J Biol Chem, 1994 Feb 18, 269(7), 5328 - 35 Identification of a vesicle-associated membrane protein (VAMP)-like membrane protein in zymogen granules of the rat exocrine pancreas; Braun JE et al.; Zymogen granules of the exocrine pancreas are the secretory organelles responsible for the regulated secretion of digestive enzymes . Several proteins are associated with or are integral components of the lipid bilayer that forms the zymogen granule membrane . These proteins likely represent important components in the regulated secretion of digestive enzymes . VAMPs (vesicle-associated membrane proteins)/synaptobrevins are a family of 18-kDa integral membrane proteins originally characterized in synaptic vesicles . Polyclonal antisera raised against either a VAMP/glutathione S-transferase (GST) fusion protein or rat brain synaptic vesicles, detected an 18-kDa immunoreactive protein in zymogen granule membranes that co-migrates electrophorectically with rat brain synaptic vesicle VAMP . Rat brain synaptic vesicle VAMP was detected by both antisera . Botulinum-B toxin treatment of zymogen granule membranes did not result in cleavage of zymogen granule membrane VAMP, indicating that exocrine pancreatic VAMP is either VAMP1 or a novel VAMP-isoform . Immunofluorescent studies demonstrated that exocrine pancreatic VAMP localized with GP2, a zymogen granule membrane protein, to the apical region of pancreatic acinar cells . No significant labeling was observed in basolateral regions of pancreatic acinar cells . These results establish the presence of a VAMP protein in the zymogen granule of the rat pancreas and suggest that VAMPs have a role in exocrine secretion. J Immunol, 1994 Feb 1, 152(3), 1370 - 9 Involvement of a botulinum toxin-sensitive 22-kDa G protein in stimulated exocytosis of human neutrophils; Nath J et al.; Studies of human peripheral blood neutrophils (PMNs) demonstrated that botulinum neurotoxin D (BT-D) ADP-ribosylates a 22-kDa PMN G protein (G22k) and inhibits the exocytosis of both specific and azurophilic granules stimulated by FMLP . Furthermore, this inhibition of PMN exocytosis by BT-D was found to be correlated with the degree of irreversible ADP-ribosylation of G22k by BT-D and to require modification of at least 85% of PMN G22k before significant inhibition of secretion is observed . Although both pertussis toxin and BT-D inhibited exocytosis in FMLP-stimulated PMNs, the inhibitory effects of the two toxins were found to be additive . Pertussis toxin and BT-D also inhibited Ca2+/GTP/GTP gamma S-induced secretion in digitonin-permeabilized PMNs, but there were distinct differences between the inhibitory effects of the two toxins . In contrast to BT-D, the exotoxin botulinum C3 was found to ADP-ribosylate primarily a 24- to 25-kDa PMN protein, and it was not found to inhibit Ca(2+)- and GTP-induced secretion in permeabilized PMNs . Ultrastructural studies of BT-D-treated PMNs showed an accumulation of distinct membrane-bound organelles in the periphery of the cells after FMLP stimulation, suggestive of a toxin-induced block in organelle-plasma membrane fusion . Taken together, these findings indicate that BT-D-sensitive G22k has a functional role in stimulated exocytosis of PMNs. Blood Coagul Fibrinolysis, 1994 Feb, 5(1), 63 - 72 Regulation of thrombin-induced endothelial cell activation by bacterial toxins; Patterson CE et al.; It has previously been shown that thrombin effects on endothelial cells can be mediated via G-proteins, which couple the thrombin receptor to several key physiological responses . As G-proteins are known targets of bacterial toxins, specific toxins were used to further characterize G-protein involvement in thrombin activation of bovine pulmonary arterial endothelial cells (BPAEC) and human umbilical vein endothelial cells (HUVEC) . Homogenates were exposed to several bacterial toxins in the presence of 32P-NAD and ADP ribosylation of proteins determined by autoradiography of SDS-PAGE gels . Major substrates were a 40 kDa protein for pertussis toxin, 39, 45 and 52 kDa proteins (Gs) for cholera toxin, a 21 kDa protein for botulinum toxin C, and a 43 kDa protein (actin) for botulinum toxin C2a . The increase in either HUVEC or BPAEC PGI2 release induced by thrombin was not altered by pretreatment with any toxin . However, 1 h treatment of BPAEC monolayers with 1 microgram/ml pertussis toxin resulted in dramatic barrier dysfunction, which was synergistic with the albumin permeability induced by 1 microM thrombin . In contrast, pretreatment with 1 microgram/ml cholera toxin completely prevented the thrombin-induced barrier dysfunction . Moreover, contraction and gap formation due to thrombin challenge, observed by phase contrast microscopy, was greatly augmented by pertussis toxin and prevented by cholera toxin . Whereas 5 micrograms/ml botulinum toxin C did not affect either basal or thrombin-induced barrier dysfunction, botulinum toxin C2a increased basal BPAEC permeability over four-fold . Thus, bacterial toxins have specific and divergent effects on thrombin-induced endothelial cell responses . Botulinum toxin C2a appears to interact directly with actin to produce barrier dysfunction . In contrast, cholera toxin promotes barrier function via its known effects on Gs, stimulating adenylate cyclase and increasing cAMP . Because cholera toxin and pertussis toxin (via inhibition of G(i)) both increase cAMP, yet have opposing effects on barrier function, the present results suggest that pertussis toxin produces barrier dysfunction via ADP ribosylation of a novel G-protein other than G(i) or via a novel action of G(i). Brain, 1994 Feb, 117 ( Pt 1), 27 - 38 Electromyographic features of levator palpebrae superioris and orbicularis oculi muscles in blepharospasm; Aramideh M et al.; Electromyographic (EMG) recording was performed synchronously from the levator palpebrae superioris (LP) and the orbicularis oculi (OO) muscles in 28 patients referred to us for treatment of blepharospasm with botulinum A toxin . At the time of this study, 19 patients were under the treatment with botulinum, four started treatment shortly after the EMG recording and five patients had not yet been treated . Based on the EMG patterns, we were able to classify five major groups of abnormalities . Group 1 (blepharospasm): consisted of 10 patients with dystonic discharges limited to OO, normal LP tonic activity, intact reciprocal inhibition between LP and OO and dense bursts of action potentials with high amplitude preceding the return of LP tonic activity, i.e . 'postinhibition potentiation' of LP, brought about by a brief contraction of OO . Group 2 (combined dystonic activities of LP and OO): seven patients belonged to this group . The EMG recording revealed alternating tremulous discharges in both LP and OO muscles, and short intervals of co-contractions due to moderately disturbed reciprocal inhibition . Group 3 (combination of blepharospasm, LP motor impersistence): the EMG patterns, observed in three patients, were characterized by a gradual cessation of LP activity, followed by a brief contraction of OO, which facilitated the return of LP activity, resulting in opening of the eyes . The EMG recordings, thus, revealed the crucial, beneficial role of postinhibition potentiation as a compensatory mechanism in this type of eyelid movement disorder . The EMG patterns were also characterized by short or prolonged periods of dystonic discharges limited to the OO muscles . Group 4 (combination of blepharospasm, involuntary LP inhibition): this group consisted of four patients . In addition to episodes of dystonic activities of OO, the EMG also showed some periods of involuntary inhibition of LP without any concomitant activities of OO . Two patients also exhibited a failure of inhibition of OO muscle activity, following the voluntary contraction of this muscle . The postinhibition potentiation was often not observed . Group 5 (involuntary LP inhibition): consisted of four patients with EMG patterns of involuntary inhibition of LP activity, without any dystonic discharges in OO . The postinhibition potentiation was not observed in this group . The response to the treatment with botulinum toxin was good in the first group and gradually worsened towards the fifth group . Application of botulinum into multiple sites of OO, especially its pretarsal portion, resulted in better response to the treatment in the second and fourth groups.(ABSTRACT TRUNCATED AT 400 WORDS) Br J Oral Maxillofac Surg, 1994 Feb, 32(1), 29 - 33 Botulinum toxin treatment of bilateral masseteric hypertrophy; Smyth AG; Botulinum toxin is a new and innovative method of treating bilateral masseteric hypertrophy which offers many advantages over conventional surgical treatment . Experience gained through the successful use of this drug when given as an intramuscular injection is reported . No significant side-effects have occurred and this technique is recommended for the routine treatment of masseteric hypertrophy. Br J Oral Maxillofac Surg, 1994 Feb, 32(1), 26 - 8 The medical management of masseteric hypertrophy with botulinum toxin type A; Moore AP et al.; We describe the successful outpatient medical treatment of a patient with bilateral masseteric hypertrophy using botulinum toxin type A in a double-blind placebo controlled study . No significant side-effects occurred, and benefit has so far lasted for 6 months. Ann Neurol, 1994 Feb, 35(2), 237 - 9 Botulinum toxin-A improves the rigidity of progressive supranuclear palsy; Polo KB et al.; Botulinum toxin-A (botox) can improve spasticity and decrease painful spasms in the affected limbs of patients with multiple sclerosis . We report significant improvement of muscle rigidity in the upper limbs after focal administration of botulinum toxin A to 2 patients with progressive supranuclear palsy. Aust N Z J Ophthalmol, 1994 Feb, 22(1), 65 - 7 Retrobulbar botulinum toxin for treatment of oscillopsia; Ruben S et al.; Two cases are presented in which multiple episodic retrobulbar botulinum toxin injections have diminished incapacitating acquired oscillopsia and nystagmus and improved visual acuity . One transient ptosis occurred in six procedures . Improvement duration averaged approximately three months . This is a simple, safe and effective therapy where alternative treatments are typically unsatisfactory. J Biol Chem, 1994 Jan 21, 269(3), 1617 - 20 Proteolysis of SNAP-25 by types E and A botulinal neurotoxins; Binz T et al.; Clostridial neurotoxins, tetanus toxin (TeTx) and the seven related but serologically distinct botulinal neurotoxins (BoNT/A to BoNT/G), are potent inhibitors of synaptic vesicle exocytosis in nerve endings . Recently it was reported that the light chains of clostridial neurotoxins act as zinc-dependent metalloproteases which specifically cleave synaptic target proteins such as synaptobrevin/VAMPs, HPC-1/syntaxin (BoNT/C1), and SNAP-25 (BoNT/A) . We show here that BoNT/E, like BoNT/A, cleaves SNAP-25, as generated by in vitro translation or by expression in Escherichia coli . BoNT/E cleaves the Arg180-Ile181 bond . This site is different from that of BoNT/A, which cleaves SNAP-25 between the amino acid residues Gln197 and Arg198 . These findings further support the view that clostridial neurotoxins have evolved from an ancestral protease recognizing the exocytotic fusion machinery of synaptic vesicles whereby individual toxins target different members of the membrane fusion complex. Science, 1994 Jan 21, 263(5145), 390 - 3 Cell membrane resealing by a vesicular mechanism similar to neurotransmitter release; Steinhardt RA et al.; After injury to the cell membrane, rapid resealing of the membrane occurs with little loss of intracellular contents . This process has been studied by measurement of the rate of dye loss after membrane puncture in both the sea urchin embryo and 3T3 fibroblasts . Resealing of disrupted cell membranes requires external calcium that can be antagonized by magnesium . Block of multifunctional calcium/calmodulin kinase, which regulates exocytotic vesicle availability at synapses, and of kinesin, which is required for outward-directed transport of vesicles, inhibited membrane resealing . Resealing was also inhibited by botulinum neurotoxins B and A, suggesting that the two synaptosomal-associated proteins synaptobrevin and SNAP-25 also participate in resealing . This pattern of inhibition indicates that the calcium-dependent mechanisms for cell membrane resealing may involve vesicle delivery, docking, and fusion, similar to the exocytosis of neurotransmitters. Eur J Biochem, 1994 Jan 15, 219(1-2), 161 - 9 Antagonism of the intracellular action of botulinum neurotoxin type A with monoclonal antibodies that map to light-chain epitopes; Cenci Di Bello I et al.; mAbs were produced in mice against highly purified, renatured light chain (LC) of botulinum neurotoxin A (BoNT A) that was immobilised on nitrocellulose to avoid the undesirable use of toxoids . Subcutaneous implants of relatively high amounts (up to 10 micrograms each) of LC allowed its slow release into the systemic circulation and, thus, yielded much higher antibody titres against the underivatized antigen than had hitherto been obtained by conventional immunization . Seven stable hybridoma cell lines were established which secrete mAb of IgG1 and IgG2b subclasses reactive specifically with BoNT A and LC, in native and denatured states, without showing any cross-reactivity with types B, E, F or tetanus toxin . The pronounced reactivities of three mAbs towards refolded LC or intact toxin, observed in immunobinding and precipitation assays, relative to that seen in Western blots imply a preference for conformational epitopes . Though mAbs 4, 5 and 7 failed to neutralize the lethality of BoNT in vivo, administration intraneurally of mAb7 prevented the inhibition of transmitter release normally induced by subsequent extracellular administration of BoNT A . Notably, the latter mAb reacted with a synthetic peptide corresponding to amino acids 28-53 in the N-terminus of the LC, a highly conserved region in Clostridial neurotoxins reported to be essential for maintaining the tertiary structure of the chain . Most importantly, when mAbs 4 or 7 were microinjected inside ganglionic neurons of Aplysia, each reversed, though transiently, the blockade of acetylcholine release by the toxin; this novel finding is discussed in relation to the nature of the zinc-dependent protease activity of the toxin. Biochim Biophys Acta, 1994 Jan 5, 1199(1), 65 - 8 Involvement of phospholipids in the intoxication mechanism of botulinum neurotoxin; Kamata Y et al.; Phospholipids were examined for their potential to interact with botulinum neurotoxin by an in vivo toxin-inactivation assay and a direct binding assay on a thin layer plate . Type E neurotoxin was inactivated by negatively charged phospholipids, phosphatidylserine (PS) and phosphatidylinositol (PI) . The toxicity of the neurotoxin was not affected by phosphatidylcholine (PC) without an electric charge or phosphatidylethanolamine (PE) with a positive electric charge . The neurotoxin bound directly to PS and PI but not to PC or PE . These results suggest that the negatively charged phospholipids in the cell membranes are involved in the intoxication mechanism of botulinum neurotoxin . The phospholipids PS and PI were tested for their potential to interact within three domains {L, H-1, and H-2} which compose the neurotoxin . All three domains bound to PS; whereas, PI specifically accepted the binding of the H-1 domain relative to the penetration of the neurotoxin into the lipid membrane . In this paper, we discuss the interaction between the neurotoxin and the lipid membrane in the intoxication mechanism. ORL Head Neck Nurs, 1994 Summer, 12(3), 12 - 3 Botulinum toxin therapy for blepharospasm in the otolaryngology clinic; Lassen LF et al.; The use of botulinum toxin (Botox, Allergan, Inc.) is a treatment for spasmodic conditions involving many structures in the head and neck . Proper reconstitution, storage, preparation and administration of Botox are important aspects of its use . This article focuses on the actual preparation and sites of injection of Botox as well as the authors' clinical experiences in using it. Oncogene, 1994 Jan, 9(1), 273 - 9 Involvement of Rho p21 small GTP-binding protein and its regulator in the HGF-induced cell motility; Takaishi K et al.; Hepatocyte growth factor (HGF) induced motility of cultured mouse keratinocytes (308R cells) . This HGF-induced cell motility was inhibited by microinjection of either rho GDI, an inhibitory GDP/GTP exchange protein for rho p21 small GTP-binding protein, or a botulinum exoenzyme C3 which is known to selectively impair the function of rho p21 by ADP-ribosylating its effector domain . The rho GDI action was prevented by comicroinjection with the guanosine 5'-(3-0-thio)triphosphate (GTP gamma S)-bound active form of rhoA p21, and the C3 action was prevented by comicroinjection with a rhoA p21 mutant (rhoAIle41 p21) which is resistant to the C3 action . The HGF-induced cell motility was not inhibited by microinjection of a dominant negative rac1 p21 mutant (rac1Asn17 p21) or a dominant negative Ki-ras p21 mutant (Ki-rasAsn17 p21) . Microinjection of the GTP gamma S-bound form of rac1 p21 or a dominant active Ki-ras p21 mutant (Ki-rasVal12 p21) did not induce cell motility . These results indicate that both rho p21 and rho GDI, but neither rac p21 nor ras p21, are involved in the HGF-induced cell motility . However, microinjection of the GTP gamma S-bound form of rhoA p21 alone did not induce cell motility in the absence of HGF, suggesting that activation of rho p21 is necessary but not sufficient for the HGF-induced cell motility . The HGF-induced cell motility was mimicked by 12-0-tetradecanoyl-phorbol-13-acetate, a protein kinase C-activating phorbol ester, but not by Ca2+ ionophore . The phorbol ester-induced cell motility was also inhibited by microinjection of rho GDI or C3 . These results indicate that both rho p21 and rho GDI are also involved in the phorbol ester-induced cell motility. Laryngoscope, 1994 Jan, 104(1 Pt 1), 8 - 11 Measurement of laryngeal resistance in the evaluation of botulinum toxin injection for treatment of focal laryngeal dystonia; Witsell DL et al.; In the past, there has been no consistent, objective method of following patients undergoing botulinum toxin injections for treatment of laryngeal dystonia . Herein, the application of translaryngeal resistance measurements to 15 dysphonic patients is described . Laryngeal resistance is calculated from analysis of translaryngeal pressure and airflow during the utterance /pi/, and found to fall predictably after successful toxin injection . In our series of patients, laryngeal resistance dropped by 69.1% after initial toxin injection . The changes in resistance over time correlate with subjective impressions of voice quality . Translaryngeal resistance measurements can be used objectively to follow patients longitudinally after injection and to collect objective data for analysis . No previously described measurements have met all these criteria . Laryngeal resistance measurement is an ideal method of documenting the results of botulinum toxin injection for the treatment of focal laryngeal dystonia. Laryngoscope, 1994 Jan, 104(1 Pt 1), 30 - 2 Treatment of adductor laryngeal breathing dystonia with botulinum toxin type A; Grillone GA et al.; Adductor laryngeal breathing dystonia (ALBD) is a rare disorder in which patients have persistent inspiratory stridor, usually normal voice, and cough . Physical exam is characterized by paradoxical movement of the vocal cords on inspiration . These patients have involuntary action-induced spasms of the adductor laryngeal muscles on inspiration . There has been no uniformly satisfactory treatment for the disease . Speech therapy, psychotherapy, and pharmacotherapy have all had limited success . We report the successful use of botulinum toxin type A in seven patients with adductor laryngeal breathing dystonia . All patients received bilateral thyroarytenoid injections . All patients had toxin effect within 72 hours, reaching maximal effect within 2 weeks with sustained improvement for an average of 13.8 weeks . Adverse effects included breathy voice and mild choking on liquids . Both resolved, on average, within 2 weeks . This retrospective study supports the safe and effective use of botulinum toxin type A in the treatment of adductor laryngeal breathing dystonia. Ann Otol Rhinol Laryngol, 1994 Jan, 103(1), 31 - 5 Treatment of dysfunction of the cricopharyngeal muscle with botulinum A toxin: introduction of a new, noninvasive method; Schneider I et al.; Botulinum toxin is known as a relatively safe and efficacious agent for the treatment of various neurologic and ophthalmologic disorders . Since dysphagia and deglutition problems combined with aspiration are often caused by spasticity, hypertonus, or delayed relaxation of the upper esophageal sphincter (UES), conventional treatment including lateral cricopharyngotomy was replaced by localized injections of botulinum toxin into the cricopharyngeal muscle (CM) in a series of 7 patients . The study comprised patients with slight dysphagia caused by isolated hypertonus of the UES, as well as patients with severe deglutition disorders, complete inability to swallow, and aspiration problems . Preoperative diagnostic evaluation included careful history-taking, physical examination, cineradiography, and esophageal manometry to exclude other causes of dysphagia . For precise localization, injections were performed under general anesthesia after location of the CM by direct esophagoscopy and electromyographic guidance . Injections were administered into the dorsomedial part and on both sides into the ventrolateral parts of the muscle . Depending on the severity of symptoms and the intraluminal pressure of the UES, the dose varied between 80 and 120 units (botulinum toxin A from Dysport) . The treatment outcome was evaluated by a disability rating score: patients' complaints were scored by subjective and objective parameters before and after injection . All but 2 patients experienced complete relief or marked improvement of their complaints . There were no severe side effects or postoperative complications . Local botulinum toxin injection proved to be an effective alternative treatment to invasive procedures for patients with isolated dysfunction of the UES, and also for patients with more complex deglutition problems combined with aspiration. Neurology, 1994 Jan, 44(1), 70 - 6 Long-term botulinum toxin treatment of focal hand dystonia; Karp BI et al.; We treated focal hand dystonia in 53 patients with botulinum toxin injections for up to 6 years . Eighty-one percent of the patients improved with at least one injection session . Sixty-five percent of the injections produced transient weakness . We followed 37 of the patients for at least 2 years from the start of treatment, 24 of whom discontinued treatment because of inadequate response, loss of response, inaccessibility of a treatment provider, or the expense of the toxin . Women, who had a greater extent and longer duration of benefit than men, were more likely to continue treatment . The mean interval between injection sessions was 6 months . In most patients, we injected the toxin into the same combination of muscles at each session . The dose of toxin generally fluctuated within a range of 20 units . Side effects were mild and transient and unrelated to the long-term use of botulinum toxin . Botulinum toxin injection is safe and effective for the long-term management of focal hand dystonia. J Ark Med Soc, 1994 Jan, 90(8), 383 - 5 Botulinum toxin in the treatment of adductor spasmodic dysphonia; Thompson AR; Spasmodic dysphonia (SD) is a voice disorder that causes marked disability in the affected individual because of the severe disruption of normal communication that the disorder creates . Of the two distinct types, adductor and abductor SD, the adductor type is the most common and the most amenable to treatment . It is felt to be a neurological problem, but the specific lesion has not been found . The best treatment for adductor SD is injection of botulinum toxin into the thyroarytenoid muscles of the larynx. Biosens Bioelectron, 1994, 9(1), 57 - 63 Detection of botulinum toxin using an evanescent wave immunosensor; Kumar P et al.; Using fluorescein isothiocyanate (FITC)-streptavidin, quartz fibre-immobilized antibody (FiAb) and the evanescent wave component of a light beam, detection of Botulinum Toxin-B (BoTX) is described . Exposure of 3-aminopropyltriethoxysilane/glutaraldehyde (APTS/GA) treated quartz fibres to increasing amounts of anti-BoTX Ab indicated toxin binding to increase in a linear fashion up to approximately 125 ng added Ab . Quantitation of bound BoTX and FiAb by Dot-Blot analysis using avidin-Horseradish peroxidase (HRP) conjugation indicated the presence of 0.27 and 0.67 pmoles, respectively . Inclusion of nonbiotinylated BoTX in sampling mixtures reduced fluorescence in a dose-dependent manner over a narrow concentration range (0-300 ng) . Exposure of FiAb to a variety of venoms resulted in no reduction of BoTX binding suggesting detection of BoTX via immobilized anti-BoTX Ab to be very specific. Klin Monatsbl Augenheilkd, 1994 Jan, 204(1), 10 - 3 {New developments in 1991 in the field of the eyelids}; Sarafiant A et al.; The publications of the year 1991 dealing with new developments in the field of lids have been reviewed . New methods of reconstruction of the upper and lower eye lid are presented . Chondroplast is a substitute for the tarsal plate which is preserved in a new way and is available in an unlimited amount . A four snip procedure for the correction of lower lid entropion is presented . A new technique of ectropion correction which allows great flexibility in the extent of the approximation of the lower lid to the globe is available . New simplified variations of the surgical treatment of Graves' disease provide a predictable height and contour of the eye lid as well as improved cosmetic results . Alternatively to Botulinum toxin Doxorubicin was injected as a treatment for benign essential blepharospasm and hemifacial spasm in an experimental study. J Neuropsychiatry Clin Neurosci, 1994 Winter, 6(1), 50 - 3 Use of botulinum toxin injections for spasmodic torticollis of tardive dystonia; Kaufman DM; Because intramuscular injections of type A botulinum toxin (btx) are effective for idiopathic spasmodic torticollis, they were administered to 3 patients who had neck movements as their only manifestation of tardive dystonia . Each improved, with a decrease in involuntary movement and reduction in pain . None had either systemic or local side effects . Although expensive, btx treatment is recommended for involuntary neck movements of tardive dystonia but not yet for the classic buccolingual dyskinesia. Mov Disord, 1994 Jan, 9(1), 31 - 9 Histologic assessment of dose-related diffusion and muscle fiber response after therapeutic botulinum A toxin injections; Borodic GE et al.; Fiber diameter variability, acetylcholinesterase staining properties, and average fiber diameter were determined 5 weeks after varying doses of botulinum A toxin were administered into albino rabbit longissimus dorsi muscle . The average fiber diameter within the muscle appeared to be a function of the dose of botulinum toxin injected . Fiber diameter variability correlated with the dose of botulinum toxin administered . Both fiber diameter variability and acetylcholinesterase spread characteristics showed a distinct diffusion gradient over a defined field within a muscle . At lower doses (1 IU), collapse of the diffusion gradient occurred over a 15-30-mm segment of muscle . At higher doses (5-10 IU), diffusion of botulinum A toxin effect occurred throughout the entire muscle with no apparent end point . This study demonstrated that botulinum A toxin produces a gradient of denervation in a given muscle and that both the magnitude of denervation and the extent of the gradient are dose dependent . Furthermore, both muscle fiber diameter variability and acetylcholinesterase staining were useful as measures of chemodenervation. Mov Disord, 1994 Jan, 9(1), 104 - 5 Masticatory muscle spasm in a non-Japanese patient with Satoyoshi syndrome successfully treated with botulinum toxin; Merello M et al.; A non-Japanese patient with Satoyoshi syndrome is presented . Severe masticatory muscle spasms interfered with feeding, but were successfully treated with botulinum toxin. Eur J Pharmacol, 1994 Jan 1, 266(1), 19 - 24 Inhibitory effects of botulinum toxin on 5-HT1C receptor-induced Cl- current in Xenopus oocytes; Tohda M et al.; Several low molecular weight G proteins have been identified, but their functional roles remain unclear . To clarify the involvement of low molecular weight G protein in receptor-stimulated turnover of polyphosphoinositide (PI) turnover, influences of botulinum toxins on serotonin (5-HT)-stimulated Cl- current mediated by PI turnover were investigated using Xenopus oocytes injected with rat brain mRNA . Treatment with botulinum toxin C, D or purified ADP-ribosyltransferase of botulinum toxin (botulinum toxin C3 enzyme) inhibited the 5-HT-induced Cl- current in oocytes, and ADP-ribosylated 23 kDa proteins . Both botulinum toxin C3 enzyme-induced inhibition of the current and ADP-ribosylation were suppressed by pretreatment with antibotulinum toxin C3 enzyme antibody . Botulinum toxin D treatment of oocytes was ineffective in the response of Cl- current induced by injection of 50 pmol inositol 1,4,5-trisphosphate and 50 pmol Ca2+ . It is suggested that low molecular weight G proteins ADP-ribosylated by botulinum toxin C3 enzyme are involved in phospholipase C activation in Xenopus oocytes. Microbiol Immunol, 1994, 38(6), 421 - 8 Morphological effects, rate of incorporation, and the enzymatic action of botulinum ADP-ribosyltransferase, known as C3 exoenzyme, on human neuroblastoma GOTO cells; Kamata Y et al.; The susceptibility of various lines of cultured cells to botulinum ADP-ribosyltransferase, known as C3 exoenzyme, was examined . Human neuroblastoma GOTO cells were most sensitive . The C3 exoenzyme caused a change in cell shape that involved extension of neurites . The exoenzyme evoked the outgrowth of neurites from chick ganglion as effectively as nerve growth factor, suggesting that C3 exoenzyme possesses neurotropic activity . Experiments with 125I-labeled enzyme revealed that C3 exoenzyme was rapidly incorporated into cells but the number of incorporated enzyme molecules was small . Once C3 exoenzyme had been incorporated, ADP-ribosylation of the substrate (Rho protein) in GOTO cells occurred immediately and rapidly reached a maximum level . However, some of Rho proteins remained unmodified even after induction of the change in morphology . These findings suggest that ADP-ribosylation by C3 exoenzyme is directly associated with the differentiation of GOTO cells but that other events may also participate in this process. Ter Arkh, 1994, 66(11), 85 - 9 {Bacterial toxins: old functions, a new use}; Malov VA et al.; The paper provides current views on the structure and functional activity of bacterial toxins, clinical use of bacterial lipopolysaccharides and their derivatives as immunostimulators, review investigations on the introduction of the botulinic toxin A in ophthalmological and neurological practice, on production of immunotoxins on the basis of protein-natured bacterial toxins as pharmacologically active therapeutic modalities. Bull Soc Belge Ophtalmol, 1994, 252, 51 - 3 Levator aponeurosis dehiscence in a patient treated with botulinum toxin for blepharospasms and eyelid apraxia; Verhulst S et al.; A 63 years old woman presented with bilateral blepharospasms and eyelid apraxia which was treated with Botulinum toxin injections . A post-treatment ptosis was seen with preserved levator function . It was then suggested that manual eyelid traction could have caused aponeurotic dehiscence of the levator muscles . Surgical exploration confirmed this diagnosis . One must be aware that eyelid apraxia can mask an aponeurotic dehiscence of the levator muscle induced by chronic manual traction exerted on the apractic eyelids. Ann Otolaryngol Chir Cervicofac, 1994, 111(3), 141 - 4 {Treatment of blepharospasm and facial hemispasm with botulinum toxin.Apropos of 58 injections in 22 patients}; Elbaz D; Through this analysis we shall report our experience about botulinum toxin in view of relieving blepharospasm and hemifacial spasm . Many Treatments have been tested with very inconstant often deceptive and sometimes even dangerous results . The present method gives us the possibility to paralyze the spasmed muscles by direct injection into the muscle . After 58 ambulatory injections, 53 effective results were obtained with complete spasm relief for a mean internal of 4 to 5 months . No systemic effect was noticed, but some transient ophthalmic incidents occurred: they can be avoided by simple technical precautions . In hemifacial spasm, the orbicularis eyelid muscle seems to be the "spasm starter" and its unique injection by toxin can often relieve the whole hemiface . This simple method appears remarkably effective, although its duration is limited to a few months; but the injections can be conveniently repeated . However, this technique has to be carefully performed by experienced physicians . It is extremely regrettable to see the enormous price increase of toxin within the last years . This limits a larger use for patients good health. Eye, 1994, 8 ( Pt 5), 511 - 5 Persisting hypotropias following protective ptosis induced by botulinum neurotoxin; Heyworth PL et al.; Botulinum toxin A induced ptosis (BTXAP) has become an established method for producing a temporary ptosis for corneal protection . Adams et al1 reported from an initial series of 15 patients and observed that the ptosis lasted for a mean period of 2.5 weeks and that full recovery was achieved after a mean of 8.1 weeks . They noted that in 80% of cases there was a temporary superior rectus weakness which lasted for a mean of 6 weeks . We present three cases in which the superior rectus weakness was permanent and required corrective strabismus surgery . We believe that these are the first cases reported . We propose two possible mechanisms which may be acting together: firstly that a prolonged period of occlusion may have led to a breakdown of fusion, and secondly that following botulinum toxin induced superior rectus weakness there was contracture of the ipsilateral antagonist muscle further disrupting fusional mechanisms. Doc Ophthalmol, 1994, 86(4), 395 - 402 Orbicular synkinesis after facial paralysis: treatment with botulinum toxin; Roggenkamper P et al.; Involuntary lid closure not rarely accompanies aberrant regeneration of nerve fibers after different types of facial paralysis . 23 patients with such synkinesis were treated with botulinum toxin injections into the orbicularis oculi muscle . After periocular injections all patients showed much improvement: a period of, on the average, 13 symptom-free weeks was followed by a period of minimal symptoms . There were only minor complications . Whenever repeated treatment is necessary, botulinum toxin proves to be an effective therapy for involuntary lid closure after defective healing following facial paralysis. Ann Dermatol Venereol, 1994, 121(10), 721 - 3 {A case of unilateral elastosis with cysts and comedones . Favre-Racouchot syndrome}; Moulin G et al.; INTRODUCTION . Cutaneous elastosis with cysts and comedones (Favre-Racouchot) is one of the oldest known manifestations of helioderma . Both sides of the face are usually involved symmetrically . CASE REPORT . We observed a 65-year-old woman with extremely severe Favre-Racouchot disease localized exclusively on the left side of the face . The diagnosis of elastosis with cysts and comedones was confirmed histologically . This elastosis with cysts and comedones was associated with spasms of the hemiface treated with injections of botulinic toxin . This association was fortuitous and we retained actinic irradiation as the causal agent in this woman who had worked for 15 years in the same room . The elastosis occurred on the side of the face which had been continuously exposed at the same orientation to the window . COMMENTS . This original observation is similar to cases where facial exposure to artificial light or sunlight is asymmetrical, leading to a higher incidence of lesions on one side of the face: colloid milium, actinic keratosis, Dubreuilh melanoma (malignant lentigo) or simple helioderma . The asymmetrical nature of the actinic lesions is often related to automobile driving . This case was particular since it demonstrated that Favre-Racouchot elastosis with cysts and comedones is due to actinic irradiation and not to skin aging. J Fr Ophtalmol, 1994, 17(12), 755 - 68 {Treatment of strabismus with botulinum toxin}; Cordonnier M et al.; Having used botulinum toxin for four years, we describe our experience in one hundred squinting patients and compare our results with the literature . We have good results in unilateral sixth nerve palsy and small deviations with binocular potential . Botulinum toxin can also be helpful in third and fourth nerve palsy, in Graves' ophthalmopathy, as an adjunct to transposition surgery and in cases of under- or overcorrections after surgery . In cases of muscle fibrosis and wide angle strabismus, the results are more disappointing . We describe an original method of conditioning the toxin in individual doses which eases the botulinum consultation processing. Postepy Hig Med Dosw, 1994, 48(5), 505 - 20 {Botulinum toxin: structure, mode of action and therapeutic use}; Tatarkiewicz J; Structure, biological activity, mode and mechanism of action of botulinum toxin as well as its therapeutic use is described . Botulinum toxin type A, one of the most potent biologic toxins, has been found to be of therapeutic value in the treatment of several neurologic and ophthalmologic diseases . Its ability to produce chemical denervation of muscles makes it option for treatment of disorders in which traditional therapeutic procedures are of limited value (e.g . blepharospasm and other focal dystonias, strabismus, spasticity). Bull Soc Belge Ophtalmol, 1994, 254, 85 - 8 Surgical technique for patients with the apractic type of essential blepharospasm: case report; Smet H et al.; A frontalis suspension was carried out in a patient with an essential type of blepharospasm, characterized by difficulties in initiating the act of lid elevation, often referred to as the apractic form of blepharospasm or, as J . Elston proposed, the pretarsal blepharospasm . The patient tries to open the eyes by using the frontalis muscle or by manual traction . It is known than in this form of blepharospasm, insufficient results are seen after botulinum toxin infection . Proper examination of the skin crease of the upper eyelid and of the eyelid gives an idea of the insertion of the levator aponeurosis and of the levator muscle function . A desinsertion, due to frequent manual traction, may be found . In this case, reinsertion of the aponeurosis may relieve the symptoms . If no desinsertion is present a frontalis suspension, similar to those used in ptosis surgery, may give good results. Neurosci Lett, 1993 Dec 24, 164(1-2), 93 - 6 Nerve growth factor induces sensitivity to botulinum neurotoxin type A in norepinephrine-secreting PC12 cells; Banerjee A et al.; Inhibition of Ca(2+)-activated norepinephrine secretion by the botulinum neurotoxin (NT) serotypes A and E was examined in permeabilized PC12 cells . The dichain type E NT reduced with dithiothreitol (DTT) completely inhibited secretion whereas the dichain type A NT reduced with DTT exhibited incomplete inhibitory activity . In contrast, Ca(2+)-activated secretion in PC12 cells treated with nerve growth factor (NGF) was completely inhibited by reduced type A NT . The NGF-treated PC12 cells retained a sensitivity to the type E NT similar to that of untreated PC12 cells . These results indicate that the intracellular mechanisms of inhibition of the types E and A NTs are distinct . NGF appears to either induce the expression of a component selectively required for type A NT sensitivity, or otherwise modifies the secretory apparatus to acquire type A NT sensitivity. Gene, 1993 Dec 22, 136(1-2), 61 - 8 Cloning and characterization of seven novel Dictyostelium discoideum rac-related genes belonging to the rho family of GTPases; Bush J et al.; Cellular processes including proliferation, organization of the actin cytoskeleton, vesicular traffic and secretion of proteins comprising the lysosomal/endosomal system are regulated by low-molecular-weight GTP-binding proteins of the Ras superfamily . However, to date only three Dictyostelium discoideum ras-like genes and two ypt-1/sec4-like genes have been identified and characterized . We report here the identification (using an oligodeoxyribonucleotide probe) of seven additional cDNAs coding for members highly related to the Rac proteins (Ras-related-C3 botulinum toxin substrate) which belong to the Rho (Ras homologous) family of GTPases . Three of these rac-related genes (rac1A, rac1B and rac1C) predict proteins with > 90% amino acid (aa) sequence identity with each other and > 80% identity to the human rac1 gene product, whereas the other members (racA, racB, racC and racD) predict proteins with 46-74% identity to the rac1 and rhoA gene products and to each other . The D . discoideum proteins were entirely conserved over the four regions known to be important for GTP binding and all contained the C-terminal CAAX aa motifs shared by other Rho proteins . Interestingly, the D . discoideum rac-related genes revealed unique patterns of expression during growth and development . For instance, the steady-state level of rac1 mRNA, encoded by three highly related genes, increased transiently during aggregation and then rapidly decreased . In contrast, the cellular abundance of mRNAs encoded by the other rac-like genes decreased at different rates and to different levels during development from the peak levels observed during growth . This suggests that the GTP-binding proteins encoded by these genes may play unique roles during the different stages of the D . discoideum life cycle.(ABSTRACT TRUNCATED AT 250 WORDS) J Comp Neurol, 1993 Dec 22, 338(4), 560 - 74 Modulation of low-affinity nerve growth factor receptor in injured adult rat spinal cord motoneurons; Rende M et al.; Spinal and brainstem motoneurons of the adult rat reexpress low-affinity nerve growth factor receptor (LNGFR) and its mRNA after axotomy . We have previously reported the time courses of this reexpression after cut (no regeneration) or crush (followed by regeneration) of the sciatic nerve . We have shown that the length of the different phases of this reexpression (appearance, maintenance and disappearance) can vary according to the type of axotomy . With the present study we expand our previous data and describe and analyze the modulation the LNGFR expression in adult spinal cord motoneurons following different lesion paradigms . In one approach we have imposed three traumatic injuries that still allow regeneration of the sciatic nerve but with a different time course with respect to the crush injury (application of a silicone regeneration chamber, multiple crushes and delayed repair of ligated nerves) . In a second approach, we have determined the capability of three toxic or metabolic injuries to induce LNGFR expression without any direct trauma of the nerve (experimental diabetogenesis, botulinum and alpha-bungarotoxin intoxication and 2,5-hexanedione intoxication) . In a third approach, we have investigated the effect of the block of the axoplasmic transport on the LNGFR expression following different topical applications of vincristine combined with a nerve crush . The results we present are consistent with the idea that: (1) LNGFR immunoreactivity in adult motoneurons is expressed by motoneurons that are attending to an axonal outgrowth and not a generic signal of cellular damage or impairment of the motor function; (2) LNGFR expression in these motoneurons is related to and parallels the outgrowth process time frame, and (3) the signal/s that trigger and sustain this reexpression may be retrogradely transported from the periphery. J Neurosci Res, 1993 Dec 15, 36(6), 635 - 45 High resolution labeling of cholinergic nerve terminals using a specific fully active biotinylated botulinum neurotoxin type A; Arribas M et al.; We report here on the synthesis and characterization of a fully active biotinylated derivative of the botulinum neurotoxin type A . Different ratios of biotin: botulinum toxin were tested to optimize derivatizing conditions and a ratio of 35:1 was selected for further experiments . The average number of biotin groups per toxin molecule was estimated to be 7.8, occurring at both heavy and light chains, and almost all externally located and easily accessible to recognition by streptavidin . The modified toxin retained its toxicity and its ability to interact with biological membranes . Apart from its suitability for detection in Western blots and in microtiter well plates, biotinylated botulinum toxin proved to be adequate for morphological labeling studies at both light and electron microscopy . Peroxidase histochemistry in cryostat sections of intoxicated rat hemidiaphragm muscles showed a distinct labeling of end-plates . Electron microscopy studies were performed on the electric organ of Torpedo marmorata using colloidal gold-conjugated streptavidin for detection . After intoxication of electric organ fragments with the modified toxin, gold labels were found associated with the presynaptic plasma membrane of nerve terminals and with the membrane of synaptic vesicles . Moreover, the distribution of biotinylated botulinum toxin binding sites over the membrane of synaptosomes isolated from the electric organ of Torpedo and their relationship with intramembrane particles were analyzed using the replica-staining label-fracture technique . It was found that the toxin is never associated with intramembrane particles. Br J Hosp Med, 1993 Dec 15-1994 Jan 18, 50(11), 655 - 9 Consensus statement for the management of focal dystonias; Williams A; Focal dystonias include blepharospasm, torticollis, writer's cramp and laryngeal dystonia . They are not uncommon and are disabling . Botulinum toxin has recently been developed for injection into the muscles in spasm and can relieve symptoms in many patients. Br J Plast Surg, 1993 Dec, 46(8), 703 - 6 Botulinum A chemodenervation: a new modality in cerebral palsied hands; Wall SA et al.; Botulinum A chemodenervation of the Adductor Pollicis muscle for the treatment of the thumb-in-palm deformity in cerebral palsied hands is presented as a new therapeutic option . Early results of a clinical trial in five hemiparetic Cerebral Palsied (C.P.) children are assessed using a prospective nontrialist-biased study design based on an independent panel assessment of pre- and post-intervention photographic and videotaped records of hand function and appearance, in combination with grip dynamometry and goniometry . All cases are shown to improve in terms of both function and appearance with results approaching statistical significance (p = 0.06) when assessed by the Wilcoxon's matched-pairs signed rank test, despite the small study group . The modality is shown to be simple, safe and effective over the period reported (229 days) . The benefit is sustained beyond the period of muscle paresis and ongoing long term follow-up will document the need for, and timing of, reinjection. Am J Phys Med Rehabil, 1993 Dec, 72(6), 364 - 8 Botulinum toxin for the treatment of spasticity in multiple sclerosis . New observations; Borg-Stein J et al.; Potential advantages of intramuscular botulinum toxin for the treatment of spasticity include the lack of sensory effects, ability to target specific muscle groups, ability to weaken muscles in a graded fashion and absence of caustic chemicals such as phenol . We describe the use of botulinum toxin for the treatment of severe lower extremity spasticity in two subjects with multiple sclerosis . Both subjects showed an improvement in spasticity, as measured by the modified Ashworth scale, and in functional status . Both subjects exhibited reductions in muscle tone not only in injected muscles, but also in noninjected muscles in the region . These more distant clinical effects have not been emphasized in previous studies after therapeutic injections of botulinum toxin . Further research is needed to clarify the cause and prevalence of these regional motor effects, as well as to further examine the safety and efficacy of botulinum toxin for spasticity treatment. Ophthalmology, 1993 Dec, 100(12), 1861 - 6 Serum antibody production to botulinum A toxin; Siatkowski RM et al.; PURPOSE: Conflicting data have been reported regarding development of serum antibodies to botulinum A toxin . The purpose of this study is to determine conclusively whether antibody production to this toxin occurs in humans, and, if so, to determine its relationship, if any, to length of treatment, total cumulative dose, and clinical response to treatment . METHODS: Sixty-five sera samples from 42 adults treated with botulinum A toxin for essential blepharospasm, hemifacial spasm, or spasmodic torticollis were analyzed via a sphere-linked immunodiagnostic assay for antibody production . Results were plotted against length of treatment, number of injections, cumulative dose, and treatment effect produced . RESULTS: Twenty-four (57%) of the 42 patients produced antibodies in all three diagnostic groups . No significant differences were found between antibody producers and nonproducers with respect to age (P = 0.216), length of treatment (P = 0.586), number of injections (P = 0.619), or total cumulative dose (P = 0.286) . Within the antibody-producing group, there was no significant correlation between amount of antibody and length of treatment (P = 0.081), number of injections (P = 0.134), or cumulative dose (P = 0.250) . The presence of demonstrable antibodies in serum did not affect the clinical responsiveness to injection . CONCLUSION: Antibody production is present in a majority of patients treated with botulinum A toxin . The sphere-linked immunodiagnostic assay is a reliable and reproducible method for detecting and quantifying these antibodies . When antibody production occurs, it is likely due to variations in individual immune responsiveness and appears to have no direct effect on the patient's clinical response to treatment. Infect Immun, 1993 Dec, 61(12), 5392 - 3 ADP-ribosylation of rainbow trout (Oncorhynchus mykiss) actin by botulinum C2 toxin; Kodama H et al.; Intracellular actin of rainbow trout macrophages was ADP-ribosylated by botulinum C2 toxin, which is composed of two nonlinked protein components, component I and trypsinized component II . The actin in the supernatants of various tissue homogenates of the trout was also directly ADP-ribosylated by component I of C2 toxin, indicating that fish actin other than those of land vertebrates is susceptible to enzymatic modification by component I of C2 toxin. J Clin Neuroophthalmol, 1993 Dec, 13(4), 258 - 61 Botulinum toxin type A in upper lid retraction of Graves' ophthalmopathy; Ebner R; Botulinum toxin type A (BTTA) was injected in the upper lid of 6 patients to reduce palpebral retraction due to Graves' ophthalmopathy . Five unilateral and one bilateral (all female) cases constitute the present series . Injection of 2.5 to 7.5 units of BTTA in the affected lids produced ptosis of 2 to 3 mm in 5 patients . A bilateral case showed a positive but insufficient response by the third injection . An acceptable position of the affected eyelids was maintained for 1 to 8 months . The drug-effect period varied in every patient, regardless of the dose injected, amount of retraction, or endocrine status (hyper, hypo, or euthyroidism) at the moment of treatment . In 5 of 6 patients BTTA provided acceptable upper lid position without cosmetic discomfort . The early results obtained encouraged the use of botulinum toxin in this entity. J Biol Chem, 1993 Nov 25, 268(33), 24535 - 8 ADP-ribosylation of rho p21 inhibits lysophosphatidic acid-induced protein tyrosine phosphorylation and phosphatidylinositol 3-kinase activation in cultured Swiss 3T3 cells; Kumagai N et al.; Botulinum C3 exoenzyme was used to specifically ADP-ribosylate and inactivate rho p21, and the effects of rho p21 inactivation on lysophosphatidic acid (LPA)-induced tyrosine phosphorylation were examined in cultured Swiss 3T3 cells . LPA induced a rapid increase in the tyrosine phosphorylation of a number of proteins . Pretreatment of the cells with the C3 exoenzyme caused ADP-ribosylation of rho p21 in the cells and selectively attenuated the phosphorylation of several proteins, including p43 mitogen-activated protein kinase, p125 focal adhesion kinase, and two proteins of 72 and 88 kDa . C3 exoenzyme pretreatment did not block the initial phosphorylation and activation of mitogen-activated protein kinase but suppressed its subsequent rise . In contrast, the enzyme treatment inhibited the induction of phosphorylation of the 72- and 88-kDa proteins and suppressed the basal and LPA-induced tyrosine phosphorylation of p125 focal adhesion kinase . In addition, immunoprecipitation of cell lysates with an antibody directed against the 85-kDa subunit of phosphatidylinositol 3-kinase (PI 3-kinase) co-precipitated a tyrosine-phosphorylated band of 180 kDa . C3 exoenzyme pretreatment suppressed both the phosphorylation of this band and PI 3-kinase activation associated with LPA stimulation . These findings suggest that rho p21 works as a link between the LPA receptor signal and the subsequent tyrosine phosphorylation and PI 3-kinase activation in these cells. J Biol Chem, 1993 Nov 15, 268(32), 23784 - 7 Identification of the nerve terminal targets of botulinum neurotoxin serotypes A, D, and E; Schiavo G et al.; Botulinum neurotoxins are metalloproteins with one zinc atom bound to the zinc binding motif of zinc endopeptidases . Here we show that botulinum neurotoxin serotypes A, D, and E are zinc endoproteases specific for components of the synaptic vesicle docking and fusion complex . Serotypes A and E cleave SNAP-25, a 25-kDa protein of the synaptic terminal, while serotype D is specific for VAMP/synaptobrevin, a membrane protein of synaptic vesicles . Both rat brain VAMP isoforms are cleaved at a single Lys-Leu peptide bond . The proteolytic activity of these neurotoxins is inhibited by EDTA and captopril. Ger J Ophthalmol, 1993 Nov, 2(6), 426 - 8 Frontalis suspension for essential blepharospasm unresponsive to botulinum toxin therapy . First results; Roggenkamper P et al.; We performed frontalis suspension in 12 patients presenting with essential blepharospasm or "apraxia" of eyelid opening who did not respond sufficiently to botulinum toxin injections . An improvement could be observed in 9 patients . During the follow-up period (4-18 months) the effect of surgical intervention remained stable . As opposed to other surgical procedures (excision of the orbicularis muscle, resection of facial nerve branches), frontalis suspension can be considered as a minimally invasive and even reversible treatment. J Pharmacol Exp Ther, 1993 Nov, 267(2), 720 - 7 Chelation of zinc antagonizes the neuromuscular blocking properties of the seven serotypes of botulinum neurotoxin as well as tetanus toxin; Simpson LL et al.; Botulinum neurotoxin types A, B (unactivated and activated), C, D, E, F and G, as well as tetanus toxin, paralyzed transmission in mouse phrenic nerve-hemidiaphragm preparations . Toxin-induced blockade of transmission was antagonized by chelators {e.g., ethylenediamine tetraacetic acid, tetrakis(2-pyridylmethyl)ethylenediamine or diethylene-triaminepentaacetic anhydride}, but this effect was dependent on incubation conditions . Pretreatment of toxin with chelators failed to produce antagonism, but pretreatment of tissues did produce antagonism . Of the various chelators tested, tetrakis(2-pyridylmethyl)ethylenediamine produced the greatest effect . Antagonism of toxin-induced neuromuscular blockade could be partially reversed by washing chelators from tissues and could be fully reversed by adding an excess of zinc . The ability of chelators to antagonize clostridial neurotoxins was specific and did not extend to phospholipase A2 neurotoxins . Ligand-binding studies with radioiodinated toxin and brain membrane preparations showed that chelators did not antagonize toxicity by inhibiting toxin association with receptors . Similarly, pharmacological experiments with unlabeled toxin- and type-specific antibodies demonstrated that chelators did not act by blocking receptor-mediated internalization of toxin . The chelators appeared to exert their effects by antagonizing the intracellular actions of clostridial neurotoxins . Electrophysiological studies showed that chelators, at concentrations relevant to antagonism of botulinum neurotoxin and tetanus toxin, did not enhance transmitter release.(ABSTRACT TRUNCATED AT 250 WORDS) Eur J Biochem, 1993 Nov 1, 217(3), 965 - 71 Proteolytic cleavage of synthetic fragments of vesicle-associated membrane protein, isoform-2 by botulinum type B neurotoxin; Shone CC et al.; Recent data suggest that botulinum type-B neurotoxin is a protease which acts on vesicle-associated membrane protein, isoform 2 (VAMP-2) . In this report, botulinum type-B neurotoxin is shown to cleave a synthetic fragment (HV62) of VAMP-2, corresponding to the bulk of the hydrophilic domain (amino acids 33-94) . The neurotoxin acts at a single site between Gln76 and Phe77 . Little or no proteolytic activity by botulinum type-B neurotoxin was observed with peptides containing 7, 10 or 20 amino acids spanning the site of cleavage . The proteolytic action of neurotoxin was strongly inhibited by EDTA and o-phenanthroline whereas captopril and phosphoramidon were ineffective . A series of model peptide substrates were synthesised in order to define the smallest VAMP-2 fragment to be cleaved by botulinum type-B neurotoxin . Data obtained from these substrates suggest that the neurotoxin belongs to a novel class of zinc-endoprotease; more than 12 amino acid residues are required on both the NH2- and COOH-terminal side of the cleavage site for optimal proteolytic activity . The results demonstrate that no other components of cellular vesicles are required for the specific action of the neurotoxin on VAMP-2 . The data further show that the highly specific action of the neurotoxin is not dictated solely by the properties of the amino acid residues at the cleavage site but is also dependent on amino acid sequences distal to its site of action. J Neurocytol, 1993 Nov, 22(11), 955 - 65 Differential expression of tenascin after denervation, damage or paralysis of mouse soleus muscle; Irintchev A et al.; The expression of the extracellular matrix molecule tenascin was studied by immunocytochemistry and Western blotting in soleus muscles of adult mice after nerve damage (denervation), muscle injury (induced by enforced running or freezing) and functional block of synaptic transmission (botulinum toxin) . Enhanced expression of tenascin in the extracellular spaces around focally damaged muscle fibres was found already 10 h after onset of running on a motor-driven treadmill which causes muscle injury in soleus muscle . Tenascin expression reached a peak at 2-3 days post-exercise, after which it declined gradually and became undetectable by two weeks after injury . Similarly, cryo-damage of soleus muscles in situ led to upregulation of tenascin . Chronic muscle denervation after sciatic nerve transection caused a persistent (studied up to 31 days) expression of tenascin at denervated endplates and in intramuscular nerve branches but not in other tissue compartments . Local application of botulinum toxin Type A, which results in muscle inactivity but not in tissue degeneration, however, did not induce tenascin expression 12 h to 12 days post-injection . Expression of tenascin after denervation and muscle damage, but its absence after paralysis, were verified by SDS-PAGE and Western blot analysis . Independent of the type of injury (muscle, nerve or both) the known major isoforms of mouse tenascin, as judged by M(r) comparison, were re-expressed, with no preponderance of individual M(r) forms . These results show that tenascin expression in adult muscles is induced by both axon and muscle fibre damage but not by muscle inactivity . In contrast, NCAM, in accordance with previous observations, showed enhanced expression both as a result of inactivity and in association with tissue repair. J Biol Chem, 1993 Oct 5, 268(28), 20838 - 44 A role for the interchain disulfide or its participating thiols in the internalization of botulinum neurotoxin A revealed by a toxin derivative that binds to ecto-acceptors and inhibits transmitter release intracellularly; de Paiva A et al.; Botulinum neurotoxin type A consists of a disulfide-linked light and heavy chain, with an intradisulfide present within the C-terminal half of the latter . The functional consequences of reducing these bonds and alkylating the thiols were investigated . Modification of free cysteine residues had no effect on the toxicity in mouse bioassays or on acetylcholine release in the mouse nerve-diaphragm and the buccal ganglion of Aplysia californica . However, reduction of the toxin prior to alkylation drastically decreased neuroparalytic potency; yet, this derivative inhibited transmitter release if injected directly into a presynaptic neuron in the Aplysia ganglion or added to bovine permeabilized adrenal chromaffin cells . Its antagonism of the action of botulinum neurotoxin A at mammalian motor nerve endings and Aplysia neurons indicates retention of the ability to bind to the toxin's productive ecto-acceptors . Thus, the abolition of the toxicity of extracellularly applied botulinum neurotoxin A by the cleavage of both disulfides, and the alkylation of the half-cystines involved, results from ineffective uptake . Modified forms of the isolated chains of botulinum neurotoxin A were utilized to determine which of the disulfides were necessary for internalization . Alkylation of the cysteines in the light and heavy chains, including those involved in the interchain bond but excluding those of the intact disulfide in the heavy chain, revealed that the intermolecular bond must be present, or the thiols concerned unmodified, for botulinum neurotoxin A to undergo membrane translocation into Aplysia neurons. J Korean Med Sci, 1993 Oct, 8(5), 334 - 40 Botulinum a toxin treatment of hemifacial spasm and blepharospasm; Park YC et al.; We studied the effects of botulinum A toxin in 101 patients with hemifacial spasm and 11 patients with blepharospasm in an open trial and double blind manner . All patients in the open trial and 6 patients in the double blind trial improved after the first injection of botulinum toxin . There was no improvement with placebo . The peak effect ranged from one to 6 days after injection and mean peak effect was 3.6 days in blepharospasm, and 4 days in hemifacial spasm . Of 144 treatments, 98.6% had excellent results, (below grade I) . The duration of beneficial effect ranged 11 to 40 weeks (mean 16.5 weeks) in hemifacial spasm and 9 to 30 weeks (mean 14.2 weeks) in blepharospasm . Complications were encountered in 63.4% in hemifacial spasm and 72.7% in blepharospasm . The common side effects were dry eyes, mouth droop, ptosis and lid edema in order of frequency . These side effects were mild and resolved spontaneously in 1 to 3 weeks . Botulinum A toxin therapy is effective and convenient, and the treatment of choice for patients with hemifacial spasm and blepharospasm. Br J Pharmacol, 1993 Oct, 110(2), 639 - 44 Enhancement by calcitonin gene-related peptide of nicotinic receptor-operated noncontractile Ca2+ mobilization at the mouse neuromuscular junction; Kimura I et al.; 1 . The involvement of calcitonin gene-related peptide (CGRP) in the mechanism of nicotinic acetylcholine receptor-operated noncontractile Ca2+ mobilization (not accompanied by twitch tension) was investigated by measuring Ca(2+)-aequorin luminescence at the neuromuscular junction of mouse diaphragm muscle treated with neostigmine . 2 . Noncontractile Ca2+ transients were enhanced by 4-aminopyridine (100 microM), a K+ channel blocker, and inhibited by botulinum toxin (1-100 micrograms, i.p.) and hexamethonium (10-100 microM), a neuronal nicotinic receptor antagonist . 3 . Noncontractile Ca2+ transients were diminished by CGRP8-37 (10-20 microM), a CGRP antagonist . CGRP (0.3-10 nM) prolonged the duration of noncontractile Ca2+ transients . The effect of CGRP was suppressed by CGRP8-37 (0.1 microM) . 4 . Noncontractile Ca2+ transients were inhibited by H-89 (0.1-1 microM), a protein kinase-A inhibitor . The catalytic subunit of protein kinase-A and AA373 (300 microM), a protein kinase-A activator, prolonged the duration of noncontractile transients . The prolongations either by CGRP or by AA373 were not observed in the presence of H-89 (0.1 microM) . 5 . Contractile (accompanied by twitch tension) but not noncontractile Ca2+ transients were decreased by 12-O-tetradecanoyl phorbol 13-acetate (TPA, 0.3-1 microM), a protein kinase-C activator . Phospholipase A2 increased only contractile Ca2+ transients . Calmodulin-related agents affected neither type of Ca2+ transients . 6 . These results provide the first evidence that nicotinic acetylcholine receptor-operated noncontractile Ca2+ mobilization is promoted by nerve-released CGRP activating protein kinase-A, and is dependent on the accumulated amounts of acetylcholine at the neuromuscular junction where desensitization might readily develop. Mov Disord, 1993 Oct, 8(4), 479 - 83 Use of botulinum toxin type F injections to treat torticollis in patients with immunity to botulinum toxin type A; Greene PE et al.; Fifteen patients with torticollis who had been treated with repeated injections of botulinum toxin type A (botox A) developed antibodies to the toxin . This resulted in loss of benefit in the 13 patients who had improved with botox A injections and failure to develop muscle atrophy after injection in all 15 patients . Patients were then injected with botulinum toxin type F (botox F) in the same muscles that had been injected with botox A . Ten of the 15 improved after botox F injections, including 9 of the 12 patients who had improved with type A toxin . Six of 9 patients with pain had improvement in pain after botox F injections . Patients reported similar improvement with type F and type A toxins, but duration of benefit was approximately 3 months with type A and approximately 1 month with type F . Botox F is an effective treatment for torticollis in patients who are immune to botox A . The usefulness of type F toxin, however, is limited by short duration of benefit. Indian J Ophthalmol, 1993 Oct, 41(3), 121 - 4 Botulinum toxin in the treatment of paralytic strabismus and essential blepharospasm; Thomas R et al.; As an alternative to conventional medical and surgical modalities that have met little success in the treatment of paralytic strabismus and essential blepharospasm, we explored the use of botulinum toxin as a treatment of choice in these two disorders . We used botulinum toxin in three patients with paralytic strabismus and in nine patients with essential blepharospasm . In three patients with paralytic strabismus, the botulinum toxin was injected into the ipsilateral antagonist of the paralysed muscle . The preinjection deviations ranged from 18 to 60 prism diopters . Two of these three patients achieved orthotropia around the thirtieth day and thereafter maintained it . The third patient became orthotropic on the eighteenth day, but deviation recurred and therefore required another injection of toxin . In nine patients with essential blepharospasm, botulinum toxin was injected into the orbicularis oculi muscles . Both objective and subjective improvement occurred in all nine patients within seven days and the effect lasted 12 to 15 weeks . Further injection of the toxin produced extremely beneficial results . However, the only significant complication that we encountered in both groups of strabismus and blepharospasm was ptosis, which was usually partial and temporary . From our experience, we advocate the use of botulinum toxin in the treatment of essential blepharospasm. Neurology, 1993 Sep, 43(9), 1715 - 8 Botulinum antibodies in dystonic patients treated with type A botulinum toxin: frequency and significance; Zuber M et al.; We measured serum antibodies to botulinum toxin (ABT) in 96 patients with focal dystonia who had been treated with type A botulinum toxin . The frequency of detectable ABT was 3% (three patients) . Patients with ABT had received more than 50 ng of botulinum toxin, and the shortest time between two injections was significantly less than in patients without ABT . The clinical evolution of the three patients was heterogeneous: one had decreased effectiveness with repeated injections, another had persistent improvement, and the third never responded to toxin injections. J Neurochem, 1993 Sep, 61(3), 1175 - 8 Antibodies against rat brain vesicle-associated membrane protein (synaptobrevin) prevent inhibition of acetylcholine release by tetanus toxin or botulinum neurotoxin type B; Poulain B et al.; Tetanus and botulinum B neurotoxins are zinc endopeptidases that cleave vesicle-associated membrane protein (VAMP or synaptobrevin) at a single peptide bond . To test the possibility that in vivo also the toxin-induced blockade of neurotransmission is due to cleavage of VAMP, rat brain VAMP-specific antibodies were raised in rabbits . IgGs purified from one antiserum, which bind specifically to rat brain VAMP, also specifically recognize proteins from Aplysia californica in immunoblotting . When injected into neurons in the buccal ganglion of Aplysia, these IgGs did not affect the release of acetylcholine but effectively prevented the inhibitory action of both toxins on neurotransmitter release, thus indicating that the block of neurotransmission by these neurotoxins is consequent to the cleavage of VAMP or specific interaction with VAMP. Arch Otolaryngol Head Neck Surg, 1993 Sep, 119(9), 1018 - 22 Botulinum toxin for the treatment of hyperfunctional lines of the face; Blitzer A et al.; OBJECTIVE: To determine the effectiveness of botulinum toxin injections for the management of hyperfunctional facial lines in patients with dystonia . DESIGN: Twenty-six patients were included in the study: 24 patients had dystonic movement of the face as either a primary or secondary component, and two patients were treated for purely hyperfunctional lines . Botulinum toxin type A was injected via a monopolar hollow-bore Teflon-coated electromyography needle into the facial muscles associated with the hyperfunctional lines . Doses were divided into 1.25- to 10-U aliquots . Qualitative assessments by the patient and physician were made before injection and 2 to 3 weeks after injection . PATIENTS: Twenty-six patients (two male and 24 female) with hyperfunctional lines were included . The ages were from 32 to 84 years with an average age of 59 years . Twenty had dystonia, four had hemifacial spasm, and two had pure hyperfunction without neuromuscular disease . RESULTS: All of the patients had an effect of toxin within the first 24 to 72 hours . All of the patients experienced benefit from the toxin injections with partial or total resolution of painful contractions or unsightly hyperfunctional lines and spasms . The effects of the injection lasted 3 to 6 months . No systemic side effects were noted . Adverse effects included mild, temporary eyelid or lip weakness . CONCLUSION: Based on this initial pilot study, botulinum toxin may be an important new option for the treatment of patients with hyperfunctional facial lines. Muscle Nerve, 1993 Sep, 16(9), 964 - 9 Quantifying how location and dose of botulinum toxin injections affect muscle paralysis; Shaari CM et al.; Despite the widespread use of botulinum toxin to treat muscle dystonias, no method exists to quantify muscle paralysis in either human or nonhuman models . In this study we examined how the location, dose, and volume of botulinum injection affects paralysis in the rat tibialis anterior muscle . Paralysis was quantified by electrically stimulating the nerve to the tibialis anterior and then staining sections of the muscle for glycogen . The areas of glycogen-containing fibers represented regions of botulinum action . The results showed that the most important injection technique is to inject botulinum directly into the motor endplate region of a muscle . Injections only 0.5 cm from the motor endplate resulted in a 50% decrease in paralysis . Increases in dose increased paralysis, however, some of that increase was simply due to the increased volume of injection . Thus, delivering toxin in small volumes near the MEP band of a muscle should produce the most effective paralysis. J Hand Surg {Am}, 1993 Sep, 18(5), 883 - 7 Use of botulinum toxin in the treatment of hand dystonia; Jankovic J et al.; Forty-six patients with hand dystonia, considered disabling despite optimal pharmacologic therapy, were injected in the forearm musculature with botulinum A toxin . Thirty of these patients were followed long enough to provide adequate data for analysis of 86 treatment sessions . There was a 63% female preponderance, with an average age at initial evaluation of 46 years and symptom duration of 7.9 years . Average baseline severity of dystonia was rated as 3.5 on a severity rating scale (0-4 rating; 4 = maximum severity) . The average peak effect response for all injections (79 into wrist flexors and 29 into wrist extensors) was 2.2 for dystonia and 3.0 for pain (0-4 rating; 0 = no response, 4 = maximum benefit) . The latency from injection to onset of effect averaged 5.6 days . Total response duration averaged 9.3 weeks and maximum improvement was 7.5 weeks . Only local complications occurred and consisted primarily of hand weakness (25 patients, 44 sessions) . The results show that botulinum toxin injections effectively control hand dystonia in instances where other forms of therapy have failed. Neurosci Lett, 1993 Aug 20, 158(2), 159 - 62 Ganglioside GD3 enhances adherence of botulinum and tetanus neurotoxins to bovine brain synapsin I; Schengrund CL et al.; Tetanus toxin (TTx) and botulinum toxin serotype A (BTxA), preincubated with trisialoganglioside GT1b, adhere to proteins present on blots of bovine synaptosomal proteins . Differential solubilization and ammonium sulfate fractionation provided material enriched in two proteins that appeared to be adhered to most strongly by the labeled neurotoxins . After excision of the appropriate bands from blots of electrophoretically separated proteins, N-terminal amino acid sequence analysis permitted identification of the proteins as synapsins Ia and Ib . Comparison of the effectiveness of different gangliosides at enhancing adherence of the neurotoxins to blots of synapsins Ia and Ib indicated that GD3 was most effective. Nervenarzt, 1993 Aug, 64(8), 517 - 23 {Local injection treatment with botulinum toxin A in severe arm and leg spasticity}; Konstanzer A et al.; In patients with predominantly focal spasticity, oral antispastic drugs are relatively ineffective or cause unwanted side effects of central origin . Therefore we treated patients disabled by focal spasticity with local injections of Botulinum-Toxin A (Porton Products BOTOX) . Efficacy, dosage, side-effects and injection technique were examined . 11 patients (mean age 48 years) with severe focal spasticity of the flexor muscles of the hand and arm (5 patients), the adductor muscles of the legs (5) or the plantar flexors of the foot (1) due to multiple sclerosis, cervical myelopathy or stroke-related hemi-paresis were treated with BOTOX . Rating scales, including Ashford spasticity scale, pain scale and a hygienic rating scale, were used to evaluate the efficacy . 25 to 30 ng (1000-1200 MU Porton) were injected in the flexor group of the hand or arm and 42 to 50 ng (1680-2000 MU Porton) BOTOX in the adductor group of one leg . 10 of the patients showed an improvement of at least one point on the scales for spasticity, pain and hygiene . Effects could be observed after 4-7 days and lasted for 6-13 weeks . There were no unwanted side-effects . We conclude that BOTOX is an alternative to the systemic application of antispastic drugs . Focal spasticity and pain can be successfully reduced and hygienic care is facilitated. Surg Neurol, 1993 Aug, 40(2), 96 - 103 Selective peripheral denervation for spasmodic torticollis: surgical technique, results, and observations in 260 cases; Bertrand CM; A total of 260 cases of spasmodic torticollis or of the cervical component of diffuse dystonias have been surgically treated with selective peripheral denervation of the involved muscles sparing their antagonists, after verification with electromyography and, if necessary, nerve blocks . Total or marked relief of symptoms with preservation of normal or nearly normal movements has been obtained in 88% of the patients with surgery followed by early physiotherapy . There are minimal sequelae with this approach . Selective denervation may be recommended if, after 2 years, conservative treatment, including botulinum injections, does not offer satisfactory relief of symptoms. J Neurochem, 1993 Aug, 61(2), 501 - 8 Comparison between the effects of botulinum toxin-induced paralysis and denervation on molecular forms of acetylcholinesterase in muscles; Sketelj J et al.; Velocity sedimentation analysis of acetylcholinesterase (AChE) molecular forms in the fast extensor digitorum longus muscle and in the slow soleus muscle of the rat was carried out on days 4, 8, and 14 after induction of muscle paralysis by botulinum toxin type A (BoTx) . The results were compared with those observed after muscle denervation . In addition, the ability of BoTx-paralyzed muscles to resynthesize AChE was studied after irreversible inhibition of the preexistent enzyme by diisopropyl phosphorofluoridate . Major differences were observed between the effects of BoTx treatment and nerve section on AChE in the junctional region of the muscles . A precipitous drop in content of the asymmetric A12 AChE form was observed after denervation, whereas its decrease was much slower and less extensive in the BoTx-paralyzed muscles . Recovery of junctional AChE and of its A12 form after irreversible inhibition of the preexistent AChE in BoTx-paralyzed muscles was nevertheless very slow . It seems that a greater part of the junctional A12 AChE form pertains to a fraction with a very slow turnover that is rapidly degraded after denervation but not after BoTx-produced muscle paralysis . The postdenervation decrease in content of junctional A12 AChE is therefore not primarily due to muscle inactivity . The extrajunctional molecular forms of AChE seem to be regulated mostly by muscle activity because they undergo virtually identical changes both after denervation and BoTx paralysis . The differences observed in this respect between the fast and slow muscles after their inactivation must be intrinsic to muscles. Conn Med, 1993 Aug, 57(8), 565 - 6 Relief of benign essential blepharospasm and ? memory loss by cyproheptadine; Fasanella RM; These results in a small group of patients seem quite clear for the benefit of cyproheptadine in patients with BEB, especially where botulinum A injections fail, are too expensive, or are refused . The apparent memory improvement requires further study . A search for more effective serotonin antagonists without the anticholinergic and antihistaminic side effects of cyproheptadine is needed and is in process. Ophthalmic Surg, 1993 Aug, 24(8), 546 - 50 Acquired blepharoptosis secondary to essential blepharospasm; Bodker FS et al.; We treated four patients with essential blepharospasm, receiving botulinum A toxin, in whom, although they had no preexisting blepharoptosis, a concurrent bilateral acquired blepharoptosis developed . Since the blepharoptosis did not improve after the period of time during which the effects of botulinum A toxin would have been expected to resolve (2 to 10 weeks), we judged that its development was unrelated to the toxin . We propose, rather, that the stretching, attenuation, disinsertion, or dehiscence of the upper eyelid levator muscle caused by the blepharospasm were at least partly responsible for the onset of the blepharoptosis . To ensure appropriate treatment in these cases, careful clinical evaluation is required to differentiate the two conditions. J Neurosci Nurs, 1993 Aug, 25(4), 246 - 8 Oral and facial movements in the aged; Paulson GW et al.; Orofacial movements are often outward manifestations of many generalized organic processes . When muscle weakness and hyperactivity are present, tumors and lesions at the base of the skull should be ruled out . Treatments involving myotomies and denervations are drastic, but can be effective even if disfiguring . Botulinum toxin offers short-term (weeks to months) relief, and when used sparingly the muscles involved are only modestly impaired . Since the etiology of many of these movements is unknown, treatments are usually only palliative and remissions rare. Klin Oczna, 1993 Aug, 95(8), 316 - 9 {Use of botulinum toxin A in treatment of blepharospasm}; Balcewicz I et al.; Retrospective examinations comprised 26 patients with idiopathic, spastic blepharospasm, who received injections of botulinum A toxin subcutaneously or into orbicular muscle of eye . In 6 cases complete recovery was achieved . In all patients, symptoms subsided for a mean period of 12.5 weeks . These results are similar to those described earlier in literature. Trends Microbiol, 1993 Aug, 1(5), 170 - 4 Novel targets and catalytic activities of bacterial protein toxins; Schiavo G et al.; Among bacterial protein toxins with intracellular targets, tetanus and botulinum toxins form a group with unique properties . They are absolutely neurospecific and act in the cytosol of neurons . Recent evidence indicates that they are zinc proteases specific for proteins of the neuroexocytosis apparatus. Ned Tijdschr Geneeskd, 1993 Jul 24, 137(30), 1509 - 12 {Blepharospasm; results of treatment with botulin}; Aramideh M et al.; OBJECTIVE . Discussion of clinical symptoms and differential diagnosis of blepharospasm and treatment with botulinum A toxin . Blepharospasm is an involuntary spasmodic contraction of the eyelids . Within a few years 35%-70% of the patients becomes severely disabled . DESIGN . Prospective, open study . SETTING . Academical Medical Centre, Amsterdam . METHOD . In the period 1985-1992 we have seen 85 patients with blepharospasm . Of these 69 were treated with botulinum toxin, a total of 436 treatments, with a mean dose of 25 IU for each eye . RESULTS . The cause of blepharospasm was unknown in 71 patients . Secondary blepharospasm occurred in: peripheral facial palsy (one patient), herpes zoster infection of the trigeminal nerve (2), brain infarct (1), use of neuroleptics (2), progressive supranuclear palsy (2), Shy-Drager syndrome (1), kernicterus (1), and morbus Sjogren (4) . There were 18 patients with autoimmune diseases . 77 (91%) patients had a (very) severe form of blepharospasm . Electromyographic registration revealed a dysfunction of M . levator palpebrae in 7 patients . More than 70% of the patients were free of symptoms for a mean period of two months after each treatment . Local side effects were seen in 61 (14%) of the 436 treatments: ptosis, haematoma, dry eyes, and diplopia . CONCLUSION . Blepharospasm is a disabling disease and occurs sometimes in association with other neurological and ophthalmological diseases . Botulinum A toxin is a safe and effective therapy . Electrophysiological investigation is important in the differential diagnosis; it is unnecessary to do CT or MRI routinely. J Biomed Eng, 1993 Jul, 15(4), 311 - 8 Mathematical modelling of the enteric nervous network . II: Facilitation and inhibition of the cholinergic transmission; Miftakhov RN et al.; The pharmacokinetic responses of the cholinergic enteric neurone to treatment with acetylcholinesterases, tetrodotoxin, some chloride salts of divalent cations, botulinum toxin, beta-bungarotoxin and changes in the concentration of calcium ions in the external medium and repetitive stimulation are presented . The numerical results obtained reproduce quantitatively the effects of toxins and salts of divalent cations acting at different levels of acetylcholine release from the nerve-terminal . The addition of cholinergic agonists potentiates the action of acetylcholine and increases the amplitude of the generated excitatory postsynaptic potential . A decrease in the concentration of extracellular Ca2+ ions reduces the amplitude of the excitatory postsynaptic potential and significantly increases synaptic transmission time . The effect of tetrodotoxin is the blockade propagation of the action potential along the nerve axon and, as a consequence, acetylcholine release from the vesicular store . All these effects have been shown to be dose-dependent . The repetitive stimulation of the neurone reproduces the effects of accumulation and potentiation . The possible applications of the model for the analysis of the enteric nervous system function are discussed. J Pediatr Orthop, 1993 Jul-Aug, 13(4), 489 - 95 Management of cerebral palsy with botulinum-A toxin: preliminary investigation; Koman LA et al.; Use of intramuscular botulinum-A toxin (Botox) to produce neuromuscular blockade has been effective in treating certain ocular and facial muscular imbalances as well as spasmodic torticollis . In this preliminary open study, the effectiveness of intramuscularly injected Botox on the muscular imbalances of cerebral palsy was assessed in 27 pediatric patients . Each patient had "dynamic deformities" unresponsive to other treatment, and operation was the only other realistic alternative . The dose of Botox was calculated on a unit/body weight basis . In ambulatory patients, clinical changes in gait were assessed by a physician's rating scale . Reduction in spasticity became apparent in 12-72 h after injection; the effect of Botox after target threshold was reached lasted 3-6 months . No major side effects occurred . Botox may prove a useful adjuvant in conservative management of the spasticity of cerebral palsy . Successful management with these injections may allow delay of surgical intervention until the child is older and at less risk of possible complications, including the need for repeated surgical procedures. J Neuroimmunol, 1993 Jul, 46(1-2), 105 - 12 Gangliosides and bacterial toxins in Guillain-Barré syndrome; Willison HJ et al.; Autoimmune factors are strongly favoured as mediating Guillain-Barre syndrome (GBS); however, the precise mechanisms by which this occurs remain unknown . Microbial infections in a susceptible host resulting in an idiosyncratic immune response which cross-reacts with nerve constituents still remains the most plausible working hypothesis on which much current research is based . Considerable recent evidence indicates that this humoral immune response is at least in part directed to gangliosides . Interestingly, many bacterial toxins, including botulinum and tetanus neurotoxins, also bind to gangliosides and induce diseases with some similarities to GBS . This article discusses the evidence in favour of a pathogenic role for anti-ganglioside antibodies in GBS in the context of our knowledge of the biology of gangliosides and the factors that determine their immunogenicity. Neurosurg Clin N Am, 1993 Jul, 4(3), 581 - 91 Management of ophthalmic complications in surgery of the brain stem; Rubenfeld M et al.; This article reviews assessment and rehabilitation of abnormalities of the afferent and efferent visual system that may follow surgery of the brain stem . Attention is drawn to newer techniques of assessment (contrast sensitivities and binocular visual fields) and of management (low vision aids, Fresnel prisms, botulinum toxin injections, and adjustable suture strabismus surgery). Mov Disord, 1993 Jul, 8(3), 371 - 3 Significant improvement of stiff-person syndrome after paraspinal injection of botulinum toxin A; Davis D et al.; Following several months of low back pain, a 36-year-old man developed progressive stiffness of the abdominal, low back, and thigh muscles . On examination, these muscles demonstrated marked hypertonia consistent with the clinical diagnosis of stiff-person syndrome . The patient demonstrated increased lumbar lordosis and had focal hyperhidrosis at different sites . Electromyography showed continuous activity of the paraspinal and thigh muscles, and serum and cerebrospinal fluid antibodies to glutamic acid decarboxylase (GAD) were markedly elevated . Diazepam and Lioresal offered partial pain relief . Paraspinal muscle administration of botulinum toxin A reduced the tone of paraspinal and thigh muscles significantly and resulted in marked improvement of ambulation and cessation of pain. Biochim Biophys Acta, 1993 Jul 1, 1168(3), 299 - 306 Low molecular mass GTP-binding proteins are secreted from mammary epithelial cells in association with lipid globules; Ghosal D et al.; Secretion of milk lipid globules is achieved through encapsulation of triacylglycerol-rich lipid droplets in a specialized region of apical plasma membrane of mammary epithelial cells . A class of low molecular mass GTP-binding proteins were associated tightly with the lipid globule membrane, and these proteins appeared to change from peripheral to integral membrane proteins during intracellular growth and transit of lipid globule precursors . Inclusion of GTP or GTP gamma S in incubation medium stimulated secretion of lipids from primary cultures of permeabilized rat mammary epithelial cells . Six polypeptides with molecular masses between 28 and 21 kDa were detected by ability to bind GTP gamma S following separation of lipid-globule-associated proteins by SDS-PAGE and transblotting onto nitrocellulose . That all of these polypeptides were distinct immunologically from the archetype ras was evident from lack of immunoreactivity with p21 ras G-protein monoclonal antibody in Western blots . This monoclonal antibody bound to a 23 kDa polypeptide of lipid droplets that was not detected with the GTP gamma S binding assay . A 25 kDa component of milk lipid globules was a potent substrate for ADP-ribosylation by botulinum toxin C3, but cholera toxin was much less effective, suggesting that this component may belong to the rac class of G-proteins . The 21 kDa component was related immunologically to ADP ribosylation factor. Biochem Biophys Res Commun, 1993 Jun 30, 193(3), 1311 - 7 Botulinum ADP-ribosyltransferase C3 induces elevation of the vitelline coat of ascidian eggs; Toratani S et al.; Botulinum exoenzyme C3 ADP-ribosylates a 23 kDa protein of unfertilized eggs of the ascidian, Halocynthia roretzi . Microinjection of C3 into the eggs induced elevation of the egg vitelline coat . Co-injection of heparin or EGTA with C3 inhibited the inducing effect of C3 . The vitelline coat of eggs which had been previously co-injected with heparin and C3 was elevated by addition of calcium ionophore, but not by insemination . C3 also induced an increased formation of inositol 1,4,5-triphosphate (IP3) in ascidian egg membranes . Thus the ADP-ribosylation of small GTP-binding protein by C3 seems to be responsible for elevation of the vitelline coat of ascidian eggs through IP3 formation and intracellular calcium mobilization. J Biol Chem, 1993 Jun 5, 268(16), 11516 - 9 Botulinum neurotoxin serotype F is a zinc endopeptidase specific for VAMP/synaptobrevin; Schiavo G et al.; Botulinum neurotoxin serotype F contains the zinc binding motif of zinc endopeptidases . Atomic adsorption analysis of highly purified toxin preparation revealed the presence of one atom of zinc per molecule of toxin, which could be removed with EDTA or o-phenanthroline . The light chain of the neurotoxin was shown to have a zinc-dependent protease activity specific for VAMP/synaptobrevin, an integral membrane protein of synaptic vesicles . Both isoforms of rat VAMP were cleaved at the same site corresponding to the single Gln-Lys peptide bond present in their sequences . This proteolytic activity was inhibited by EDTA, o-phenanthroline, and captopril as well as by VAMP peptides spanning the cleavage site. Presse Med, 1993 Jun 5, 22(20), 957 - 63 {Treatments by local injections of botulinum toxin in neurology . Indications and results}; Soulayrol S et al.; Local injections of botulinum toxin constitute the only truly effective treatment of certain abnormal movements and focal dystonias . The authors describe its indications and report on their personal experience . One hundred and seventeen patients were treated: 48 for blepharospasm, 46 for hemifacial spasm and 23 for spasmodic torticollis . The results were evaluated by means of a score taking into account the effectiveness of treatment, the duration of this effectiveness, the side-effects, if any, observed, and the course of the neurological disorder after several series of injections . The results were good or excellent in 91 percent of patients with hemifacial spasm and 79 percent of patients with blepharospasm . Spasmodic torticollis was much improved in 35 percent of the cases and less, but satisfactorily, improved in 48 percent . In this disease, the muscles which antagonize those responsible for the dystonia must absolutely be re-educated. Clin Neuropharmacol, 1993 Jun, 16(3), 205 - 10 Quantitative electromyographic analysis of changes in muscle activity following botulinum toxin therapy for cervical dystonia; Buchman AS et al.; Reports of efficacy of botulinum toxin for cervical dystonia have relied on subjective reports of improvement or various clinical rating scales . We studied 19 patients with cervical dystonia using Turns analysis to determine if quantitative EMG measures of muscle activity changed following botulinum toxin injections . Before and after botulinum toxin injections, six muscles were evaluated bilaterally . Quantitative EMG analysis of active muscles injected with botulinum toxin showed a significant decline in muscle activity after botulinum toxin (F = {1,41} 55.0; p < 0.001) . Significant reductions in quantitative EMG parameters were also noted in noninjected active muscles after botulinum toxin treatment (F = {1,51} 59.15; p < 0.001) . The sum of EMG activity for all active muscles was calculated for each subject and showed a significant reduction after botulinum toxin injection {MANOVA: (F = {1,5} 5.69; p < 0.05} . Quantitative EMG assessment provides an objective measure of response to botulinum toxin . Decreased muscle activity in noninjected muscles may result from toxin diffusion or reflect relaxation of muscles only secondarily involved in cervical dystonia. Oncogene, 1993 Jun, 8(6), 1449 - 55 ADP-ribosylation of the rhoA gene product by botulinum C3 exoenzyme causes Swiss 3T3 cells to accumulate in the G1 phase of the cell cycle; Yamamoto M et al.; Using botulinum C3 exoenzyme, which specifically ADP-ribosylates the rho gene products (rho proteins), we examined the role of these proteins in cell cycle progression in Swiss 3T3 cells . Incubation of cell lysates with C3 exoenzyme revealed a single {32P}ADP-ribosylated protein with an M(r) of 23K . This protein was identified as rhoA protein by isoelectric focusing and peptide mapping . When C3 exoenzyme was added to the culture, it ADP-ribosylated the substrate protein in the cells and reduced their growth rate and saturation density . The reduction was dependent on the amount of C3 exoenzyme and on the extent of ADP-ribosylation of the rho protein in the cells . Flow cytometric analysis of logarithmically growing cells showed that the enzyme treatment concentration-dependently accumulated the cells in the G1 phase of the cell cycle . When G1-enriched cells were treated with C3 exoenzyme and cell cycle progression initiated by the addition of serum was monitored, inhibition of G1-S transition was clearly observed . These results suggest that the rhoA gene product plays a critical role in G1-S progression in cultured Swiss 3T3 cells and that the ADP-ribosylation abolishes this activity and causes the cells to accumulate in G1 phase. Laryngoscope, 1993 Jun, 103(6), 683 - 92 Botulinum toxin injection for adductor spastic dysphonia: patient self-ratings of voice and phonatory effort after three successive injections; Aronson AE et al.; Ten patients (aged 35 to 70 years) with neurologic adductor spastic dysphonia rated themselves on a 7-point scale of severity for degree of voice improvement and physical effort after a series of three injections of botulinum toxin . Symptoms were noticeably reduced 24 and 48 hours after injection; this improvement was followed by considerable fluctuations in voice quality and phonatory effort . With successive injections, patients differed in their post-injection experiences, the time required to reach optimal voice, and the total duration of benefit . The study shows that the course of voice change after botulinum toxin injection is not predictable, uniform, or equal among patients with spastic dysphonia. J Protein Chem, 1993 Jun, 12(3), 351 - 63 Botulinum type A neurotoxin digested with pepsin yields 132, 97, 72, 45, 42, and 18 kD fragments; Gimenez JA et al.; Botulinum neurotoxin (NT) serotype A is a dichain protein made of a light and a heavy chain linked by at least one interchain disulfide; based on SDS-polyacrylamide gel electrophoresis their molecular masses appear as 147, 52, and 93 kD, respectively . Digestion of the NT with pepsin under controlled pH (4.3 and 6.0), time (1 and 24 hr), and temperature (25 and 30 degrees C) produced 132, 97, 42, and 18 kD fragments . The three larger fragments were isolated by ion-exchange chromatography . The 132 and 97 kD fragments are composed of 52 kD light chain and 72 and 45 kD fragments of the heavy chain, respectively . The sequences of amino terminal residues of these fragments were determined to identify the pepsin cleavage sites in the NT, which based on nucleotide sequence has 1295 amino acid residues (Binz et al., J . Biol . Chem . 265, 9153, 1990) . The 42 kD fragment, beginning with residue 866, is the C-terminal half of the heavy chain . The 18 kD fragment, of which the first 72 residues were identified beginning with residue 1147, represents the C-terminal segment of the heavy chain . The 132 kD fragment (residue 1 to approximately 1146) is thus a truncated version of the NT without its 18 kD C-terminal segment . The 97 kD fragment (residue 1 to approximately 865) is also a truncated NT with its 42 kD C-terminal segment excised . These peptic fragments contain one or two of the three functional domains of the NT (binds receptors, forms channels, and intracellularly inhibits exocytosis of the neurotransmitter) that can be used for structure-function studies of the NT . This report also demonstrates for the first time that of the six Cys residues 453, 790, 966, 1059, 1234, and 1279 located in the heavy chain the later four do not form interchain disulfide links with the light chain; however, Cys 1234 and 1279 contained within the 18 kD fragment form intrachain disulfide . The electrophoretic behaviors of type A NT and its fragments in native gels and their comparison with botulinum NT serotypes B and E as well as tetanus NT suggest that each NT forms dimers or other aggregates and the aggregation does not occur when the 42 kD C-terminal half of the heavy chain is excised . Thus, the C-terminal half of the heavy chain appears important in the self-association to form dimers. J Otolaryngol, 1993 Jun, 22(3), 171 - 5 Unilateral versus bilateral botulinum toxin injections in spasmodic dysphonia: acoustic and perceptual results; Adams SG et al.; The present study compared the effects of unilateral and bilateral thyroarytenoid injections of botulinum toxin (botox) for the treatment of adductor spasmodic dysphonia . Using electromyographic guidance, 15 patients received unilateral botox injections of 15 units each and 11 patients received bilateral injections of 2.5 units in each site . Acoustic recordings of the patient's voice were made prior to injection and at two- and six-week intervals after injection . Both the unilateral and bilateral botox injections were associated with significant improvements in spasmodic dysphonia . This was determined by the acoustic measures of vocal shimmer and the number of voice breaks per second and by the perceptual measures of voice spasm severity . Both types of injections were also associated with a significant increase in vocal breathiness at two weeks post-injection . In addition, a number of acoustic measures including maximum phonation time, vocal jitter, and the number of voice breaks/second indicated that unilateral botox injections may provide both superior and longer lasting benefits than bilateral botox injections. Ital J Neurol Sci, 1993 Jun, 14(5), 361 - 7 Botulinum toxin treatment in patients with focal dystonia and hemifacial spasm . A multicenter study of the Italian Movement Disorder Group; Berardelli A et al.; In six Centers belonging to the Italian Movement Disorder Study Group, the efficacy of botulinum toxin treatment was evaluated in an open collaborative study in 251 patients with focal dystonia and hemifacial spasm . The percentage of functional improvement ranged from 66% to 81% in patients with blepharospasm, from 40% to 51% in patients with spasmodic torticollis and from 73% to 81% in those with hemifacial spasm . Good results were also obtained in patients with oromandibular dystonia, laryngeal dystonia and writer's cramp . Side effects were mild and transient . Local botulinum toxin injection is the first choice symptomatic treatment in focal dystonia and hemifacial spasm. J Voice, 1993 Jun, 7(2), 165 - 71 Perceptual-acoustic relationships in spasmodic dysphonia; Zwirner P et al.; Perceptual ratings were obtained from voice samples of 19 patients with adductor spasmodic dysphonia before and 1 week after unilateral treatment with Botulinum toxin . Five experienced listeners judged samples of sustained phonation using a seven-point equal-interval scale . The perceptual parameters assessed were overall severity, strain-strangled voice quality, and breathiness . Perceptual results were related to the standard deviation of fundamental frequency and the voice break factor, two acoustic parameters previously shown to be significantly improved following Botulinum toxin injection . Results indicate that in general the spasmodic dysphonia voice is perceived as less severe, less strain-strangled, and more breathy 1 week after treatment . Interrelations among perceptual parameters and relationships with acoustic analyses are discussed. Nippon Ganka Gakkai Zasshi, 1993 Jun, 97(6), 757 - 62 {Studies on the botulinum therapy for esotropia improvement of retinal correspondence}; Fukai S et al.; When botulinum therapy was used for esotropia with abnormal retinal correspondence, the esotropia improved effectively and binocular function recovered . Transient overcorrected deviation appeared in 92% of different cases of retinal correspondence . Patients without binocular function and abnormal retinal correspondence of various types were the subjects of this study . There were 18 congenital esotropes and 12 acquired esotropes . Before treatment, there was normal retinal correspondence > abnormal retinal correspondence (NRC > ARC) in 5 cases, abnormal retinal correspondence > abnormal retinal correspondence (ARC > NRC) in 8 cases, abnormal retinal correspondence (ARC) in 16 cases, and lack of retinal correspondence (LRC) in 1 case and the binocular function was incomplete in all cases . Botulinum toxin (0.25-0.5 U) injected into the medial rectus muscles showed hypertonic action . The dose was adjusted according to the deviation angle . We found some changes in retinal correspondence caused by the botulinum therapy in 24 of the 30 patients (80%) . Of these there was NRC in 6, NRC > ARC in 11, and ARC > NRC in 7 . Secondary esotropia was found in 92% of the 24 cases . We believe that improvement of retinal correspondence is due to suppression of anomalous retinal areas with the change of the overcorrected deviation after the usual visual direction disappears . Our results suggest that normal retinal correspondence has a relatively wide range of correspondence area. Masui, 1993 Jun, 42(6), 883 - 7 {Treatment of hemifacial spasm with botulinum toxin}; Tanaka M et al.; We injected botulinum toxin to treat hemifacial spasm, and investigated the effects and the patient's impression of this treatment . Average duration of improvement lasted about 3.5 months in both the initial treatment group and the recurrent group . However the patients in the recurrent group received fewer units of botulinum toxin than those in the initial treatment group . Except for local paralysis that disappeared within a month, there were very few complications . Most patient were satisfied with this treatment . We conclude that the treatment of hemifacial spasm with botulinum toxin is both simple and useful. Axone, 1993 Jun, 14(4), 85 - 8 Local treatment of dystonia and spasticity with injections of botulinum-A toxin; Calne S; The use of botulinum-A toxin will be described in two conditions--the extrapyramidal syndrome of dystonia and the pyramidal deficit, spasticity . There is no cure for dystonia and its cause is unknown . Drug therapy is unpredictable and dose-limiting side effects frequently occur with little or no alleviation of symptoms . Spasticity of adductor muscles in the lower limbs causes profound disability and major nursing problems in patients with chronic disorders of the pyramidal tract . As in the case with dystonia, drug therapy is unsatisfactory . At the UBC Movement Disorders Clinic treatment with botulinum-A has been applied to over 400 patients since 1985 . The results of the first studies using this treatment in spasmodic torticollis (the most common form of focal dystonia) and spasticity (in late stage multiple sclerosis) will be discussed . As well the effects of long term treatment will be addressed . Botulinum-A toxin is approved treatment for strabismus, blepharospasm and hemifacial spasm . Approval for its use in other focal dystonias is anticipated . The very nature of the agent used for treatment requires that patients be well prepared and reassured before they undergo their first treatment . There is a wide gulf between the patients' preconceived notions about the treatment and reality. J Biol Chem, 1993 May 25, 268(15), 11057 - 64 Botulinum toxin inhibits arachidonic acid release associated with acetylcholine release from PC12 cells; Ray P et al.; The molecular mechanisms of depolarization-induced calcium-dependent acetylcholine (ACh) release and its inhibition by botulinum neurotoxin type A (BoTx) are not clear . We studied these mechanisms in an in vitro cholinergic neuronal pheochromocytoma PC12 cell line model . Cultured monolayer PC12 cells were differentiated by treatment with 50 ng/ml nerve growth factor (NGF) for 4 days to enhance cellular ACh synthesis and release . Stimulation of these cells with high K+ (80 mM) in the perfusion medium caused a marked increase (three to four times) in {3H}ACh release in a Ca(2+)-dependent manner . K(+)-stimulated {3H}ACh release was totally inhibited by pretreatment of cells with BoTx (2 nM) for 2 h . High K+ also stimulated the release of arachidonic acid ({3H}AA) from the cell membrane, which was inhibited by BoTx (2 nM) . Addition of phospholipase A2 (PLA2) inhibitors (quinacrine, 4-bromophenacyl bromide, manoalide) to the perfusion medium inhibited K(+)-stimulated {3H}ACh and {3H}AA release in a dose-dependent manner . Inclusion of exogenous AA, the PLA2 activator melittin, or PLA2 itself prevented the effect of BoTx . These results demonstrate that in NGF-differentiated PC12 cells, AA release is associated with ACh release, BoTx inhibits both processes, and increased AA can protect against BoTx. J Hand Surg {Am}, 1993 May, 18(3), 541 - 4 Writer's cramp--a focal dystonia: etiology, diagnosis, and treatment; Rhoad RC et al.; The complaint of hand cramps is common among patients who consult the neurologist or the hand surgeon . Classic writer's cramp is best characterized as a focal dystonia, and electromyographic studies reveal a characteristic pattern of cocontraction of the agonist and antagonist muscles of the forearm and hand . Although the outcome of treatment in the past has been unsatisfying, recent experience with new pharmacologic therapy, such as injections of botulinum toxin, has produced promising results . Further experience and improvement in this area will likely increase the therapeutic success in the treatment of writer's cramp and other focal dystonias. J Neurol Neurosurg Psychiatry, 1993 May, 56(5), 526 - 30 The use of botulinum toxin in the treatment of adductor spasmodic dysphonia; Whurr R et al.; Botulinum toxin injections have been used to treat 31 patients with adductor spasmodic dysphonia . Injections of 3.00-3.75 units of botulinum toxin were performed bilaterally into the thyroarytenoid muscle . This treatment significantly decreased the standard deviation of the fundamental frequency of the speech sample, indicating a reduction in the variability of pitch amongst patients . A total of 96% of patients' subjective diary reports showed an improvement with a median of 7 days to peak effect and a 5 week duration of peak effect. Plast Reconstr Surg, 1993 May, 91(6), 1042 - 5 Botulinum A toxin for treatment of aberrant facial nerve regeneration; Borodic GE et al.; Twelve patients with involuntary synkinetic eyelid closure were given 40 injections of botulinum A toxin . Temporary improvement in involuntary eyelid closure was observed in all 12 patients . Eleven of the 12 patients desired repeated injections . Dose requirements for this indication were compared with doses used in 697 injections in 112 patients with essential blepharospasm and Meige syndrome . Additionally, dose comparisons were made with 269 injections in 71 patients with hemifacial spasm . Dose requirements needed to treat aberrant regeneration of the facial nerve were substantially less than needed to treat blepharospasm and Meige syndrome . The dose requirement was similar to that in hemifacial spasm treatment . The reason for the differences probably relates to existing muscular denervation associated with hemifacial spasm and aberrant facial nerve regeneration. Mayo Clin Proc, 1993 May, 68(5), 443 - 8 Posttraumatic cervical dystonia; Goldman S et al.; Posttraumatic cervical dystonia has been described as a distinct syndrome with some similarities to idiopathic nontraumatic cervical dystonia (torticollis) . We describe five patients in whom cervical dystonia developed immediately after relatively mild trauma to the neck . Four of the five patients had persistent contractions of all cervical muscles including the trapezius muscles, which almost completely prevented motion of the neck and resulted in muscle hypertrophy . The condition persisted unabated in all patients for the entire period of follow-up (duration, 1 1/2 to 3 years) . Pharmaceutical interventions, which had been used previously for idiopathic nontraumatic cervical dystonia, failed to benefit these patients . Two patients who received injections of botulinum toxin had no more than mild benefit . Selective denervation was inapplicable because of the widespread involvement of all cervical muscles in all but one patient . Physical therapy was essentially ineffective . Because of the unusual features and possible medicolegal setting, clinicians may tend to diagnose this condition as a psychogenic disorder or litigation-oriented behavior . The clinical picture, however, is consistent with an organic dystonia that may render the patient functionally and occupationally disabled. Funct Neurol, 1993 May-Jun, 8(3), 193 - 6 Cervical radiculopathy following botulinum toxin therapy for cervical dystonia; Defazio G et al.; The development of transient right-sided lower cervical root impairment is reported in a 49-year-old woman affected by cervical dystonia, who had previously received botulinum toxin injections into the left sternocleidomastoid and the right trapezius . The possibility of a causal relationship was discussed with consideration of: 1) the absence of intercurrent illness or trauma; 2) the positive correlation between sensory-motor disturbances and the botulinum toxin-induced remission of cervical dystonia; 3) the reversibility of cervical radiculopathy (there was no recurrence in the two years following the interruption of the treatment) . Magnetic resonance imaging investigation disclosed an abnormal cervical spine geometry together with moderate spondylosis . We feel that mechanical postural changes following the successful botulinum toxin treatment for cervical dystonia might have played some role in the development of cervical radiculopathy . Thus, skeletal abnormalities should be checked when attempting to correct muscle overactivity by botulinum toxin. Acta Otolaryngol, 1993 May, 113(3), 400 - 4 Botulinum A toxin effects on rat jaw muscle spindles; Filippi GM et al.; Botulinum A toxin (Botox) is used for the treatment of many muscular dystonias . However, the relief of the sustained and abnormal postures induced by Botox administration is not fully explained . In this work the possibility was considered that Botox can produce a block not only at the alpha motor endings, but also at the gamma motor endings, consequently reducing the spindle inflow to the alpha motoneurons, which have a great role in maintaining the tonic myotatic reflex . Jaw muscle spindle discharge was recorded before and after Botox injection in the deep masseter muscle . The drug consistently reduced the spindle afferent discharge . Such an effect is suggested to be direct on gamma endings as: i) muscle tension was not modified by Botox during the recording time; ii) saline administration never changed the spindle discharge . The Botox effect on muscle spindles suggests that the relief from dystonias could be due not only to a partial motor paralysis, but also to a decrease of the reflex muscular tone. J Membr Biol, 1993 May, 134(1), 1 - 13 Triggered exocytosis and endocytosis have different requirements for calcium and nucleotides in permeabilized bovine chromaffin cells; von Grafenstein H et al.; The intracellular requirements for membrane recapture in permeabilized chromaffin cells were compared to the requirements for exocytosis from the same cells . In permeabilized bovine chromaffin cells, calcium-driven exocytosis also triggers, with a short delay, uptake of extracellular horseradish peroxidase (HRP) . This internalized HRP remains compartmentalized within the cell and migrates to a low density band on