contrast, synaptobrevin 1, as generated by in vitro translation, is only a poor substrate for BoNT/D, whereas this species is cleaved by BoNT/F with similar potency . Cleavage by BoNT/D occurs at the peptide bond Lys59-Leu60 which is adjacent to the BoNT/F cleavage site (Gln58-Lys59) and again differs from the site hydrolyzed by TeTx and BoNT/B (Gln76-Phe77) . Cellubrevin, a recently discovered isoform expressed outside the nervous system, is efficiently cleaved by all three toxins examined . For further characterization of the substrate requirements of BoNT/D, we tested amino- and carboxyl-terminal deletion mutants of synaptobrevin 2 as well as synthetic peptides . Shorter peptides containing up to 15 amino acids on either side of the cleavage site were not cleaved, and a peptide extending from Arg47 to Thr116 was a poor substrate for all three toxins tested . However, cleavability was restored when the peptide is further extended at the NH2 terminus (Thr27-Thr116) demonstrating that NH2 terminally located sequences of synaptobrevin which are distal from the respective cleavage sites are required for proteolysis . To further examine the isoform specificity, several mutants of rat synaptobrevin 2 were generated in which individual amino acids were replaced with those found in rat synaptobrevin 1 . We show that a Met46 to Ile46 substitution drastically diminishes cleavability by BoNT/D and that the presence of Val76 instead of Gln76 dictates the reduced cleavability of synaptobrevin isoforms by TeTx.

Biochem Biophys Res Commun, 1994 Apr 29, 200(2), 829 - 35
Botulinum neurotoxin type G proteolyses the Ala81-Ala82 bond of rat synaptobrevin 2; Yamasaki S et al.; Tetanus toxin and the botulinum neurotoxins types A to F inhibit neurotransmitter release from presynaptic nerve endings by selectively proteolysing the synaptic proteins synaptobrevin, syntaxin, or SNAP-25 . Here, we show that botulinum toxin type G cleaves rat synaptobrevin 2 between Ala81 and Ala82, a peptide bond that differs from those attacked by tetanus toxin and the botulinal toxins types B, D, and F . Synaptobrevin isoforms carrying a Gly in the P1 position are poor substrates . Analyses of N-terminal deletion mutants of rat synaptobrevin 2 showed that a substrate starting at Leu54 is cleaved efficiently, whereas substrates beginning at Leu60 or Phe77 are cleaved partially or not at all, respectively.

Proc Natl Acad Sci U S A, 1994 Apr 26, 91(9), 3564 - 8
Triterpene derivatives that block entry of human immunodeficiency virus type 1 into cells; Mayaux JF et al.; A series of triterpene compounds characterized by a stringent structure-activity relationship were identified as potent and selective inhibitors of human immunodeficiency virus type 1 (HIV-1) replication . Currently studied botulinic derivatives have 50% inhibitory concentrations (IC50) against HIV-1 strain IIIB/LAI in the 10 nM range in several cellular infection assays but are inactive against HIV-2 . These compounds did not significantly inhibit the in vitro activities of several purified HIV-1 enzymes . Rather, they appeared to block virus infection at a postbinding, envelope-dependent step involved in the fusion of the virus to the cell membrane.

J Pharmacol Exp Ther, 1994 Apr, 269(1), 256 - 62
Inhibition of vacuolar adenosine triphosphatase antagonizes the effects of clostridial neurotoxins but not phospholipase A2 neurotoxins; Simpson LL et al.; Bafilomycin A1, an inhibitor of vacuolar adenosine triphosphatase, was tested for its ability to antagonize botulinum neurotoxins (serotypes A-G), tetanus toxin and phospholipase A2 neurotoxins (notexin, beta-bungarotoxin, taipoxin and textilotoxin) on the mouse phrenic nerve-hemidiaphragm preparation . Bafilomycin itself produced concentration-dependent blockade of neuromuscular transmission without blocking nerve action potentials or muscle action potentials . This effect may have been due to inhibition of the proton pump that regulates acetylcholine transport into vesicles . At submaximal concentrations, bafilomycin was very effective in delaying the onset of paralysis due to all clostridial neurotoxins, but it had no protective effect against phospholipase A2 neurotoxins . Experiments were done to determine which of the three steps in clostridial neurotoxin action was antagonized by bafilomycin (e.g., binding, internalization and intracellular poisoning) . Both pharmacological experiments and ligand-binding experiments showed that the drug did not block toxin binding to the plasma membrane . Similarly, pharmacological experiments on the time-dependent effects of bafilomycin showed that the drug did not antagonize the intracellular actions of toxins . The data indicated that bafilomycin acted at the intermediate step of internalization . This is in keeping with the facts that: 1) bafilomycin inhibits vacuolar adenosine triphosphatase, which in turn leads to inhibition of acidification in endosomes and 2) clostridial neurotoxins depend upon acidification of endosomes for translocation to the cytosol . The finding that bafilomycin antagonizes tetanus toxin may provide important clues for understanding how this toxin can act locally to produce flaccid paralysis . The finding that bafilomycin is a universal antagonist that protects against all clostridial neurotoxins may have important implications for developing therapeutic drugs.

Nippon Ganka Gakkai Zasshi, 1994 Apr, 98(4), 404 - 7
{Spectral analysis of electromyograms in the strabismus cases--before and after botulinum toxin injection}; Matsubayashi K et al.; An examination was made on the mechanism of botulinum toxin injection into extraocular muscles with the use of the power spectrum . Four cases of esotropia (11-16 years) were treated with 0.5 to 3.25 units of botulinum toxin injected into the medical rectus muscle . After the injection, power spectral components under 300 Hz were decreased . In the case of repeated injections, the spectrum resembled the pattern of slow fiber components . It is assumed that a malfunction of fast fibers rather than slow ones is related to the effects of botulinum toxin on extraocular muscles.

Ophthalmology, 1994 Apr, 101(4), 783 - 7
The use of botulinum toxin for treatment of acquired nystagmus and oscillopsia; Ruben ST et al.; PURPOSE: To evaluate the role of botulinum toxin A (BTA) in treating patients with diminished visual acuity secondary to acquired nystagmus and oscillopsia . METHODS: Twelve patients with acquired nystagmus causing oscillopsia and reduced vision were treated with injection of BTA . Botulinum toxin A was injected directly into the horizontal recti in three patients, and in nine patients retrobulbar BTA was administered . Injections were given at 3- to 4-month intervals and repeated as long as patients noted improvement in their quality of life . RESULTS: Improvement in visual function varied, and not all patients benefited from the procedure . However, 8 of 12 patients demonstrated a measurable improvement in visual acuity . Transient ptosis was the most common side effect . CONCLUSION: Retrobulbar BTA provides a simple and safe alternative in managing a condition for which alternative treatments are typically unsatisfactory.

Dev Biol, 1994 Apr, 162(2), 539 - 48
Electrical activity in the neuromuscular unit can influence the molecular development of motor neurons; Kalb RG et al.; During the first few weeks of postnatal life spinal motor neurons develop electrophysiological, morphological, and molecular features that are characteristic of adult motor neurons . To understand how the acquisition of the mature neuronal phenotype is regulated, we have examined the expression of the motor neuron cell surface proteoglycan recognized by monoclonal antibody Cat-301 in the hamster . Previously we found that Cat-301 immunoreactivity is not present on motor neurons at birth and that by the end of the second postnatal week all motor neurons are Cat-301-positive . Surgical and pharmacological lesion studies have shown that the onset of Cat-301 expression depends upon input from both large-diameter primary afferents and from supraspinal afferents . Once the Cat-301 proteoglycan is expressed on motor neurons, its continued expression is independent of these inputs . These studies suggested that motor neuron maturation depends upon the coordination of several afferent inputs during the first postnatal weeks of life . Our previous studies could not address whether segmental and descending afferents (i) provide a chemical signal (such as a trophic factor) or (ii) confer a pattern of neuronal activity upon motor neurons that then results in the expression of the Cat-301 proteoglycan . The present experiments examine the role of electrical activity in motor neuron maturation . In normal animals, all sciatic motor neurons are Cat-301-positive by Postnatal Day 19 (P19) . Chronic application of the sodium channel blocker, tetrodotoxin (TTX), to the sciatic nerve in neonatal animals reduces the percentage of Cat-301-positive motor neurons found at P21 by one-third . This reduction is not due to a nonspecific inhibition of all protein synthesis, because the expression of two other motor neuron antigens proceeds normally in TTX-treated neonates . Blockade of neuromuscular transmission in neonates by Botulinus toxin A also reduces the percentage of Cat-301-positive motor neurons . Cat-301 expression is not tied simply to neuronal activity, because chronic application of TTX to the sciatic nerve, or Botulinus toxin A to muscles, in the adult does not reduce Cat-301 expression . These findings indicate that electrical activity generated within the neuromuscular unit in early postnatal life can influence the acquisition of mature molecular properties by motor neurons.

Mol Cell Biol, 1994 Apr, 14(4), 2447 - 56
rac p21 is involved in insulin-induced membrane ruffling and rho p21 is involved in hepatocyte growth factor- and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced membrane ruffling in KB cells; Nishiyama T et al.; Insulin and hepatocyte growth factor (HGF) induced morphologically different membrane rufflings in KB cells . Insulin-induced membrane ruffling was inhibited by microinjection of rho GDI, an inhibitory GDP/GTP exchange regulator for both rho p21 and rac p21 small GTP-binding proteins, but not inhibited by microinjection of botulinum exoenzyme C3, known to selectively ADP-ribosylate rho p21 and to impair its function . This rho GDI action was prevented by comicroinjection with guanosine 5'-(3-O-thio)triphosphate (GTP gamma S)-bound rac1 p21 . In contrast, HGF-induced membrane ruffling was inhibited by microinjection of rho GDI or C3 . This rho GDI action was prevented by comicroinjection with GTP gamma S-bound rhoA p21, and this C3 action was prevented by comicroinjection with GTP gamma S-bound rhoAIle-41 p21, which is resistant to C3 . Microinjection of either GTP gamma S-bound rac1 p21 or rhoA p21 alone induced membrane ruffling in the absence of the growth factors . The rac1 p21-induced membrane ruffling was morphologically similar to the insulin-induced kind, whereas rhoA p21-induced ruffling was apparently different from both the insulin- and HGF-induced kinds . Membrane ruffling was also induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C-activating phorbol ester, but not by Ca2+ ionophore or microinjection of a dominant active Ki-ras p21 mutant (Ki-rasVal-12 p21) . The phorbol ester-induced membrane ruffling was morphologically similar to the rhoA p21-induced kind and inhibited by microinjection of rho GDI or C3 . These results indicate that rac p21 and rho GDI are involved in insulin-induced membrane ruffling and that rho p21 and rho GDI are involved in HGF- and phorbol ester-induced membrane rufflings.

Plast Reconstr Surg, 1994 Apr, 93(5), 913 - 8
Aesthetic indications for botulinum toxin injection; Guyuron B et al.; A clinical trial was undertaken to evaluate the effects of commercially available botulinum toxin on 14 hyperactive corrugator muscles, 14 procerus muscles, one case of congenital aplasia of the depressor labii inferioris muscle, and one case of iatrogenic injury to the ramus mandibularis branch of the facial nerve with paralysis of the depressor labii and mentalis muscles . Of the 31 muscles injected, 28 were appropriately paralyzed with the initial injection . The desired results were obtained in the 3 remaining muscles following a second injection . The ability to frown was nullified in all subjects, resulting in the elimination of glabellar lines . Facial symmetry was achieved in both patients with muscle imbalance . The average duration of the paralysis was 8 weeks, with a range of 2 to 16 weeks . However, this period was prolonged in the latter part of the study with an adjustment of the toxin dose . Our results demonstrate that botulinum toxin injected into overactive facial muscles does produce a predictable and reversible paralysis and eliminates or ameliorates deep frown lines . We also illustrate its use in achieving facial symmetry in one patient with congenitally absent depressor labii inferioris and platysma muscles and in another with postrhytidectomy facial nerve paralysis.

J Biol Chem, 1994 Mar 18, 269(11), 8122 - 7
Specific antibodies against the Zn(2+)-binding domain of clostridial neurotoxins restore exocytosis in chromaffin cells treated with tetanus or botulinum A neurotoxin; Bartels F et al.; Although tetanus and botulinum A neurotoxins are ineffective in cultured chromaffin cells, they will inhibit carbachol-induced release of noradrenaline provided they gain access to the cytosol either through artificial pores generated in the plasma membrane or by binding to incorporated exogenous gangliosides . The block of exocytosis persists for weeks followed by a slow recovery of cell function . When specific anti-botulinum A toxin antibodies are introduced into cells through pores after manifestation of the block by botulinum A neurotoxin, restoration of exocytotic function is accelerated and fully reestablished within 4 days . The same time course of restoration is seen with anti-tetanus toxin antibodies in cells poisoned by tetanus toxin . Since the light chains of the toxins are enzymatically active, we have introduced polyclonal and monoclonal anti-light chain antibodies into the cytosol . Of all light chain antibodies tested, only those directed against the peptide homologous to the zinc-binding sequence, which is present in both neurotoxins, restored exocytosis regardless of which toxin caused the block . These results indicate that the zinc-binding domain is directly involved in the interaction of the light chains with their substrates and that the toxins have to be present continuously to maintain the block.

Ann N Y Acad Sci, 1994 Mar 9, 710, 76 - 87
Botulinum toxin in the treatment of neurological disorders; Denislic M et al.; Botulinum toxin therapy is safe and effective in the treatment of different movement disorders, especially focal dystonias . We reviewed botulinum toxin treatment of 97 patients: 36 had blepharospasm, 41 had torticollis, and 20 had diverse movement disorders . Patients with blepharospasm and torticollis improved markedly after botulinum toxin injections . The most common side effect in BS patients was ptosis (44.4%); in TC patients, it was dysphagia (29.3%) . The mean duration of the improvement in both groups was 3.4 months . Very promising results were obtained also in the heterogeneous group including patients with other focal dystonias and cerebral palsy . On the basis of these results, we concluded that BTA injections must now be considered the mainstay of therapy for focal dystonias and other involuntary movement disorders.

Ophthalmic Surg, 1994 Mar, 25(3), 186 - 8
Control of eyelid retraction associated with Graves' disease with botulinum A toxin; Biglan AW; Two patients had satisfactory control of eyelid retraction associated with thyroid orbitopathy with repeated treatment of the levator palpebrae superioris muscle with botulinum A toxin . The effects of the toxin lasted for 3 to 4 months.

Mov Disord, 1994 Mar, 9(2), 233 - 5
Effectiveness of botulinum toxin type A against painful limb myoclonus of spinal cord origin; Polo KB et al.; Botulinum toxin is now an established treatment for blepharospasm, hemifacial spasm, spasmodic torticollis, and spastic dysphonia . We report the effectiveness of botulinum toxin against painful limb myoclonus of spinal cord origin . The patient, a 16-year-old girl with a pulmonary vascular anomaly, Scimitar syndrome, suffered from an acute spinal cord infarct at age 11 . She was left with paralysis of the right leg and bladder dysfunction . Four years after the original insult, she developed "painful cramping" and involuntary movements of the left thigh, which were unresponsive to a wide range of therapeutic trials . The movements were continuous, rhythmic, and confined to the left quadriceps muscles . Electromyographic examination revealed continuous myoclonic discharges . Treatment with botulinum toxin in the left quadriceps muscles resulted in complete cessation of pain and marked reduction in amplitude of the movements, both clinically and electromyographically . This observation indicates the efficacy of botulinum toxin in the treatment of painful spinal myoclonus.

Mov Disord, 1994 Mar, 9(2), 230 - 2
Limb dystonia following electrical injury; Tarsy D et al.; Electrical injuries of the extremities may cause paralysis, muscle atrophy, sensory deficit, causalgia, and reflex sympathetic dystrophy (RSD) . Limb dystonia has rarely been reported following electrical injury to an extremity, although it may result from cerebral hemisphere electrical trauma . Following electrical injury to the upper extremity, three patients developed limb dystonia accompanied by severe pain and sensory symptoms in two patients and features of RSD in one patient . Two patients received botulinum toxin injections without functional benefit . The mechanism of dystonia following peripheral trauma is unknown but may relate to reorganization of central synaptic connections, possibly in the spinal cord.

Mov Disord, 1994 Mar, 9(2), 213 - 7
Development of resistance to botulinum toxin type A in patients with torticollis; Greene P et al.; Between 1984 and 1992, 559 patients with torticollis were treated with botulinum toxin type A (btx) injections . Twenty-four of these 559 patients (4.3%) had serological evidence of antibodies to btx by mouse neutralization assay . Some of the 559 patients had only one or two injection series, whereas others were lost to follow-up, so that the actual prevalence of serologically detectable antibodies may be higher than 4% . In addition, some patients who improved after btx injections lost benefit and stopped developing muscle atrophy from adequate doses of btx, without serological evidence of antibodies . To evaluate the risk factors for btx resistance (loss of benefit and muscle atrophy after injections with or without serological evidence of antibodies), we reviewed the records of a cohort of torticollis patients injected over 2-45 months (mean, 23 months) beginning in 1988 . Eight of 76 patients (10.5%) developed btx resistance . Compared to nonresistant patients from the same cohort, these eight patients received more frequent injections, had more "booster injections" 2-3 weeks after an initial injection, and received higher doses of btx per treatment . In order to minimize the risk of developing btx resistance, therefore, we recommend that physicians wait as long as possible (at least 1 month) between btx injections, avoid booster injections, and use the smallest possible doses.

J Voice, 1994 Mar, 8(1), 65 - 9
Cellular responses to acute injury: the role of the plasma membrane in cell response to two clostridial toxins; Sherwin JR et al.; The plasma membrane plays an important role in the pathogenesis of acute cell injury . This brief review outlines the role of the plasma membrane in the cellular response to two clostridial toxins, the botulinum C2 toxin and the tetanus toxin . These two toxins belong to the same family of toxins as botulinum toxin type A and type F, those used clinically for the treatment of facial spasm . The actions of C2 toxin on cultured cells give rise to an acute injury characterized by a dissociation of the actin filaments of the cell cytoskeleton . While this toxin can be lethal to intact organisms, the acute cellular response need not necessarily result in cell death . In the case of tetanus toxin, the toxin appears to perturb the plasma membrane so that the function of one important cell second messenger system, protein kinase C, is altered.

Arch Otolaryngol Head Neck Surg, 1994 Mar, 120(3), 310 - 6
Spasmodic dysphonia . Emotional status and botulinum toxin treatment; Murry T et al.; The objectives of this study were to determine the effects of botulinum toxin injection on measures of depression, anxiety, and somatic complaints in patients diagnosed as having spasmodic dysphonia . Patients were asked to complete preinjection questionnaires with self-ratings of depression, state and trait anxiety, and somatic complaints . Approximately 1 week and 2 months following injection, patients were again asked to complete the questionnaires . The spasmodic dysphonic subjects exhibited significantly elevated mean levels of depression and anxiety . These levels were significantly reduced approximately 1 week after injection . Two months later, depression and anxiety measures did not change significantly from their 1-week postinjection values . The results suggest that patients with spasmodic dysphonia who demonstrate significantly elevated measures of depression and anxiety show a reduction in those measures following treatment with botulinum toxin.

Klin Oczna, 1994 Mar, 96(3), 95 - 6
{Treatment of idiopathic blepharospasm with botulinum toxin A}; Toczolowski J et al.; The preparations of botulinum toxin A--Botox and Porton were injected into the orbicular muscle of eye in 18 patients with idiopathic blepharospasm or Meige's syndrome . Improvement was achieved in 17 patients; in 10 injections should be repeated after about 8 weeks . Minor side effects declined after several days . Injection of botulinum toxin could be effective in this disease, otherwise hard to cure.

J Pediatr Ophthalmol Strabismus, 1994 Mar-Apr, 31(2), 85 - 8
Change of eye muscle sarcomeres according to eye position; Scott AB; In three monkeys, the right eye was moved from the primary position to a position of 30 degrees to 45 degrees exotropia by suturing the globe to the orbital wall . In two animals perfused immediately following suturing, histological examination showed the medial rectus and its sarcomeres to be lengthened and the lateral rectus and its sarcomeres to be shortened as compared to those of the unoperated control eye . In the third monkey, after exotropia was maintained for 2 months, the muscle lengths changed but the sarcomere lengths were similar to those of the control eye . We interpret this to indicate addition of sarcomeres to the lengthened medial rectus and removal of sarcomeres from the shortened lateral rectus . This adaptation to conform muscle length to eye position may explain the altered eye position which persists following periods of eye deviation due to muscle paralysis, prior surgery, injury, or botulinum toxin treatment.

J Pediatr Ophthalmol Strabismus, 1994 Mar-Apr, 31(2), 79 - 83; discussion 84
The efficacy of botulinum neurotoxin A for the treatment of complete and partially recovered chronic sixth nerve palsy; Repka MX et al.; Esotropia from chronic sixth nerve palsy or paresis usually requires surgery . Chemodenervation of the antagonist medial rectus muscle, while popular for the treatment of acute sixth nerve palsies and pareses, has not been used extensively for chronic cases . In this study, 22 patients with sixth nerve palsies or partially recovered palsies of greater than 5 months duration were treated with chemodenervation . The etiologies of the sixth nerve palsies were trauma (n = 7), tumor (n = 4), infection/inflammation (n = 3), nerve compression from aneurysm or increased intracranial pressure (n = 4), congenital (n = 1), ischemia (n = 2), and idiopathic (n = 1) . The mean preinjection deviation was 41 prism diopters . A total of 38 injections were administered (mean, 1.7 per patient) . Each patient received an injection of 2.5 to 7.5 units (mean, 4.1) of botulinum neurotoxin A to the ipsilateral medial rectus muscle . Treatment success was assessed 6 months after the last injection . A course of chemodenervation significantly improved the alignment of 9 of the 22 patients (41%) . The mean postinjection deviation was 8 delta . Seven patients (32%) had single binocular vision in primary position restored . These patients had a mean horizontal binocular field of 70 degrees (range, 40 degrees to 100 degrees) . Thirteen patients (59%) had only modest improvement and required surgery . The data suggest that injection of botulinum neurotoxin A is a useful treatment for some patients with chronic sixth nerve weakness . A course of chemodenervation therapy compares less favorably with transposition surgery with concomitant neurotoxin injection for the treatment of these difficult problems.

J Assoc Physicians India, 1994 Mar, 42(3), 205 - 8
Botulinum toxin A in blepharospasm and hemifacial spasm; Behari M et al.; We report the first Indian experience of botulinum toxin A in the treatment of blepharospasm and hemifacial spasm . Sixteen patients, 7 with essential blepharospasm, 5 with Meige syndrome and 4 with hemifacial spasm received botulinum toxin A injection . One patient received 3 courses of injections, 2 received 2 courses and the rest received only one course . The effect was observed after a latent period of less than 48 hours in all patients and lasted for a mean of 16.65 weeks . More than 70% improvement occurred after 17/20 injections (85%) . Poor response was more often seen when blepharospasm was associated with oromandibular dystonia (2/5 injections) . Though the duration of response and subjective score of improvement was best in patients with hemifacial spasm, the numbers were very small for any statistical evaluation . The side effects were local, transient, mild and well tolerated . The commonest side effect was blepharoptosis.

Biochem Mol Biol Int, 1994 Mar, 32(3), 455 - 63
Exogenous zinc ion is required for inhibitory activity of botulinum neurotoxin C1 against norepinephrine release and its endopeptidase activity toward substance P; Yokosawa N et al.; Botulinum neurotoxin C1 inhibited Ca(2+)-evoked norepinephrine secretion from digitonin-permeabilized PC12 cells . The inhibition by the neurotoxin was dependent on the presence of Zn2+ added exogenously . This zinc-dependent inhibition was neutralized by monoclonal antibodies that recognize the sites close to the putative zinc-binding motif in the light chain . The neurotoxin was found to have an endopeptidase activity toward small peptide, substance P . The presence of exogenous Zn2+ was also indispensable to the full expression of this endopeptidase activity . Thus both the inhibition of neurotransmitter release by the C1 neurotoxin and its endopeptidase activity are dependent on exogenous Zn2+, which suggests a strong link between the two activities.

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