Botulism

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 a Percoll gradient which is distinct from the heavier chromaffin granules . The amount of horseradish peroxidase internalized is similar in intact and leaky cells and is approximately equivalent to the volumes secreted . Endocytosis in both preparations is blocked by botulinum toxin, operates in a collapsed membrane potential, and is inhibited by low temperature . In permeabilized cells, exocytosis and coupled endocytosis are activated by the same concentrations of Ca2+ and MgATP . Although secretion requires Ca2+ and MgATP, once exocytosis has occurred the subsequent endocytosis can proceed in the virtual absence of Ca2+ or MgATP, and is largely unaffected by a variety of nucleotide triphosphates (including nonhydrolyzable analogues), and cyclic nucleotides . These data suggest that endocytosis can proceed, once exocytosis has been triggered, under conditions that are quite different from those necessary to support exocytosis, and that the specific requirements for Ca2+ and MgATP in secretion are for the exocytotic limb of the secretory cycle rather than for the associated endocytotic pathway.

Zhonghua Yan Ke Za Zhi, 1993 May, 29(3), 144 - 5
{Treatment of blepharospasm and hemifacial spasm with botulinum A toxin}; Dai Z; 112 patients with blepharospasm or hemifacial spasm were satisfactorily treated by local injections of small doses of botulinum A toxin, resulting in rapid relief or alleviation of the symptoms . No systemic adverse reactions were noted, and local side effects were minor and transient . The study shows that the therapy is safe and effective as a simple and easy alternative of surgical intervention.

J Child Neurol, 1993 Apr, 8(2), 154 - 6
Successful treatment of hereditary trembling chin with botulinum toxin; Gordon K et al.; Hereditary trembling chin is an autosomal dominant condition characterized by recurrent bouts of tremor involving the chin . These episodes are precipitated by emotional upset . There has been considerable debate about the gravity of this condition . This may be a benign movement disorder; however, the rhythmic trembling of mentalis at rest or during times of stress in these patients is often misinterpreted as betraying an incipient emotional upset . For this reason, some patients with this condition may find it socially disabling . We have recently successfully treated one such family with regular botulinum toxin injections to the mentalis muscle.

Mov Disord, 1993 Apr, 8(2), 217 - 9
Postneuroleptic laryngeal dyskinesias: a cause of upper airway obstructive syndrome improved by local injections of botulinum toxin; Feve A et al.; We report a case of laryngeal dyskinesia resulting in severe rest and exercise dyspnea . A 51-year-old man treated for 2 years with flupentixol, an incisive neuroleptic, developed severe dyspnea due to intermittent, rhythmic, and dystonic movements of the vocal cords and upper airway . Local injections of botulinum toxin resulted in spectacular regression of laryngeal spasms and major improvement in breathing . This case emphasizes the risk of upper respiratory dyskinesias associated with neuroleptic treatment and shows the feasibility of a new local treatment in this life-threatening disorder.

Neurology, 1993 Apr, 43(4), 834 - 6
Longitudinal experience with botulinum toxin injections for treatment of blepharospasm and cervical dystonia; Jankovic J et al.; We performed a longitudinal analysis of the effects of repeat botulinum toxin (BTX) injections in 42 patients with blepharospasm (BL) and 115 patients with cervical dystonia (CD) who received BTX injections between 1984 and 1992 in at least five separate treatment sessions . Although the BL patients were significantly older (59.8 years) than the CD patients (50.6 years; p < 0.001), there were no other demographic differences between the two groups . While the total dose per treatment session and the peak effect rating remained the same, the duration of benefit increased and the number of complications decreased during the course of treatment . We concluded that chronic treatment with BTX in patients with BL and CD is not associated with any decline in benefit, and that efficacy may improve slightly with repeat treatments.

Proc Natl Acad Sci U S A, 1993 Apr 1, 90(7), 3048 - 52
Presumptive Renshaw cells contain decreased calbindin during recovery from sciatic nerve lesions; Sanna PP et al.; A subpopulation of calbindin-immunoreactive neurons in lamina VII of the spinal cord has been identified by its location as Renshaw cells, the anatomical substrate for recurrent inhibition . The expression of calbindin (28 kDa) in these calbindin-containing rat ventral horn interneurons was studied with immunocytochemistry after sciatic nerve injuries . One week after axotomy calbindin immunoreactivity was strongly reduced on the lesioned side between levels L4 and L6, while calbindin-containing neurons and fibers were still numerous contralaterally and cranially to the lesioned levels . With the progression of regeneration, calbindin-immunoreactive neurons reappeared, reaching a normal distribution 6-8 weeks after the crush . Similar changes could be mimicked by the intramuscular administration of botulinum toxin . These results suggest that calbindin expression in putative Renshaw cells of the spinal cord might be functionally responsive and that maintenance of calbindin expression may depend on the integrity of motoneurons and neuromuscular transmission.

Baillieres Clin Neurol, 1993 Apr, 2(1), 179 - 85
Use of botulinum toxin in the treatment of cervical dystonia; Poewe W et al.; Idiopathic cervical dystonia, like other adult-onset focal dystonias, has been notoriously difficult to treat . Multiple approaches, including systemic drug treatment with anticholinergics, antidopaminergics, anticonvulsants, muscle relaxants and many other drugs as well as physiotherapy and psychotherapy, usually lead to only temporary amelioration in a minority of patients . Surgical selective EMG-controlled denervation of dystonic neck muscles produces better and, in the hands of some, lasting improvement . However, the procedure is invasive and as such less well accepted by patients . Local injections of botulinum toxin are strikingly successful in improving postural deviation and pain in about 80% of patients . They can be made on an outpatient basis and appear to be safe even over many repeat sessions . Dysphagia is potentially the most serious side-effect but can be decreased in incidence and severity by injecting lower doses, particularly into the sternomastoid . Major drawbacks at present are a lack of prospective data to establish optimal dosage and volume of injection guidelines to preserve good efficacy at a reduced risk of side-effects, and the need to continue indefinitely with repeat injections approximately every 3 months.

Tidsskr Nor Laegeforen, 1993 Mar 10, 113(7), 841 - 3
{Botulinum toxin . A new therapeutic alternative in spastic dysphonia (laryngeal abductor dystonia)}; Loven JO et al.; Laryngeal dystonia is a condition characterized by involuntary spasms of the laryngeal muscles . In most patients this involves the adductor laryngeal muscles (adductor laryngeal dystonia) . Treatment with a variety of therapies, including speech therapy and pharmacotherapy, have led to minimal improvement . Injections of botulinum-toxin (Botox) bilaterally into the vocalis muscles is a new treatment for adductor laryngeal dystonia . Since May 1991 we have treated 23 patients with adductor laryngeal dystonia with botulinum-toxin . In 74% of the patients the voice improved within 24-72 hours, and effect lasted for 2-14 months (average four months) . Most of the patients got a breathy voice and a mild sensation of dysphagia during the first week after the injection.

J Pediatr Ophthalmol Strabismus, 1993 Mar-Apr, 30(2), 88 - 91
Medial rectus electromyographic abnormalities in Duane syndrome; Saad N et al.; We report on four patients from the botulinum toxin clinic at Moorfields Eye Hospital with Duane syndrome, who exhibited paradoxical medial rectus activity . EMGs were performed with a standard toxin needle and were qualitative in nature . Current physiodynamic theories as to the etiology of Duane syndrome are based on an aberrant innervation of the lateral rectus . Paradoxical electromyographic activity of the medial rectus may occur in Duane type I syndrome . A possible explanation for this finding is that the medial rectus may receive aberrant innervation but this may not be physiodynamically significant . We postulate that peripheral innervational anomalies may be secondary to, or coexist with, a brain stem anomaly . If an extraocular muscle possesses a dual innervation, then electromyographic abnormalities, without physiodynamic significance, may occur if the recording electrode comes into contact with muscle fibers innervated by a nerve that supplies a small segment of muscle.

J Clin Neuroophthalmol, 1993 Mar, 13(1), 67 - 71
A comparative study of tear secretion in blepharospasm and hemifacial spasm patients treated with botulinum toxin; Price J et al.; In the neuro-ophthalmology clinic at St . Vincent's Hospital, Melbourne, 57 patients with blepharospasm and 50 patients with hemifacial spasm were treated with botulinum toxin . Schirmer tear tests were conducted on all the patients prior to each treatment and at 1 week following treatment where possible . The results were compared with a control group of 107 patients selected by age and sex . The blepharospasm patients were found to have a significantly lower tear secretion than that of the control group, using the Mann-Whitney test (median = 3.5 mm, compared with median-11.0 mm, p < .0001) . This did not improve following treatment . The patients with hemifacial spasm did not have significantly different tear secretion from that of the control group (t = 1.0, p > .05) . To investigate whether there was any relationship between the symptoms and the result of the Schirmer test, a survey was also conducted on the patients with blepharospasm and hemifacial spasm regarding symptoms, frequency, and type of drops/ointment used.

Neuroscience, 1993 Mar, 53(2), 547 - 52
Comparison of the intracellular effects of clostridial neurotoxins on exocytosis from streptolysin O-permeabilized rat pheochromocytoma (PC 12) and bovine adrenal chromaffin cells; Ahnert-Hilger G et al.; The inhibitory effects of tetanus toxin, botulinum toxin A, their constituent light chains, and botulinum toxin B were compared using streptolysin O-permeabilized rat pheochromocytoma (PC 12) and bovine adrenal chromaffin cells in primary culture . In both types of chromaffin cells exocytosis can be triggered by micromolar amounts of free Ca2+, bovine adrenal chromaffin cells in addition require ATP . In PC 12 cells the isolated tetanus toxin light chain alone blocks exocytosis without any additive . The time-course of the inhibitory action of tetanus toxin light chain in permeabilized PC 12 cells in the absence of ATP is similar to the one obtained with permeabilized bovine adrenal chromaffin cells, in the presence of ATP . Thus, ATP does not seem to be crucial for tetanus toxin (two-chain form) poisoning . Botulinum toxin B (two-chain form), if preactivated by dithiothreitol, also inhibits exocytosis from permeabilized PC 12 cells up to 90% in the absence of ATP . By contrast, botulinum toxin A (two-chain form) or its isolated light chain, which are highly potent in permeabilized bovine adrenal chromaffin cells, causes only a weak inhibition in PC 12 cells . In streptolysin O-permeabilized bovine adrenal chromaffin cells omission of ATP during the incubation with the toxin increases the potency of botulinum toxin A light chain . Under the same conditions the effect of tetanus toxin light chain remains unchanged . Tetanus toxin and botulinum toxin B (two-chain forms) probably block a step which occurs during exocytosis from both PC 12 cells and adrenal chromaffin cells and which could be closely related to the final fusion event.(ABSTRACT TRUNCATED AT 250 WORDS)

Biokhimiia, 1993 Mar, 58(3), 438 - 55
{Signal-conducting and low molecular weight GTP-binding proteins from the lung and endothelium: localization in membranes and cytosol, interaction with F-actin}; Panchenko MP et al.; The following proteins have been identified in mammalian lung and endothelium, using {32P}ADP-ribosylation by bacterial ADP-ribosyltransferase, immuno- and {alpha-32P}GTP-blottings: 41 kDa Gi1 alpha, 40 kDa Gi2 alpha, 41 kDa Gi3 alpha, 40 kDa and 45 kDa subunits of GS alpha, 36 kDa beta 1 and 35 kDa beta 2 subunits of signal-transmitting GTP-binding proteins (G-proteins), the 19-26 kDa low molecular weight GTP-binding proteins (SMG-proteins) ras, rho, rac, G25K (Gp), as well as ARF and SMG proteins binding with a high affinity to {alpha-32P}GTP . These G- and SMG-proteins are contained in various proportions in membrane and cytosol fractions of lung and endothelium cells . Subunits Gi2 alpha and GS alpha (but not beta 1 or SMG-proteins) my partially (approximately 1%) dissociate from the membrane by the action of the GTP analogs GTP{S} or Gpp(NH)p in the presence of magnesium ions . Extraction with low ionic strength buffer solutions in the presence of EDTA is accompanied by the release of G-actin sensitive to whooping cough toxin Gi2 alpha and beta i subunits . The functionally coupled into a alpha beta gamma heterodimer Gi-protein subunits (predominantly Gi2 alpha and beta i) present in the cytosol fraction as well as the SMG-proteins revealed by {alpha-32P}GTP-blotting (but not the SMG-proteins sensitive to the botulinic C3 exoenzyme, rho/rac, or ARF, may interact with F-actin . Approximately 20% of these proteins are associated with the Triton X-100 insoluble (cytoskeletal) fraction of the endothelium . A conclusion is drawn that interactions of G- and SMG-proteins with actin filaments may be the reason for the formation of "multidisperse" structure in a cell.

Neuropharmacology, 1993 Mar, 32(3), 285 - 9
Calcium-dependent release of norepinephrine from permeabilized PC12 cells is inhibited by approximately 48 and approximately 112 kDa fragments of botulinum neurotoxin type E; Lomneth R et al.; Permeabilized PC12 cells exhibit a Ca(2+)-stimulated norepinephrine secretory pathway which is sensitive to botulinum neurotoxin serotypes A, B and E {Lomneth R., Martin T.F.J . and DasGupta B . R . (1991) J . Neurochem . 57: 1413-1421} . Two novel amino terminal fragments of the 150 kDa neurotoxin serotype E (approximately 112 and 48 kDa), produced by digestion with pepsin, were tested in permeabilized PC12 cells . The intracellular inhibitory activity of the approximately 112 kDa amino terminal fragment, like that of the 150 kDa neurotoxin, was progressively enhanced after trypsinization and dithiothreitol reduction . The approximately 50 kDa C-terminal half of the heavy chain therefore does not contribute to the enhancement of inhibitory activity . The approximately 48 kDa amino terminal light chain-like fragment completely inhibited release of norepinephrine, with an IC50 = 500 pM (more potent than the light chain isolated after digestion with trypsin) not requiring reduction with dithiothreitol . These results clarify the molecular basis of activation of neurotoxin by trypsin and dithiothreitol.

Toxicon, 1993 Mar, 31(3), 307 - 17
Effects of stonefish (Synanceia trachynis) venom on murine and frog neuromuscular junctions; Kreger AS et al.; The neuromuscular toxicity of stonefish (Synanceia trachynis) venom was characterized by electrophysiological and electron microscopic examination of isolated murine and frog nerve-skeletal muscle preparations exposed to various concentrations of venom . Low concentrations of venom (2.5-10 micrograms/ml) acted presynaptically by causing release and depletion of neurotransmitter from the nerve terminal . The response was Na+ channel-independent (resistant to tetrodotoxin), required the presence of either Ca2+ or Mg2+, and was observed with botulinum neurotoxin-paralyzed nerve-muscle preparations . Higher concentrations of venom (100-300 micrograms/ml) acted postsynaptically and presynaptically . They caused irreversible depolarization of muscle cells and microscopically observable muscle and nerve damage . We conclude that the previously observed neuromuscular toxicity of stonefish venom is a consequence of the venom's dose-dependent, presynaptic and postsynaptic actions at the myoneural junction.

Ophthalmology, 1993 Mar, 100(3), 318 - 22
Botulinum alignment for congenital esotropia; Ing MR; BACKGROUND: Botulinum toxin injection into the medial rectus has been recommended by several investigators as an alternative to incisional surgery for treatment of patients with congenital (essential infantile) esotropia . Currently, there are no published studies demonstrating both the motor and sensory results of congenital esotropic patients aligned by botulinum toxin . METHODS: The author traveled to two medical centers to personally and objectively examine, with standardized testing methods, 12 patients with congenital esotropia who had been aligned for a minimum of 6 months by the age of 2 years by other investigators . The selected patients had been followed for a minimum of 3 years and were of sufficient maturity to reliably respond to sensory testing . A comparison was made between the author's conclusions about the binocularity results of these patients and the assessment of the treating ophthalmologists . RESULTS: Only 6 of the 12 patients demonstrated optimum motor alignment to within 10 prism diopters (PD) of orthophoria at the time of the study . A minimum of 1-month (average, 5 months) post-botulinum injection was found to be necessary to establish this alignment . Only three of these six aligned patients could both fuse and demonstrate gross stereopsis without the assistance of compensatory prisms . These results can be contrasted to a previously reported group of surgically aligned cases in which 66 of 90 patients aligned by 2 years of age could both fuse and demonstrate stereopsis, without any use of compensatory prisms . CONCLUSION: These results must be considered preliminary . However, alignment by botulinum appears to be less effective in establishing evidence for binocularity than incisional surgery in the treatment of congenital esotropia (P < 0.001).

J Cell Biol, 1993 Mar, 120(5), 1187 - 95
Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI); Kishi K et al.; Evidence is accumulating that the rho family, a member of the ras p21-related small GTP-binding protein superfamily, regulates cell morphology, cell motility, and smooth muscle contraction through the actomyosin system . The actomyosin system is also known to be essential for cytoplasmic division of cells (cytokinesis) . In this study, we examined the action of rho p21, its inhibitory GDP/GTP exchange protein, named rho GDI, its stimulatory GDP/GTP exchange protein, named smg GDS, and botulinum ADP-ribosyltransferase C3, known to selectively ADP-ribosylate rho p21 and to impair its function, in the cytoplasmic division using Xenopus embryos . The sperm-induced cytoplasmic division of Xenopus embryos was not affected by microinjection into the embryos of either smg GDS or the guanosine-5'-(3-O-thio)triphosphate (GTP gamma S)-bound form of rhoA p21, one member of the rho family, but completely inhibited by microinjection of rho GDI or C3 . Under these conditions, nuclear division occurred normally but the furrow formation, which was induced by the contractile ring consisting of actomyosin just beneath the plasma membrane, was impaired . Comicroinjection of rho GDI with the GTP gamma S-bound form of rhoA p21 prevented the rho GDI action . Moreover, the sperm-induced cytoplasmic division of Xenopus embryos was inhibited by microinjection into the embryos of the rhoA p21 pre-ADP-ribosylated by C3 which might serve as a dominant negative inhibitor of endogenous rho p21 . These results indicate that rho p21 together with its regulatory proteins regulates the cytoplasmic division through the actomyosin system.

Jpn J Psychiatry Neurol, 1993 Mar, 47(1), 115 - 9
Complete remission of neuroleptic-induced Meige's syndrome by botulinum toxin treatment: a case report; Kurata K et al.; A botulinum A toxin injection has beneficial effects on patients suffering from facial and cervical spastic disorders . However, its effect almost completely disappears within three months . We have reported a case of a 23-year-old schizophrenic patient with severe neuroleptic-induced Meige's syndrome in whom botulinum toxin treatment exerted a marked effect which lasted more than 15 months after the final injection of botulinum toxin in spite of continuous neuroleptic medication . It is concluded that botulinum can be recommended as a treatment of choice in neuroleptic-induced Meige's syndrome.

J Cell Biol, 1993 Mar, 120(6), 1529 - 37
Inhibition of PMA-induced, LFA-1-dependent lymphocyte aggregation by ADP ribosylation of the small molecular weight GTP binding protein, rho; Tominaga T et al.; Botulinum C3 exoenzyme specifically ADP-ribosylates a group of ras-related small molecular weight GTP-binding proteins, rho, and inhibits their biological activity . Using this enzyme, we examined the function of rho in PMA-induced activation of lymphocyte function-associated antigen-1 (LFA-1) in a B lymphoblastoid cell line, JY . Northern blot analysis revealed that among the three rho genes, rhoA mRNA was predominantly expressed in JY cells . Consistently, only one {32P}ADP-ribosylated band was found when the lysate of the cells was subjected to ADP ribosylation by C3 exoenzyme . When the cells were cultured with C3 exoenzyme, this substrate was ADP-ribosylated in situ in a time- and concentration-dependent manner . Concomitant with this ADP ribosylation, PMA-induced LFA-1/intercellular adhesion molecule (ICAM)-1-dependent aggregation of JY cells was inhibited . This inhibition was blocked by prior treatment of the enzyme with an anti-C3 monoclonal antibody, and overcome by stimulation with higher concentrations of PMA . The C3 exoenzyme-induced inhibition was not affected by shaking of the cell suspension, while inhibition of aggregation by cytochalasin B was abolished by this procedure, suggesting that the inhibitory effect of the C3 exoenzyme treatment was not due to decrease in cell motility . The C3 exoenzyme treatment affected neither protein phosphorylation in JY cells before and after PMA stimulation, nor affected surface expression of LFA-1 and ICAM-1 . These results suggest that rhoA protein works downstream of protein kinase C activation linking PMA stimulation to LFA-1 activation and aggregation in JY cells.

Nippon Ganka Gakkai Zasshi, 1993 Feb, 97(2), 150 - 5
{Passive length-tension curves of extraocular muscles after botulinum toxin injection}; Okano M; Botulinum A toxin was injected into the superior rectus muscles of albino rabbits . Length-tension curves of detached superior rectus muscles were continuously measured with a length and tension measuring device developed by the author . The device consists of a strain gauge for measuring passive length-tension, and a position encoder equipped with an eddy current motor . The passive load of the muscle injected with Botulinum A toxin was significantly less than that of control muscle a week after the injection, however, it was greater than that of control muscle in 2 or 3 weeks after the injection . The compliance of the treated muscle was significantly less than that of control muscle at 3 weeks after the injection.

Ugeskr Laeger, 1993 Feb 1, 155(5), 284 - 9
{Torticollis treated with botulinum toxin}; Wulff CH et al.; The effect of botulinum toxin A treatment was studied in patients with idiopathic torticollis . Twenty patients with idiopathic torticollis received electromyographically guided intramuscular botulinum toxin A into the hyperactive neck muscles . In all, 48 treatments were given . The injections were repeated with intervals of 11 to 35 weeks (mean 18.7 weeks) . The grade of improvement was estimated subjectively using a visual analogue scale . An overall improvement of 55% compared with the status before treatment was found . The best result obtained in each individual patient varied from 20% to 84% (mean 66%) . The effect of the treatment began to wear off after eight to 22 weeks (mean 11.5 weeks) . The side effects consisted of short term dysphagia in two patients . The administration of botulinum toxin in idiopathic torticollis is a safe and beneficial treatment with few side effects . The EMG guidance of injections proved to be helpful as it restricted the injections into muscles with electromyographic hyperactivity, thereby economizing the amount of toxin given . It is doubtful if double-blind studies of botulinum toxin can be undertaken due to the marked muscular wasting and weakness caused by the injections.

Otolaryngol Head Neck Surg, 1993 Feb, 108(2), 135 - 40
Crystalline preparation of botulinum toxin type A (Botox): degradation in potency with storage; Gartlan MG et al.; Laryngeal injection of botulinum toxin type A is currently the most effective method of treating spasmodic dysphonia . Botox, a crystalline preparation of botulinum toxin type A, is the only toxin approved for clinical use in the United States and is packaged in vials of 100 mouse units (MU) . One MU corresponds to the calculated median lethal intraperitoneal dose (LD50) injected in mice . The logistic problems arising from the need for repeated injections of small amounts of Botox have been addressed by several investigators by refreezing unused Botox for use at a later time . Although FDA labeling recommends that Botox not used within 4 hours of reconstitution be discarded, data regarding degradation in potency after reconstitution and refreezing are not currently available . Using the LD50 Swiss-Webster mouse bioassay and statistical analysis by the Probit procedure, a 69.8% loss in potency was found when Botox was reconstituted, immediately frozen, and then assayed 2 weeks later (p < 0.0001) . Statistically significant degradation in potency was seen after refrigerator storage for 12 hours (p = 0.007), but not for 6 hours (p = 0.16) . Clinical implications regarding the dilution, use, and storage of Botox are discussed.

Muscle Nerve, 1993 Feb, 16(2), 181 - 7
Sustained focal effects of low-dose intramuscular succinylcholine; Walker FO et al.; We studied low-dose intramuscular succinylcholine in 9 subjects as part as an ongoing investigation of its potential to predict responses to botulinum toxin . We measured compound muscle action potentials (CMAPs) from the extensor digitorum brevis (EDB) muscles in each foot before and after intramuscular injections of 2.5 mg of succinylcholine into the EDB . Succinylcholine reduced mean CMAP amplitudes to 42% of baseline; the maximal reduction occurred at 19 +/- 6 (mean +/- standard deviation) minutes . Recovery to 73% of the baseline CMAP amplitude (approximately 50% recovery from block) occurred at 105 +/- 49 minutes after injection . Repetitive (train-of-four) stimulation at 2 Hz produced mild CMAP decrements (5-25%), but only during the recovery phase . Varying the succinylcholine concentrations (10, 20, or 50 mg/mL) while holding the total drug dose constant did not change the rate of onset or the extent of block . No systemic complications occurred . We conclude that: (1) 2.5 mg intramuscular succinylcholine can safely induce selective muscle weakness with a time course that differs from intravenously administered succinylcholine; and (2) further clinical studies comparing intramuscular succinylcholine and botulinum toxin are warranted.

Biochem Biophys Res Commun, 1993 Jan 29, 190(2), 470 - 4
Protease activity of botulinum neurotoxin type E and its light chain: cleavage of actin; DasGupta BR et al.; We demonstrate here for the first time a proteolytic activity of botulinum neurotoxin type E which is not expressed unless the single chain approximately 150 kDa neurotoxic protein is nicked into the dichain approximately 150 kDa neurotoxin . Actin was cleaved, in vitro, at multiple sites by the dichain neurotoxin and the N-terminal approximately 50 kDa light chain segment isolated from the dichain neurotoxin . The scissile peptide bonds of actin invariably contained Arg or Lys at the P1 site . Proteolytic activity of the isolated light chain and expression of this activity in the dichain form of the neurotoxin are consistent with the light chain's and the neurotoxin's intracellular actions--inhibition of neurotransmitter release.

Acta Otolaryngol Suppl, 1993, 504, 155 - 7
Botulinum toxin treatment for spasmodic dysphonia; Kobayashi T et al.; Effective treatment of adductor type spasmodic dysphonia with botulinum toxin injection is presented . Patients showed objective and/or subjective improvement in phonation . The beneficial effect lasted for approximately 3 months . An immediate complication is temporary hoarseness or aphonia, mainly due to diffusion of BT into the adjacent muscles . This is avoided by limiting the injection to one vocal fold only and by keeping the dose at less than 5 units . Insertion technique of the needles, such as percutaneous and laryngoscopically controlled techniques, are discussed.

Eur Neurol, 1993, 33(3), 199 - 203
Effectiveness of botulinum toxin in the treatment of spasmodic torticollis; Boghen D et al.; Thirty-two patients with spasmodic torticollis were assessed quantitatively for posture deformity, tremor and range of neck movement, and qualitatively for pain and global subjective disability . All patients were then treated with intramuscular botulinum toxin injections into appropriate neck muscles . Fifty-three treatments were administered using dosages of toxin in the range of 50-100 U per muscle . The maximum dose administered at a single sitting was 280 U . The progress of the patients was assessed during an 18-month period . Seventy-five percent of patients showed documented improvement in both subjective and objective parameters and were considered treatment successes . Pain improved in 65%, posture in 65%, tremor in 50% and range in 46% . The side effects that occurred were transient and included fatigue, dysphagia, neck weakness, hoarseness and local pain . This study demonstrates that treatment with botulinum toxin is of significant benefit for the majority of patients with spasmodic torticollis.

Toxicon, 1993 Jan, 31(1), 13 - 26
Identification of the site at which phospholipase A2 neurotoxins localize to produce their neuromuscular blocking effects; Simpson LL et al.; Experiments were conducted on mouse hemidiaphragm preparations using five phospholipase A2 neurotoxins of differing chain structures and antigenicities {notexin (one chain); crotoxin (two chains not covalently bound), beta-bungarotoxin (two chains covalently bound); taipoxin (three chains), and textilotoxin (five chains; one copy each of three chains and two copies of a fourth chain)} . Three clostridial neurotoxins (botulinum neurotoxin types A and B, and tetanus toxin) were used in comparison experiments . Phospholipase A2 neurotoxins produced concentration-dependent blockade of neuromuscular transmission . There was no obvious relationship between chain structure and potency, but there was an indication of a relationship between chain structure and binding . The binding of notexin was substantially reversible, the binding of crotoxin was slightly reversible, and the binding of beta-bungarotoxin, taipoxin and textilotoxin was poorly reversible . Experiments with neutralizing antibodies indicated that phospholipase A2 neurotoxins became associated with binding sites on or near the cell surface . This binding did not produce neuromuscular blockade . When exposed to physiological temperatures and nerve stimulation, bound toxin disappeared from accessibility to neutralizing antibody . This finding suggests that there was some form of molecular rearrangement . The two most likely possibilities are: (1) there was a change in the conformation of the toxin molecule, or (2) there was a change in the relationship between the toxin and the membrane . The molecular rearrangement step did not produce neuromuscular blockade . At a later time there was onset of paralysis; the amount of time necessary for onset of blockade was a function of toxin concentration . Phospholipase A2 neurotoxins were not antagonized by drugs that inhibit receptor-mediated endocytosis . In addition, phospholipase A2 neurotoxins did not display the pH-induced conformational changes that are typical of other endocytosed proteins, such as clostridial neurotoxins . However, phospholipase A2 neurotoxins were antagonized by strontium, and this antagonism was expressed against toxins that were free in solution and toxins that were bound to the cell surface . Limited antagonism was expressed after toxins had undergone molecular rearrangement, and no antagonism was expressed after toxin-induced neuromuscular blockade . The cumulative data suggest that phospholipase A2 neurotoxins are not internalized to produce their poisoning effects . These toxins appear to act on the plasma membrane, and this is the site at which they initiate the events that culminate in neuromuscular blockade.

Neurology, 1993 Jan, 43(1), 183 - 5
Botulinum toxin in the treatment of writer's cramp: a double-blind study; Tsui JK et al.; We treated 20 patients with writer's cramp in a double-blind, placebo-controlled study . Each patient received two treatments in tandem, one with botulinum-A toxin (BTX-A) injections and another with normal saline, separated by 3 months . Treatment order was randomized and unknown to the patient and physician . Patients were assessed before each treatment and 2 and 6 weeks after each treatment by objective measurements of pen control . Twelve patients had improvement in pen control after treatment with BTX-A, but only four had significant improvement in writing . BTX-A injections are effective in relieving symptoms in selected cases of writer's cramp, particularly in those with significant wrist-joint deviation.

Mov Disord, 1993, 8(1), 38 - 42
Quantitative measurement of cervical range of motion in patients with torticollis treated with botulinum A toxin; Dykstra D et al.; Improvement in cervical range of motion in patients with spasmodic torticollis by botulinum A toxin injection is difficult to objectively measure . Recently, a three-dimensional cervical range of motion system (EMROM) that measures primary as well as secondary cervical angles has been developed . This system uses an electromagnetic tracking system for data collection and a personal computer for analysis and graphic display of the data . We have tested the EMROM system and, from our results, believe that it can be used clinically to objectively and accurately measure cervical range of motion in patients who have spasmodic torticollis and who receive botulinum toxin injections.

Mov Disord, 1993, 8(1), 33 - 7
Time course of distant effects of local injections of botulinum toxin; Garner CG et al.; Botulinum toxin A (btx) is used to treat focal dystonias . From accidental intoxications it is known that btx can cause generalized pathologic single-fiber electromyography (SFEMG) findings . We monitored the onset and course of these disturbances in eight patients who received a small dose of btx (2-22 ng) for therapy of focal dystonias in the head/neck region for the first time via repeated SFEMG investigations at days 0, 3, 6, 9, 12, 28, and 56 . Recordings were performed in the extensor digitorum brevis muscle, and in two patients additionally in the tibialis anterior muscle . In six of these patients we found an increase of jitter and blocking . The onset of these changes was in the range of 3-13 days after injection . Fiber density showed a tendency to increase . There was no correlation between SFEMG findings and the dose of injected btx . Possible mechanisms for these observations may be either a very efficient local uptake and retrograde axonal transport via the spinal motor neurons or a systemic distribution via the blood circulation.

Eur Neurol, 1993, 33(4), 316 - 9
Botulinum toxin: preferred treatment for hemifacial spasm; Flanders M et al.; This paper retrospectively analyses the clinical data of 65 consecutive cases of hemifacial spasm . A benign etiology was clinically evident in the majority of cases . Neuroradiologic investigation ruled out serious intracranial pathology in atypical cases . Trials of oral medication in 26 patients were consistently unsuccessful . Fifty-one patients had repeated lid and facial muscle injections of botulinum toxin during a period of 8 years . The mean duration of effective relief from spasm following an initial treatment was 18.9 weeks . This effect did not diminish with repeated injections.

J Physiol Paris, 1993, 87(2), 107 - 15
Analysis of the structure and expression of the VAMP family of synaptic vesicle proteins; Trimble WS; VAMP/synaptobrevin proteins were first discovered as small integral membrane proteins in synaptic vesicles of vertebrates and invertebrates . At least two isoforms are expressed in the central nervous system of mammals in non-overlapping patterns . Biochemical studies have revealed that the VAMP synaptic vesicle proteins are the specific target in the presynaptic nerve terminal of botulinum B neurotoxin and tetanus toxin metalloendoprotease activities . The fact that these toxins rapidly and completely abrogate neurotransmission suggests that VAMP proteins play an essential role in this process . More recently, immunologically related proteins have been identified in non-neuronal cells such as adipocytes . In addition, molecular genetic studies of yeast secretion have identified VAMP-related proteins as playing important roles in vesicular transport between the endoreticulum and Golgi . Taken together, these results suggest that the VAMP proteins found on synaptic vesicles might represent specialized forms of proteins which participate in general aspects of cell membrane trafficking.

Otolaryngol Pol, 1993, 47(5), 464 - 7
{Treatment of facial spasm with botulinum toxin}; Pietruski J; Facialis spasmus (FS) is a chronic hyperkinesia primarily affecting patients in the 5th and 6th decades . Only muscles innervated by the facial nerve (FN) are involved . In the beginning there are only slight jerks, mostly in the region of the eyelid . In due course all facial muscles get involved . Spasms are worse when the patient is anxious, watching TV, working, driving and then restrict social activities . Because the etiology has not been known, a lot of methods of surgical and medical treatment were unsuccessful . Recently the cause has been clarified: FS is though to be caused by an irritation of this nerve by a blood vessel . The condition can be cured by an operation to separate the nerve and blood vessel . If an operation is thought to be inappropriate, treatment of FS may be with botulinum toxin (BT) . BT is used in very small doses (nanograms), and is injected in the area of the muscle that is affected . This weakens the muscle and helps to prevent a spasm.

Eye, 1993, 7 ( Pt 4), 550 - 3
Management of symptomatic latent nystagmus; Liu C et al.; Most patients with latent nystagmus are asymptomatic and do not require treatment . We discuss the management by botulinum toxin injection and surgery of five cases of latent nystagmus in which the patients suffered loss of visual acuity on certain manoeuvres as a consequence of an exacerbation of the nystagmus amplitude . The importance of eye movement recordings for accurate diagnosis is stressed and the investigative role of botulinum toxin injection is discussed . Extraocular muscle surgery is helpful in some cases of symptomatic latent nystagmus.

Exp Brain Res, 1993, 96(1), 77 - 82
Types and time course of the alterations induced in monkey blink movements by botulinum toxin; Porter JD et al.; The alterations induced in eyelid movement metrics subsequent to unilateral injections of botulinum toxin type A into the orbicularis oculi muscle were studied in chronic alert monkeys using the search coil technique . Botulinum toxin caused rapid paralysis of blinks in the treated eyelid . The amplitude and peak velocity of blinks generated by this eyelid remained at or below 20% of that of the fellow, untreated eyelid for 10-20 days . Blink amplitude gain increased linearly thereafter, attaining control values by 40-60 days after injection . Recovery of blink peak velocity was slower . The adaptive alterations in blink duration that were observed during the acute phase of toxin paralysis suggest that the mechanisms responsible for blink reflex plasticity may produce bilateral adjustments in eyelid function . Taken together, these data establish a quantitative data base that can be exploited in order to: (1) better understand the neural adaptive mechanisms that operate during eyelid movements and (2) allow quantitative comparisons between current treatment protocols that employ botulinum toxin and protocols that may lead to improvements in the treatment of chronic eyelid spasm (blepharospasm).

Exp Brain Res, 1993, 96(1), 39 - 53
Botulinum toxin paralysis of the orbicularis oculi muscle . Types and time course of alterations in muscle structure, physiology and lid kinematics; Horn AK et al.; In chronically prepared guinea pigs, we investigated the time course of botulinum toxin A's (Bot A) effect on the blink reflex by monitoring lid movements and EMG activity prior to and after Bot A injection into the orbicularis oculi muscle (OOemg), or after nerve crush of the zygomatic nerve . We correlated these alterations with the morphological changes of the orbicularis oculi (lid-closing) muscles of the same animals . After Bot A treatment there was a profound reduction of OOemg activity and blink amplitudes as well as a slowing of maximum blink down-phase velocity . Blink up-phases, however, remained unchanged . Gradual recovery of OOemg magnitude and blink amplitude started around day 6; a functioning blink reflex appeared on day 21, and full recovery of blink amplitude occurred by day 42 . Crushing the zygomatic branch of the facial nerve produced similar changes in blink parameters, but recovery was much more rapid (15 days) than for Bot A-treated guinea pigs . The morphological analysis demonstrated that Bot A produced a denervation-like atrophy in the orbicularis oculi . No fiber type-specific alterations were noted, and all muscle fiber types ultimately recovered, with no longstanding consequences of the transient denervation . Our findings support the notion that functional recovery was the result of preterminal and terminal axonal sprouting that subsequently re-established functional innervation . Moreover, differences between the present findings and those seen after injection of Bot A into the extraocular muscles strongly support the hypothesis that the composition in terms of muscle fiber type and the properties of the motor control system of a given muscle greatly influence both how the particular muscle responds to toxin injection, and how effective the toxin is in resolution of neuromuscular disorders that affect a particular muscle . The present findings were consistent with clinical observations that Bot A produces only temporary relief in patients with essential blepharospasm . It is likely that the efficacy of Bot A in treatment of blepharospasm could be improved by using agents that suppress terminal sprouting . The close correspondence of the changes in blink physiology between human patients and guinea pigs after Bot A treatment demonstrate that the guinea pig is an excellent model system for testing strategies to prolong the beneficial effects of Bot A treatment in relieving lid spasms in human subjects.

Ann Otolaryngol Chir Cervicofac, 1993, 110(3), 125 - 8
{Abnormal movements of the larynx . Diagnostic approach and therapeutic perspectives}; Angelard B et al.; Laryngeal movement disorders of neurological cause are actually misunderstood . In the course of chorea, or tardive post neuroleptic dyskinesia, vocal, swallowing for threatening breathing troubles could occur . Ten patients with generalized dyskinesias were studied by endoscopy with flexible laryngoscope and laryngeal electromyogram . Four of them were choreic and six had tardive dyskinesia (one had a severe breathing disorder) . Nine patients had abnormal movement disorders (MD) involving intrinsic laryngeal musculature . MD were spontaneous or were triggered by vocalization . These facts suggest that dyskinesia as they involved the upper airway tract could be life threatening . Laryngeal electromyography is a useful method for diagnosis and treatment of these dyskinesia . As in spasmodic dysphonia, botulinum toxin could be helpful in this local treatment.

Rev Neurol (Paris), 1993, 149(3), 202 - 6
{Hemifacial spasm treated with botulinum toxin}; Jedynak CP et al.; Botulinum toxin produces a transient presynaptic focal block at the neuromuscular junction . Thus it induces muscle weakness which has a significant beneficial effect on hemifacial spasm . Fifty-four patients were treated and received at least 2 and at the most 22 consecutive injections over a 5-year period . Injections were repeated every 9 weeks on average . The interval between injections corresponded to the time elapsed between the last injection and the re-emergence of spasms . Nineteen patients abandoned the treatment for various reasons . The most frequent side-effect was a ptosis which was observed in 1 out of 6 injections . Ptosis is due to diffusion of the botulinum toxin over a territory larger than expected . The results obtained in 42 patients were analyzed: there was no effect in 7 patients (17%); 11 patients improved by less than 50%; 13 by 50 to 70% and 11 more than 75%.

Ophthal Plast Reconstr Surg, 1993, 9(3), 182 - 90
Botulinum B toxin as an alternative to botulinum A toxin: a histologic study; Borodic GE et al.; Histochemical effects of botulinum B toxin were studied on fibers from longissimus dorsi muscle in Albino rabbits and compared to effects produced by botulinum A toxin . Acetylcholinesterase staining, muscle fiber size analysis, and ATPase staining indicated botulinum B toxin produced a denervation gradient and field similar to that produced by botulinum A toxin . At 5 weeks postinjection with botulinum B toxin, analysis showed muscle fiber size variability, and diffuse acetylcholinesterase fiber staining comparable to botulinum A toxin at the injection site . Muscle sections taken at 4.0 cm for analysis showed statistically significant decreased fiber size variability and contraction of acetylcholinesterase staining pattern for both immunotypes . In addition, the denervation reflected by histochemical staining and fiber size analysis appeared transient and lasted for approximately 3 months for both immunotypes . These findings suggest botulinum B toxin produces pharmacologic effects on innervation of striated muscle similar to botulinum A toxin . Because immunologic tolerance has been demonstrated after therapeutic botulinum A toxin injections, further clinical studies need to be conducted with other immunotypes of toxin with no cross-reactivity to type A.

Eur Arch Otorhinolaryngol, 1993, 250(5), 271 - 6
A comparison of bilateral and unilateral botulinum toxin treatments for spasmodic dysphonia; Zwirner P et al.; To assess the efficacy of bilateral or unilateral botulinum treatments for spasmodic dysphonia we injected botulinum toxin (type A) into the thyroarytenoid muscle of 24 patients with adductor type spasmodic dysphonia . Eleven patients underwent unilateral procedures and 13 bilateral procedures . Samples of sustained phonation were analyzed acoustically by a computer-assisted method and the air flow rates determined . All tests were conducted 1 week before injection and 1 week and 1 month after treatment . With unilateral injection, improvements in acoustic parameters occurred as early as 1 week after treatment . With bilateral injections, only the voice break factor was significantly reduced after 1 week, while standard deviations of fundamental frequency, jitter, shimmer and signal-to-noise-ratios were reduced 1 month after treatment . In comparison with unilateral injections, the mean air flow rate was twice as high 1 week after bilateral injections, with no significant differences found 1 month after treatment . Clinically, both injection modes resulted in the reduction of laryngeal spasms as early as within 48 h after injection.

Acta Otorhinolaryngol Belg, 1993, 47(3), 359 - 63
Botulinum A toxin injection in patients with facial nerve palsy; Smet-Dieleman H et al.; The preliminary experience of the authors with Botulinum toxin injections as a therapeutic tool for the ocular problems following facial nerve palsy is reported . Patients attained with facial nerve palsy who might benefit from Botulinum toxin were divided into three groups according to their eye problem: incomplete eyelid closure, postparalytic secondary facial spasm with synkinesis and contracture of the internal rectus muscle in the case of association with an abducens nerve palsy.

Bol Asoc Med P R, 1993 Jan-Mar, 85(1-3), 7 - 11
The use of botulinum toxin type A for the treatment of facial spasm; Serrano LA; We describe our experience with the use of botulinum A toxin for the treatment of patients with facial spasms . Thirty four patients with blepharospasm, thirty eight with hemifacial spasms and three with spastic entropion were injected with the use of Botulinum A toxin . Length of follow up ranged from 6 to 60 months . The effect of toxin lasted an average of 12.1 weeks in patients with blepharospasm and 15.5 weeks in patients with hemifacial spasms . This difference in mean response was statistically significant (p = 0.0001) . The most common side effect was ptosis and dry eyes . All side effects were transient in nature, lasting between three and twelve weeks . Botulinum toxin type A injections represent a good alternative for the treatment of facial spasms . It is a safe and effective office procedure . Most patients tolerate the procedure well . Its principal drawback is its transient effect and the need for repeated injections.

Bull Soc Belge Ophtalmol, 1993, 248, 77 - 82
Botulinum A toxin therapy in ophthalmology: a report of patients with facial dystonias; Vissenberg I et al.; An overview is given of the indications, results and complications with the use of botulinum A toxin injection therapy specifically in the treatment of facial dystonias . A total of 79 patients have been treated over a 5-year period . The patients were divided in four groups: essential blepharospasm, hemifacial spasm, apraxia of the opening of the eyes and spastic entropion . On average, the therapeutical effect started the third day after injection . The average duration of the beneficial effect was dependent of the underlying pathology . Local side effects were mild and systemic side effects were not seen . Botulinum A toxin therapy proved to have a reliable and reproducible effect in appropriate dosages, in 90% of the patients.

Rev Laryngol Otol Rhinol (Bord), 1993, 114(4), 281 - 7
{Indications for botulinum toxin in laryngectomy}; Klap P et al.; The botulinum toxin is a highly potent neurotoxin, used since several years in the treatment of the focal musculary dystonias . We define the laryngeal dystonia as a clinical entity, which is represented by a spasmodic dysphonia or an inspiratory dyspnea without dysphonia, related to a specific dystonia of the thyroarytenoid muscles . The laryngeal functional exploration (i.e . fibroscopy, videostroboscopy, acoustic analysis, computerized voice analysis), the neurological and electrophysiological assessment allow to make diagnosis and evaluation of the therapeutical results of this rare laryngeal neurologic disease which was relatively misunderstood until now . There are several clinical types of spasmodic dysphonia: adductor form (with a jerky voice, pitch beaks, vocal arrests and pneumophonatory incoordination); abductor form (with a breathy voice of very low intensity) and mixed types which can be difficult to identify . Since 1989, we have treated 55 laryngeal dystonias with local intra-muscular botulinum toxin injection: 48 spasmodic dysphonia and 7 inspiratory dyspnea without dysphonia, with hyperactivity of the thyroarytenoid muscles . We present our therapeutical protocol and results which are good in 87% of cases.

J Calif Dent Assoc, 1993 Jan, 21(1), 19 - 30
Oral motor disorders in humans; Clark GT et al.; Motor disorders affecting the orofacial musculature include bruxism, chronic orofacial muscle pain affecting the jaw and neck muscles and the involuntary waking period disorders such as orofacial dyskinesia, oral mandibular dystonia, tremor and others . Research at UCLA has touched these and many other areas . Current results have indicated the usefulness of contingent afferent electrical stimulation of the lip to control bruxism; provided information regarding the fatigue, endurance and recovery faculties of the protrusive jaw muscles; explored the issue of chronic muscle hyperactivity inducing headache pain; and worked with botulin toxin as a method to treat orofacial dystonia and dyskinesia.

Toxicon, 1992 Dec, 30(12), 1555 - 62
Inhibition of norepinephrine secretion from digitonin permeabilized PC12 cells by botulinum type D toxin; Yokosawa N et al.; Botulinum type D neurotoxin inhibited Ca(2+)-evoked norepinephrine secretion in digitonin permeabilized PC12 cells . Inhibition by the toxin required prior incubation with dithiothreitol (DTT) . The inhibition was dependent on both concentration and incubation times of the toxin, and was affected by Ca2+ concentration . With less than 0.7 microM Ca2+ almost complete inhibition was observed; however, above 0.7 microM, Ca2+ stimulated additional norepinephrine release in a dose-dependent manner.

Cutis, 1992 Dec, 50(6), 415 - 6
Psoriasiform eruption from intramuscular botulinum A toxin; Bowden JB et al.; Botulinum A toxin is used intramuscularly in the treatment of spastic neuromuscular disorders, strabismus, and laryngeal dystonia . The toxin has recently been reported as being useful for the cosmetic removal of glabellar furrows . The clinical effect of the toxin lasts four months or longer . Systemic side effects are rare and usually transient . We report the case of a psoriasiform eruption temporally related to the injection of botulinum A toxin into the medial rectus muscle to treat an ocular motility disorder . To our knowledge, this is the first case of a psoriasiform dermatitis caused by this agent.

Brain Res Bull, 1992 Dec, 29(6), 917 - 24
Adherence of botulinum and tetanus neurotoxins to synaptosomal proteins; Schengrund CL et al.; The ability of 125I-labeled botulinum type A and tetanus neurotoxins to adhere to blots of synaptosomal proteins separated by SDS-polyacrylamide gel electrophoresis was studied . Both neurotoxins appeared to adhere preferentially to an approximately 80 kDa and to a lesser extent to an approximately 116 kDa protein(s) . Adherence of the neurotoxins to these proteins was enhanced by preincubation of the neurotoxins with GT 1b . The approximately 100 kDa heavy chain segment of BTxA adhered to the same proteins . The carboxy terminal half of the heavy chain adhered primarily to the approximately 80 kDa protein(s) while the amino terminal portion bound most intensely to the approximately 116 kDa protein(s) . The ability of the approximately 80 and approximately 116 kDa proteins to stain positively with the periodic acid-Schiff reagent and to bind 125I-labeled wheat germ lectin suggests that they are glycosylated . Both neurotoxins appear to adhere to the same approximately 80 and approximately 116 kDa proteins because tetanus neurotoxin preincubated with GT 1b was able to reduce binding of radiolabeled botulinum type A neurotoxin to both proteins . Neither neurotoxin adhered to blots of proteins from liver, spleen, or kidney, suggesting that the proteins adhered to are neural components.

Neurology, 1992 Dec, 42(12), 2263 - 6
Hemimasticatory spasm: clinical and electrophysiologic observations; Auger RG et al.; Hemimasticatory spasm is a rare disorder of the trigeminal nerve that produces involuntary jaw closure due to paroxysmal unilateral contraction of jaw-closing muscles . We report three patients with this disorder . Electrophysiologic studies demonstrated normal blink and masseter reflexes . The masseter inhibitory reflex was absent during periods of spasm . Needle electromyography demonstrated irregular bursts of motor unit potentials that were identical to the pattern observed in hemifacial spasm . The electrophysiologic findings suggest ectopic excitation of the trigeminal motor root or its nucleus, an abnormality that is analogous to ectopic excitation of the facial nerve in hemifacial spasm . One patient improved temporarily with surgery, one improved while on treatment with carbamazepine, and another responded favorably to botulinum toxin injection.

Plast Reconstr Surg, 1992 Dec, 90(6), 972 - 7; discussion 978-9
Contralateral injections of botulinum A toxin for the treatment of hemifacial spasm to achieve increased facial symmetry; Borodic GE et al.; Six patients noted facial asymmetry after botulinum toxin injection for hemifacial spasm . Each patient was injected on the side contralateral to the spasms with 10 to 15 IU over the zygomatic major and minor muscles . Each patient noted improvement in facial symmetry in the resting position and dynamic facial movements . Five of the six patients desired this approach with subsequent injections . This injection method variation proved helpful in the managing of hemifacial weakness created by botulinum A toxin for this condition.

Yonsei Med J, 1992 Dec, 33(4), 289 - 93
Spasmodic torticollis: medical and botulinum A toxin treatment; Lee MC; The exact pathophysiologic mechanisms of spasmodic torticollis and other idiopathic torsion dystonias remain largely unknown . Thus, a variety of drugs have been used alone or in combination on an empirical basis to treat these disorders, but to date none have efficacy that is proven and consistent . The drugs in use include anticholinergics, benzodiazepines, dopaminergics and dopamine antagonists with variable degrees of clinical improvement . Botulinum toxin A injection treatment for spasmodic torticollis is safe and efficacious with minimal adverse effect . However, it is expensive and beneficial effects are short-lasting . Only when a spasmodic torticollis patient's symptoms are refractory to combined treatment, using various drugs and Botulinum toxin injections, should the patient be considered a candidate for neurosurgical procedures.

J Osaka Univ Dent Sch, 1992 Dec, 32, 27 - 44
Activation of cathepsin B involved in enkephalin production by bradykinin and its cleavage products in cultured fibroblasts of the rat dental pulp; Zhu BF et al.; Mechanisms of cathepsin B activation involved in methionine-enkephalin (ME) production induced by bradykinin (BK), des-Arg9-BK or L-arginine (L-Arg) were studied using cultured fibroblasts of the rat dental pulp, especially from a viewpoint of intracellular signal transduction . BK, des-Arg9-BK, L-Arg or cysteine enhanced the release of ME-like peptides from the cells, and the release of ME-like peptides induced by des-Arg9-BK was inhibited by des-Arg9-{Leu8}-BK (BK B1-receptor antagonist) and E-64 (a specific inhibitor of cysteine proteinases) . The activation of cathepsin B by BK or des-Arg9-BK was inhibited by des-Arg9-{Leu8}-BK or islet-activating protein (IAP), and the activation of cathepsin B by L-Arg was inhibited by Leu-Arg (kyotorphin-receptor antagonist) or Botulinum C3-enzyme . The activation of cathepsin B by those stimulants was dependent on calcium ion . These results suggest that the ME production by BK or des-Arg9-BK may be mediated by Ca(2+)-dependent cathepsin B activation through B1-receptors and IAP-sensitive G-proteins, whereas the production by L-Arg may be mediated by Ca(2+)-dependent cathepsin B activation through kyotorphin-receptor and Botulinum C3-enzyme-sensitive G-proteins . On the other hand, the activation of cathepsin B was inhibited by neomycin B (phospholipase C inhibitor) and various serine/threonine kinase inhibitors . These results indicate that phospholipase C and serine/threonine kinases are involved in the activation of cathepsin B by BK, des-Arg9-BK or L-Arg . Genistein inhibited the activation of cathepsin B by des-Arg9-BK or L-Arg in a different fashion, suggesting that tyrosine kinase(s) is also involved in the activation . Cathepsin B activation by BK or L-Arg but not des-Arg9-BK was inhibited by L-NMMA (inhibitor of NO synthesis), and the activation by L-Arg was enhanced by beta-glycerophosphate (beta-GP: inhibitor of phosphatases), while the activation by BK or des-Arg9-BK was inhibited by beta-GP . These results suggest that BK-induced cathepsin B activation in the fibroblasts may be due to a combined effect of des-Arg9-BK and L-Arg.

J Pharmacol Exp Ther, 1992 Dec, 263(3), 1269 - 74
Role of protein kinase C in short-term transmission at the mammalian neuromuscular junction; Considine RV et al.; Neuronal cells grown in culture were exposed to drugs that stimulate protein kinase C (phorbol myristate acetate), inhibit the catalytic site in protein kinase C (H7, staurosporine) or inhibit the regulatory site in protein kinase C (calphostin, sphingosine) . In NG-108 and N1E-115 cells, phorbol myristate acetate produced substantial stimulation of protein kinase C activity (0.1 microM produced approximately 75% stimulation) . In these same cells, H7 {100% inhibition concentration (IC100) approximately 1 mM} and staurosporine (IC100 approximately 0.2 microM) inhibited the catalytic site in the enzyme, and calphostin (IC80-IC90 approximately 2.0 microM) and sphingosine (IC80-IC90 approximately 1 microM) inhibited the regulatory site in the enzyme . Phorbol myristate acetate, as well as drugs that inhibit the catalytic and regulatory sites in protein kinase C, were tested for their effects on phrenic nerve-hemidiaphragm preparations . At concentrations that stimulated enzyme activity in neuronal cells in culture, phorbol myristate acetate did not augment normal transmission, nor did it restore transmission to preparations bathed in medium with low calcium (0.4-0.6 mM) . At concentrations equivalent to the IC80 to IC100 values in neuronal cells in culture, H7, staurosporine, calphostin and sphingosine did not paralyze short-term transmission, nor did they depress transmission in tissues bathed in low calcium . Pretreatment of neuromuscular preparations with phorbol myristate acetate, H7, staurosporine, calphostin or sphingosine did not alter the amount of time necessary for botulinum neurotoxin type A, botulinum neurotoxin type B or tetanus toxin to paralyze transmission . The data indicate that protein kinase C is not required for short-term neuromuscular transmission.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1992 Nov 25, 267(33), 23479 - 83
Botulinum neurotoxins are zinc proteins; Schiavo G et al.; The available amino acid sequences of 150-kDa botulinum and tetanus neurotoxins show the presence of a closely homologous segment in the middle of the light chain (NH2-terminal 50 kDa), which is the intracellularly active portion of the toxin . This segment contains the zinc binding motif of metalloendopeptidases, HEXXH . Atomic adsorption analysis of botulinum neurotoxins (serotypes A, B, and E) made on the basis of this observation demonstrated the presence of one zinc atom/molecule of 150-kDa neurotoxin . Conditions were found for the removal of the zinc ion with chelating agents and for the restoration of the normal metal content . The conserved segment, which includes the zinc binding motif, was synthesized and shown to bind {65Zn}2+ . Chemical modification experiments indicated that two histidines and no cysteines are involved in Zn2+ coordination in agreement with a probable catalytic role for the zinc ion . The present findings suggest the possibility that botulinum neurotoxins are zinc proteases.

FEBS Lett, 1992 Nov 16, 313(1), 12 - 8
Identification of an ion channel-forming motif in the primary structure of tetanus and botulinum neurotoxins; Montal MS et al.; Synthetic peptides with amino acid sequences corresponding to predicted transmembrane segments of tetanus toxin were used as probes to identify a channel-forming motif . A peptide denoted TeTx II, with sequence GVVLLLEYIPEITLPVIAALSIA, forms cation-selective channels when reconstituted in planar lipid bilayers . The single channel conductance in 0.5 M NaCl or KCl is 28 +/- 3 and 24 +/- 2 pS, respectively . In contrast, a peptide with sequence NFIGALETTGVVLLLEYIPEIT, denoted as TeTx I, or a peptide with the same amino acid composition as TeTx II but with a randomized sequence, do not form channels . Conformational energy calculations show that a bundle of four amphipathic alpha-helices is a plausible structural motif underlying observable pore properties . The identified functional module may account for the channel-forming activity of both tetanus toxin and the homologous botulinum toxin A.

Ann Neurol, 1992 Nov, 32(5), 633 - 42
Effectiveness of botulinum toxin administered to abolish acquired nystagmus; Leigh RJ et al.; We injected botulinum toxin into the horizontal rectus muscles of the right eyes of 2 patients who had acquired pendular nystagmus with horizontal, vertical, and torsional components . This treatment successfully abolished the horizontal component of the nystagmus in the injected eye in both patients for approximately 2 months . Both patients showed a small but measurable improvement of vision in the injected eye that may have been limited by coexistent disease of the visual pathways . The vertical and torsional components of the nystagmus persisted in both patients . In 1 patient, the horizontal component of nystagmus in the noninjected eye increased; we ascribe this finding to plastic-adaptive changes in response to paresis caused by the botulinum toxin . Such plastic-adaptive changes and direct side effects of the injections--such as diplopia and ptosis--may limit the effectiveness of botulinum toxin in the treatment of acquired nystagmus . Neither patient elected to repeat the botulinum treatment.

Ann Otol Rhinol Laryngol, 1992 Nov, 101(11), 888 - 92
Quantitative mapping of the effect of botulinum toxin injections in the thyroarytenoid muscle; George EF et al.; Spasmodic dysphonia has been successfully treated by thyroarytenoid muscle injections of botulinum toxin (Botox) with dosages ranging from 0.625 to 25 U . In some patients, excessive paralysis with resulting breathiness and aspiration have been noted . In order to maximize the efficiency of Botox injections, the histologic effects of various Botox dosages were examined in the dog . Nine canine thyroarytenoid muscles were injected with 0.5 to 12.5 U of Botox . After 24 hours, the recurrent laryngeal nerve to the injected muscle was electrically stimulated in order to deplete the glycogen within the muscle fibers . Frozen sections of this muscle were then stained for glycogen . Those fibers that retained their glycogen were presumed paralyzed by the Botox injection . The extent of paralysis was found to be dose-related from 1.0 to 7.5 U . At 10 U and above the muscle was completely paralyzed . Spread of the toxin to the lateral cricoarytenoid muscle was seen at doses as low as 1.0 U . Clearly, doses less than 10 U appear sufficient for clinical paralysis.

Ann Otol Rhinol Laryngol, 1992 Nov, 101(11), 883 - 7
Point-touch technique of botulinum toxin injection for the treatment of spasmodic dysphonia; Green DC et al.; Intralaryngeal injections of botulinum toxin (Botox), under electromyographic guidance, have emerged as an effective treatment for adductor spasmodic dysphonia . To remain effective, these injections must be repeated every 3 to 9 months as the symptoms recur . One drawback to the current method is the need for electromyographic confirmation of needle placement into the thyroarytenoid muscle . This report describes an anatomic approach to Botox injection that requires only flexible nasopharyngeal endoscopy and careful evaluation of the anatomic landmarks . This technique has been used successfully on 13 patients, and objective pretreatment and posttreatment measures are reported.

Otolaryngol Head Neck Surg, 1992 Nov, 107(5), 684 - 96
An evaluation of laryngeal muscle activation in patients with voice tremor; Koda J et al.; Eight patients with voice tremor were studied to characterize laryngeal muscle involvement . Electromyographic (EMG) recordings were made from intrinsic laryngeal muscles, simultaneously with some extrinsic laryngeal muscles, respiratory movement, and voice recordings during respiration, whisper, and phonation . Spectral measures were used to determine the tremor frequency and the prominence of spectral peaks in the EMG, respiratory and acoustic signals, while correlation coefficients were computed between pairs of tremulous EMG signals to measure the synchrony of tremor between muscles . The intrinsic laryngeal muscles were tremulous during respiration and speech, with the thyroarytenoid most often involved . Tremor was also detected in some of the extrinsic muscle recordings and the percentage of muscles with tremor was higher during phonation than during whisper or respiration . Time delays were found between tremor oscillations in laryngeal muscles . Because the thyroarytenoid was affected in all the patients studied, botulinum toxin injections may be beneficial in treatment of this voice disorder.

Otolaryngol Head Neck Surg, 1992 Nov, 107(5), 657 - 68
Bilateral thyroarytenoid denervation: a new treatment for laryngeal hyperadduction disorders studied in the canine; Sercarz JA et al.; Adductor spasmodic dysphonia is a vocal disorder of uncertain etiology with no satisfactory long-term treatment . Recently, injection of botulinum toxin (Botax) into the thyroarytenoid (TA) muscle has been used as an effective temporary treatment . A surgical counterpart to bilateral TA Botox injection is described in this article . Bilateral thyroarytenoid denervation was performed through a window in the thyroid cartilage in seven canines, including four that were studied 3 months after the procedure . No serious complications occurred in the animals, each maintaining full vocal fold abduction and adduction . In all cases, anticipated physiologic changes in laryngeal function were observed, including the inability to generate high subglottic pressures during high levels of laryngeal nerve (RLN) stimulation . In two of the surviving animals, the ansa cervicalis was used to reinnervate the TA muscle, thereby preventing the possibility of reinnervation from the proximal RLN stump while limiting TA atrophy and fibrosis . Bilateral TA denervation represents a hopeful new long-term approach to spasmodic dysphonia treatment.

J Neurosurg, 1992 Nov, 77(5), 808 - 9
Botulinum toxin enhancement of postoperative immobilization in patients with cervical dystonia . Technical note; Traynelis VC et al.; Postoperative immobilization in patients with cervical dystonia requiring fusion presents a unique management problem . Two patients with severe degenerative cervical spine disease secondary to chronic repetitive motion are reported . Both required a surgical fusion and postoperative immobilization . Botulinum toxin was injected intramuscularly to assist in immobilization . The technique used is described.

Infect Immun, 1992 Nov, 60(11), 4648 - 55
Visualizations of binding and internalization of two nonlinked protein components of botulinum C2 toxin in tissue culture cells; Ohishi I et al.; Binding and internalization of two nonlinked components of botulinum C2 toxin were visualized in tissue culture cells with components directly labeled with fluorescence . The binding of both untrypsinized and trypsinized component II (UT-II and T-II, respectively) to common specific sites on the cell membrane was evidenced by competitive binding between fluorescence-labeled and unlabeled components . The distribution patterns of fluorescence-labeled T-II and UT-II after binding to cells at 37 degrees C were different; T-II clustered on the cell membrane and entered the cells in endosomes, whereas UT-II entered the cells inefficiently and not in vesicles and was distributed on the nuclear surface . The difference may be due to the multivalent property of T-II, which is not shared with UT-II . Fluorescence-labeled component I, which binds only to cells bound with T-II, entered cells by the same route as T-II did; both colocated on the same clusters on the cell membrane and also in the same vesicles in the cytoplasm . The present results suggest that component I of C2 toxin, which ADP-ribosylates cytoplasmic actin, directly binds to T-II but not to UT-II on the cell membrane and is internalized into cells together with T-II in the same endosomes.

Eksp Klin Farmakol, 1992 Nov-Dec, 55(6), 46 - 8
{The nonspecific mechanisms of the pharmacological correction of motoneuron trophic failure in experimental botulin poisoning}; Mikhailov VV et al.; Cobalamines, leukovorine, cymidine phosphocholine, dalargin are pharmacological agents that affect the support of motoneuronal trophic functions, unlike the actoprotector bemityl, prevent the development of fatal botulinic intoxication, but have no effects on the specific mechanisms of botulinic damage when the threshold concentrations of toxin achieve target cells . The depotentiating effect of the drugs is associated with the increased natural degradation of botulinic toxin in its target cells.

Neurol Neurochir Pol, 1992 Nov-Dec, 26(6), 783 - 9
{Botulin in the treatment of local dystonia}; Domzal T; Botulin A has been introduced for the treatment of local dystonia especially blepharospasm and torticollis . Three cases of blepharospasm and 5 cases of torticollis were treated with botulin injections directly into the muscles by a method presented in detail . Good effects were obtained in blepharospasm but very poor in torticollis, which may have been due to too low doses of the toxin and inadequate choice of injection points . The method is safe and in only 1 case transient weakness of the masseters was noted.

Nature, 1992 Oct 29, 359(6398), 832 - 5
Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin; Schiavo G et al.; Clostridial neurotoxins, including tetanus toxin and the seven serotypes of botulinum toxin (A-G), are produced as single chains and cleaved to generate toxins with two chains joined by a single disulphide bond (Fig . 1) . The heavy chain (M(r) 100,000 (100K)) is responsible for specific binding to neuronal cells and cell penetration of the light chain (50K), which blocks neurotransmitter release . Several lines of evidence have recently suggested that clostridial neurotoxins could be zinc endopeptidases . Here we show that tetanus and botulinum toxins serotype B are zinc endopeptidases, the activation of which requires reduction of the interchain disulphide bond . The protease activity is localized on the light chain and is specific for synaptobrevin, an integral membrane protein of small synaptic vesicles . The rat synaptobrevin-2 isoform is cleaved by both neurotoxins at the same single site, the peptide bond Gln 76-Phe 77, but the isoform synaptobrevin-1, which has a valine at the corresponding position, is not cleaved . The blocking of neurotransmitter release of Aplysia neurons injected with tetanus toxin or botulinum toxins serotype B is substantially delayed by peptides containing the synaptobrevin-2 cleavage site . These results indicate that tetanus and botulinum B neurotoxins block neurotransmitter release by cleaving synaptobrevin-2, a protein that, on the basis of our results, seems to play a key part in neurotransmitter release.

J Biol Chem, 1992 Oct 25, 267(30), 21338 - 43
Tetanus toxin and botulinum toxins type A and B inhibit glutamate, gamma-aminobutyric acid, aspartate, and met-enkephalin release from synaptosomes . Clues to the locus of action; McMahon HT et al.; Tetanus toxin (100 nM) when preincubated with guinea pig cerebrocortical synaptosomes for 45 min reduces the final extent of the KCl-evoked, Ca(2+)-dependent, glutamate transmitter release to 30% of non-intoxicated controls . Similarly, 100 nM Botulinum neurotoxins, types A and B, preincubated for 90 min inhibit release to 45-60% of non-intoxicated controls . The toxins preferentially attenuate a slow phase of KCl-evoked glutamate release which may be associated with synaptic vesicle mobilization . Tetanus toxin additionally inhibits the release of aspartate, gamma-aminobutyric acid and met-enkephalin from the same preparation . Since amino acids and neuropeptides are released by distinct mechanisms, this indicates that the toxin affects a step common to both exocytotic pathways . When Ba2+ (which does not interact with calmodulin) is substituted for Ca2+, the control KCl-evoked release of each transmitter is unaffected and tetanus toxin is still inhibitory . Taken together these results implicate a calmodulin-independent locus (or loci) of action common to small- and large-dense-core vesicles and associated with vesicle transport.

J Biol Chem, 1992 Oct 15, 267(29), 20921 - 6
A rho gene product in human blood platelets . II . Effects of the ADP-ribosylation by botulinum C3 ADP-ribosyltransferase on platelet aggregation; Morii N et al.; In the accompanying paper (Nemoto, Y., Namba, T., Teru-uchi, T., Ushikubi, F., Morii, N., and Narumiya, S . (1992) J . Biol . Chem . 267, 20916-20920), we have identified rhoA protein as the sole substrate protein for botulinum C3 ADP-ribosyltransferase (C3 exoenzyme) in human blood platelets . Here we examined the role of rhoA protein in platelet functions . C3 exoenzyme added to washed platelets dose- and time-dependently ADP-ribosylated rhoA protein in situ in the cells . Concomitant with this modification, inhibition of thrombin-induced platelet aggregation was observed . This inhibition was not reversed by washing the treated platelets, but was not found when C3 exoenzyme was pretreated with mouse monoclonal anti-C3 exoenzyme antibody . C3 exoenzyme treatment did not affect thrombin-induced inositol 1,4,5-trisphosphate production . Secretion of preloaded {14C}serotonin was delayed by the enzyme treatment, but the extent of the secretion was not influenced . In addition, the enzyme treatment did not change the expression of the glycoprotein IIb-IIIa complex on the platelet surface . The enzyme treatment also suppressed platelet aggregation induced by phorbol myristate acetate . These results suggest that rhoA protein plays a role mainly in the aggregation process downstream from receptor-phospholipase C coupling . This, together with the previous finding that rhoA protein modulates stress fiber formation in cultured fibroblasts (Paterson, H . F., Self, A . J., Garrett, M . D., Just, I., Aktories, K., and Hall, A . (1990) J . Cell Biol . 111, 1001-1007), suggests that rhoA protein regulates the assembly of actin filaments and the avidity of the platelet integrin (glycoprotein IIb-IIIa) in the aggregation process.

J Biol Chem, 1992 Oct 15, 267(29), 20916 - 20
A rho gene product in human blood platelets . I . Identification of the platelet substrate for botulinum C3 ADP-ribosyltransferase as rhoA protein; Nemoto Y et al.; A substrate protein for botulinum C3 ADP-ribosyltransferase (C3 exoenzyme) in human platelets was purified to apparent homogeneity from the cytosol by ammonium sulfate fractionation and successive chromatography on columns of DEAE-Sepharose, hydroxylapatite, phenyl-Sepharose, and TSK phenyl-5PW . The purified protein yielded an amino acid sequence identical to that of rhoA protein . When platelet cytosol and membranes were incubated with C3 exoenzyme and {32P}NAD and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing, they gave only one {32P}ADP-ribosylated band on each electrophoresis that showed an M(r) of 22,000 and a pI of 6.0 . The radioactive bands from the two fractions co-migrated with each other and with the {32P}ADP-ribosylated purified protein . When these radioactive products were partially digested with either alpha-chymotrypsin or trypsin and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the same digestion pattern was found in the three samples . These results suggest that the ADP-ribosylation substrate for C3 exoenzyme in the platelet cytosol and membrane is rhoA protein and that it is the sole substrate detectable in human platelets.

Br J Urol, 1992 Oct, 70(4), 387 - 9
Botulinum toxin in the treatment of chronic urinary retention in women; Fowler CJ et al.; Six women were identified as having difficulty in voiding or complete urinary retention due to abnormal myotonic-like electromyographic (EMG) activity in the striated muscle of the urethral sphincter . An attempt was made to improve voiding by injection of botulinum toxin into the striated sphincter muscle . Although 3 patients then developed transient stress incontinence, demonstrating that sufficient botulinum toxin had been given to cause sphincter weakness, no patient had significant symptomatic benefit.

Appl Environ Microbiol, 1992 Oct, 58(10), 3426 - 8
Stabilization of botulinum toxin type A during lyophilization; Goodnough MC et al.; Botulinum toxin for medical use is diluted to very low concentrations (nanograms per milliliter); when it is preserved by lyophilization, considerable loss of activity can occur . In the present study, conditions that gave > 90% recovery of the toxicity after lyophilization of solutions containing 20 to 1,000 mouse 50% lethal doses per ml were found . Toxicity was recovered upon drying 0.1 ml of toxin solution when the pH was maintained below 7 and bovine or human serum albumins were used as stabilizers . Various other substances tested with albumin, including glucose, sucrose, trehalose, mannitol, glycine, and cellibiose, did not increase recovery on drying.

EMBO J, 1992 Oct, 11(10), 3577 - 83
Tetanus toxin is a zinc protein and its inhibition of neurotransmitter release and protease activity depend on zinc; Schiavo G et al.; Tetanus and botulinum neurotoxins are the most potent toxins known . They bind to nerve cells, penetrate the cytosol and block neurotransmitter release . Comparison of their predicted amino acid sequences reveals a highly conserved segment that contains the HexxH zinc binding motif of metalloendopeptidases . The metal content of tetanus toxin was then measured and it was found that one atom of zinc is bound to the light chain of tetanus toxin . Zinc could be reversibly removed by incubation with heavy metal chelators . Zn2+ is coordinated by two histidines with no involvement in cysteines, suggesting that it plays a catalytic rather than a structural role . Bound Zn2+ was found to be essential for the tetanus toxin inhibition of neurotransmitter release in Aplysia neurons injected with the light chain . The intracellular activity of the toxin was blocked by phosphoramidon, a very specific inhibitor of zinc endopeptidases . Purified preparations of light chain showed a highly specific proteolytic activity against synaptobrevin, an integral membrane protein of small synaptic vesicles . The present findings indicate that tetanus toxin, and possibly also the botulinum neurotoxins, are metalloproteases and that they block neurotransmitter release via this protease activity.

Schweiz Med Wochenschr, 1992 Sep 5, 122(36), 1311 - 6
{Treatment of movement disorders using botulinum toxin}; Burgunder JM; Intramuscular injections of botulinum toxin (Botox) are followed by a dose-dependent focal paresis which can be used to treat several focal movement disorders . Botox injections are recommended as effective for the treatment of blepharospasm, hemifacial spasm, and cervical dystonia (torticollis) . Focal dystonias elsewhere (for example, writer's cramp) can often be treated with similar success . Others, such as oromandibular dystonia, are more difficult to treat . In the case of more generalized dystonias, some focal muscle spasms can be treated with success by local intramuscular injections . New indications are still being investigated, for example in focal tremors and spasticity . Side effects are in general slight and disappear at the end of toxin effect . In general, it is necessary to repeat the injections after a couple of months, due to a cessation of effect after regrowth of nerve terminals . New injections have similar effects even over years of treatment.

Muscle Nerve, 1992 Sep, 15(9), 1045 - 9
Treatment of hemifacial spasm with botulinum toxin; Yoshimura DM et al.; The effectiveness of botulinum toxin injections in 11 patients with hemifacial spasm was investigated in a prospective placebo-controlled blinded study . The patients were treated with four sets of injections to various facial muscles, selected by clinical evaluation . Three injections were with graded doses of toxin and one was with placebo . The order of injections was random and unknown to the patients . Results were scored both subjectively by patient assessment of symptoms and objectively by blinded review of videotapes made one month after each injection . Subjective improvement occurred after 79% of injections with botulinum toxin, regardless of dose of toxin . Only 1 patient improved after placebo . Objective improvement was seen after 84% of injections with botulinum toxin . No patient showed objective improvement after placebo injection . The most frequent side effect was facial weakness, seen after 97% of injections of botulinum toxin . Facial bruising (20%), diplopia (13%), ptosis (7%), and various other mild side effects were seen less frequently . Botulinum toxin appears to be an effective and safe method of therapy for hemifacial spasm.

J R Soc Med, 1992 Sep, 85(9), 524 - 9
Botulinum toxin treatment of spasmodic torticollis; Anderson TJ et al.; We reviewed the efficacy and adverse effects of repeated botulinum toxin injections into hyperactive neck muscles of 107 successive patients with spasmodic torticollis . They received 510 injection treatments over a median period of 15 months (range 3-42 months) . One patient failed to benefit at all, but 101 (95%) patients reported considerable (moderate or excellent) benefit from at least one treatment . On a global subjective response rating, 93% of 429 treatments resulted in some improvement and 76% in moderate or excellent improvement . Pain reduction followed 89% of 190 treatments with moderate or excellent reduction after 66% . Median duration of benefit was 9 weeks . All torticollis types responded equally well and injections into two (or more) involved neck muscles were more effective than injection into a single muscle . The most frequent adverse effect was dysphagia, occurring after 44% of all treatments, but this was severe after only 2% . Antibodies to botulinum toxin were detected in the serum of three out of the five patients in whom loss of treatment efficacy occurred . We conclude that botulinum toxin treatment is the most effective available therapy for spasmodic torticollis and practical advice is provided for anyone wishing to set up the technique.

Naunyn Schmiedebergs Arch Pharmacol, 1992 Sep, 346(3), 358 - 61
Evidence for a link between specific proteolysis and inhibition of {3H}-noradrenaline release by the light chain of tetanus toxin; Sanders D et al.; The light chain of tetanus toxin is known to inhibit the Ca(2+)-evoked release of {3H}-noradrenaline from digitonin-permeabilized bovine adrenomedullary cells in culture but does not change the basal outflow or the total cellular radioactivity . Evidence for the involvement of proteolysis in this effect was obtained by three approaches . First, the permeabilized cells were exposed to a series of enzymes . The endoproteinase Glu-C mimicked the inhibition produced by the light chain . Second, protease inhibitors of different specificities were assessed for blockade of the action of light chain on {3H}-noradrenaline release from permeabilized cells . Blockade was complete with EDTA (2.5 mmol/l) or 1,10-o-phenanthroline (1 mmol/l), and absent with the highest concentrations tested of diisopropylfluorophosphate, phenylmethylsulfonyl fluoride, pepstatin, leupeptin, bestatin, phosphoramidon, thiorphan or trans-epoxysuccinic acid (E64) which is regarded as an inhibitor of thiol proteases . This inhibitor spectrum suggested that light chain might be a metalloprotease . Finally a sequence-His-Glu-Leu-X-His-occurring in the light chains of tetanus toxin and of the botulinum neurotoxins A, C, D, E was also found in many endoproteinases and an aminopeptidase . The motif is known to constitute their active site and to bind Zn2+ . In fact Zn2+ (0.6-0.9 mol/mol) was found in thoroughly dialysed two-chain tetanus toxin . The three approaches jointly support the hypothesis that the light chain of tetanus toxin, and probably of all clostridial neurotoxins, inhibits {3H}-noradrenaline release from adrenomedullary cells by degradation of (a) specific, still unknown protein(s) involved in exocytosis.

FEMS Microbiol Immunol, 1992 Sep, 5(1-3), 101 - 11
Ion channel and membrane translocation of diphtheria toxin; Montecucco C et al.; Diphtheria toxin is the best studied member of a family of bacterial protein toxins which act inside cells . To reach their cytoplasmic targets, these toxins, which include tetanus and botulinum neurotoxins and anthrax toxin, have to cross the hydrophobic membrane barrier . All of them have been shown to form ion channels across planar lipid bilayer and, in the case of diphtheria toxin, also in the plasma membrane of cells . A relation between the ion channel and the process of membrane translocation has been suggested and two different models have been put forward to account for these phenomena . The two models are discussed on the basis of the available experimental evidence and in terms of the focal points of difference, amenable to further experimental investigations.

J Neurol Neurosurg Psychiatry, 1992 Sep, 55(9), 844 - 5
Botulinum toxin therapy: distant effects on neuromuscular transmission and autonomic nervous system; Girlanda P et al.; To evaluate distant effects of botulinum toxin, single fibre electromyography on the extensor digitorum communis muscle and six tests of cardiovascular reflexes were performed in five patients injected with BoTox (Oculinum(R) 20-130 units) for craniocervical dystonia and hemifacial spasm . Patients underwent two sessions of treatment and the second time the dosage was doubled . Botulinum toxin injection induced an increase of mean jitter value above normal limits in all cases . An increase of fibre density was recorded six weeks after the treatment . Cardiovascular reflexes showed mild abnormalities in four patients . The data confirm distant effects of botulinum toxin on neuromuscular transmission and on autonomic function.

Arq Neuropsiquiatr, 1992 Sep, 50(3), 387 - 90
{Hemifacial spasm and basilar impression associated with Arnold-Chiari deformity . Report of a case}; Leal Filho MB et al.; The authors report a case of symptomatic basilar impression and Arnold-Chiari malformation being presented as the first symptom of hemifacial spasm . The surgical treatment of the malformation resulted in improvement of the clinical manifestation with reduction of the hemifacial spasm . The need for the aetiological therapy for the hemifacial spasm is emphasized, before symptomatic treatment with botulinum toxin is tried.

Tidsskr Nor Laegeforen, 1992 Aug 30, 112(20), 2660 - 2
{Botulinum toxin treatment of spastic torticollis}; Kerty E et al.; Botulinum toxin A was administered to 19 patients with spasmodic torticollis . A significant decrease of abnormal head and neck movements was recorded, and all the patients who suffered pain reported relief . Side effects were minor and transient . The results of this study indicate that botulinum toxin is an effective means of treating torticollis.

Eur J Cell Biol, 1992 Aug, 58(2), 259 - 70
Characteristics of pancreatic exocrine secretion produced by venom from the Brazilian scorpion, Tityus serrulatus; Fletcher PL Jr et al.; The influence of venom (TSV) from the Brazilian scorpion, Tityus serrulatus, on exocrine pancreatic secretion was studied in relation to known cholinergic and peptidergic secretagogue activity . Pulse-labeling followed by chase incubation in the presence of secretagogues and various pharmacological agents revealed unique physiological characteristics of TSV in guinea pig pancreatic lobules . Exocytotic discharge of newly synthesized 3H-labeled proteins during a 3-h chase incubation showed a marked increase over basal discharge levels using logarithmic TSV doses of 0.10 to 100 micrograms/ml . This stimulation was comparable to maximal values elicited by carbachol, cholecystokinin-octapeptide (CCK-8) or caerulein and discharge kinetics were similar . TSV-mediated secretion was ATP and calcium dependent and partially inhibited by atropine . Only tetrodotoxin completely blocked TSV stimulation of newly synthesized protein discharge . Both botulinum toxin and curare had no effect on venom stimulation, indicating that TSV interaction with exocrine pancreatic cells occurs postsynaptically . Verapamil, a calcium channel antagonist, produced a moderate inhibition of TSV stimulation . When antagonists to the cholecystokinin (CCK) receptor were incubated with TSV, no change in secretory activity occurred . Therefore, TSV does not bind to CCK receptors and probably operates through its own receptor which may be an ion channel . Additionally, morphological studies in vitro revealed a high level of pancreatic secretory activity as evidenced by dense secretory acinar luminal content, reduction in zymogen granule (ZG) population, and development of exocytotic images.

J Ark Med Soc, 1992 Aug, 89(3), 133 - 4
Treatment of cervical dystonia (torticollis) in adults with botulinum A toxin; Metzer WS et al.; Dystonias can be classified by etiology (idiopathic or symptomatic), by age of onset (childhood, adolescence or adult), and by anatomical involvement (focal, segmental or generalized) . Cervical dystonia (torticollis) is one of the most common focal dystonias . We describe our experience in the treatment of 15 consecutive cervical dystonia patients by chemodenervation with botulinum A toxin (BOTOX), with significant improvement being objectively measured . Botox is accepted as a safe and efficacious modality for the treatment of cervical dystonia.

Clin Neuropharmacol, 1992 Aug, 15(4), 276 - 88
Baclofen in the treatment of dystonia; Greene P; Dystonia refers to involuntary, prolonged muscle contractions leading to sustained, often twisting, postures . High dose anticholinergic therapy for childhood onset dystonia, botulinum toxin injections for focal dystonia, and levodopa for diurnal dystonia provide symptomatic relief for some patients . Despite this, treatment of both idiopathic and secondary dystonia remains inadequate for many patients . Baclofen, a pre-synaptic acting GABA agonist, has been reported to benefit dystonia in a number of retrospective studies . Dramatic improvement in symptoms, especially in gait, was found in almost 30% of 31 children and adolescents with idiopathic dystonia in one retrospective study using doses ranging from 40 to 180 mg daily . The response to baclofen of adults with focal dystonia is less dramatic . One series of 60 adults with cranial dystonia found sustained benefit in 18% . Smaller series have not consistently found significant benefit in adults . Baclofen has been used to treat several secondary dystonias: tardive dystonia has occasionally been reported to improve and there are isolated reports of improvement in dystonia occurring in Parkinson's disease and in glutaric aciduria.

J Neurol, 1992 Aug, 239(7), 375 - 8
Local botulinum toxin in the treatment of spastic drop foot; Dengler R et al.; Ten patients with spastic drop foot were treated by local injections of botulinum toxin A (botulinum toxin A haemagglutinin complex) . The purpose was to improve stance and gait and/or to facilitate physiotherapy and patient care . Various calf muscles were injected using EMG guidance . The average dose used was 23 ng . Prior to and 4 weeks after treatment, positions of the upper and lower ankle joint at rest and the corresponding end positions of passive and active movement were determined . In addition, changes of spasticity and pain, the transmission phenomenon and stance and gait were evaluated . Most patients showed improvement of some aspects of the spastic drop foot . Positions of the upper and lower ankle joint were improved in most of the patients, as were the other parameters examined . Except for weakness of the injected muscles no side-effects were observed . The results appear promising and may be optimized in further trials by using higher doses of the toxin.

J Biol Chem, 1992 Jul 25, 267(21), 14721 - 9
Minimal essential domains specifying toxicity of the light chains of tetanus toxin and botulinum neurotoxin type A; Kurazono H et al.; To define conserved domains within the light (L) chains of clostridial neurotoxins, we determined the sequence of botulinum neurotoxin type B (BoNT/B) and aligned it with those of tetanus toxin (TeTx) and BoNT/A, BoNT/C1, BoNT/D, and BoNT/E . The L chains of BoNT/B and TeTx share 51.6% identical amino acid residues whereas the degree of identity to other clostridial neurotoxins does not exceed 36.5% . Each of the L chains contains a conserved motif, HExxHxxH, characteristic for metalloproteases . We then generated specific 5'- and 3'-deletion mutants of the L chain genes of TeTx and BoNT/A and tested the biological properties of the gene products by microinjection of the corresponding mRNAs into identified presynaptic cholinergic neurons of the buccal ganglia of Aplysia californica . Toxicity was determined by measurement of neurotransmitter release, as detected by depression of postsynaptic responses to presynaptic stimuli (Mochida, S., Poulain, B., Eisel, U., Binz, T., Kurazono, H., Niemann, H., and Tauc, L . (1990) Proc . Natl . Acad . Sci . U . S . A . 87, 7844-7848) . Our studies allow the following conclusions . 1) Residues Cys439 of TeTx and Cys430 of BoNT/A, both of which participate in the interchain disulfide bond, play no role in the toxification reaction . 2) Derivatives of TeTx that lacked either 8 amino- or 65 carboxyl-terminal residues are still toxic, whereas those lacking 10 amino- or 68 carboxyl-terminal residues are nontoxic . 3) For BoNT/A, toxicity could be demonstrated only in the presence of added nontoxic heavy (H) chain . A deletion of 8 amino-terminal or 32 carboxyl-terminal residues from the L chain had no effect on toxicity, whereas a removal of 10 amino-terminal or 57 carboxyl-terminal amino acids abolished toxicity . 4) The synergistic effect mediated by the H chain is linked to the carboxyl-terminal portion of the H chain, as demonstrated by injection of HC-specific mRNA into neurons containing the L chain . This finding suggests that the HC domain of the H chain becomes exposed to the cytosol during or after the putative translocation step of the L chain.

Am J Ophthalmol, 1992 Jul 15, 114(1), 72 - 80
Superior rectus muscle overaction after cataract extraction; Grimmett MR et al.; Four patients with an ipsilateral hypertropia after cataract extraction consistent with superior rectus muscle overaction were identified between March 1990 and April 1992 . Operative trauma was the most likely causative factor, as other likely conditions were excluded . The proposed pathogenesis for all cases is similar to that of botulinum type-A toxin therapy: a transient postoperative weakness of the ipsilateral inferior rectur muscle leads to a contracture or strengthening of the ipsilateral antagonist (the superior rectus muscle) . Possible mechanisms of injury that would result in a transient inferior rectus muscle palsy would include anesthetic myotoxicity or direct trauma to the muscle and related structures from the retrobulbar injection (or subconjunctival injection) . Surgical intervention consisting of an ipsilateral superior rectus muscle recession and posterior fixation sutures (when the vertical incomitance was large) yielded excellent results in restoring single binocular vision . Possible preventive measures would include using a minimal volume of anesthetic along with careful needle placement.

Biochem J, 1992 Jul 1, 285 ( Pt 1), 9 - 12
Surface topography of histidine residues of tetanus toxin probed by immobilized-metal-ion affinity chromatography; Rossetto O et al.; Tetanus toxin contains 14 histidine residues: six of them are localized in the light chain (L), one is present in the N-terminal half of the heavy chain (HN) and the remaining seven histidines are localized in the C-terminal half of the heavy chain (Hc) . Using immobilized-metal-ion affinity chromatography with Chelating Superose-Zn(II), we show that histidines of Hc are exposed to the protein surface and are responsible for the binding of tetanus toxin and of Hc to the immobilized metal . The histidines of the L chain are not available for co-ordination of matrix-bound Zn2+; however, two of them and three of the histidines of fragment Hc are accessible to diethyl pyrocarbonate . Chromatography on Superose-Zn(II) is also shown to be a simple and efficient method for the rapid isolation of tetanus toxin and of its Hc fragment, which can be extended to the botulinum neurotoxins.

Neurology, 1992 Jul, 42(7), 1391 - 3
Botulinum toxin treatment of supranuclear ocular motility disorders; Newman NJ et al.; We treated one patient with bilateral internuclear ophthalmoplegia and another with skew deviation with extraocular muscle botulinum toxin injection . Both patients had pre-injection symptomatic diplopia in primary position, one for 1 month and the other for 12 months . Resolution of diplopia and complete and permanent binocular fusion in primary position was obtained within 3 to 4 days in both cases.

Neurology, 1992 Jul, 42(7), 1307 - 10
Dysphagia after botulinum toxin injections for spasmodic torticollis: clinical and radiologic findings; Comella CL et al.; We prospectively evaluated the frequency, severity, and radiologic features of swallowing abnormalities following Botox treatment of spasmodic torticollis . We performed both clinical and radiologic evaluations of swallowing before and following Botox in 18 consecutive cervical dystonia patients receiving their first Botox treatment . Before Botox, 11% of the patients had clinical symptoms of dysphagia and 22% had radiologic signs of a peristaltic abnormality . After Botox, the signs and symptoms of dysphagia in these patients did not change, but an additional 33% developed new dysphagic symptoms and 50% of the patients developed new peristaltic abnormalities by radiologic studies . Complaints of swallowing difficulty were always associated with abnormal radiologic findings . Neither the total Botox dose nor Botox into particular muscles differed between those with dysphagia and those without.

ASAIO J, 1992 Jul-Sep, 38(3), M248 - 52
Selective feedback control of spastic musculature in a canine model; Broniatowski M et al.; Muscular spasm, the etiology of which remains elusive, has been difficult to control . The authors, therefore, proposed selective restraint of forced contractions by a closed-loop circuit . In four dogs, the strap (n = 6) and thyroarytenoid (n = 4) muscles were submitted to tetanic contractions via bipolar supramaximal stimulation (30 Hz, 6 mA, 0.5 msec) of ansa hypoglossi and recurrent laryngeal nerves, respectively . Subsequent reduction of distance between two sonomicrometer crystals embedded into muscle was used in lieu of "spastic information" for a stimulator to deliver blocking signals through tripolar electrodes passed downstream . By modifying frequency (30-110 Hz) and current (60-95%) distribution between the central cathode and the peripheral anodes, significant relaxation (up to 100%) was recorded within "blocking windows," varying with each nerve (3-18 mA sweeps) . The selective restraint of unwanted contractions, leaving "normal" subthreshold tone undisturbed, may offer a more mature approach to spastic disorders than destructive procedures, such as nerve section and its chemical counterpart, botulinum toxin injection.

EMBO J, 1992 Jul, 11(7), 2465 - 71
Different types of K+ channel current are generated by different levels of a single mRNA; Honore E et al.; A cloned human voltage-sensitive K+ channel HLK3 which is present in T-lymphocytes and in the brain was expressed in Xenopus oocytes and after permanent transfection of a human B-lymphocyte cell line (IM9) . Injections of low cRNA concentrations into Xenopus oocytes led to the expression of a transient K+ current, with saturating current-voltage (I-V) relationship, which was abolished by repetitive stimulations due to a slow recovery from inactivation . This transient K+ channel current was fully inhibited by 10 nM charybdotoxin . Injection of high concentrations of the same RNA led to a non-inactivating K+ current, with linear I-V curve, which did not undergo use-dependent inactivation and was hardly sensitive to 10 nM charybdotoxin . Intermediate behaviour due to changing proportions of these two types of K+ channel expression were observed at intermediate RNA concentrations . Transient and non-inactivating K+ currents were also observed by both whole-cell and single channel patch-clamp recording from HLK3 transfected IM9 cells . The main conductance of the channel in the two different modes (inactivating and charybdotoxin-sensitive or non-inactivating and charybdotoxin-resistant) is the same (12-14 pS) . Destruction of the cytoskeletal elements with cytochalasin D, colchicine or botulinum C2 toxin in oocyte experiments prevented expression of the sustained mode of the K+ channel . The results suggest that the sustained mode obtained at high RNA concentrations corresponds to channel clustering involving cytoskeletal elements . This differential functional expression of K+ channels associated with different levels of mRNA appears as a new important factor to explain the biophysical and pharmacological diversity of voltage-sensitive K+ channels.

Schweiz Rundsch Med Prax, 1992 Jun 16, 81(25), 819 - 23
{The treatment of hemifacial spasm using botulinum toxin}; Schnyder H et al.; Hemifacial spasm is a disagreeable disturbance with involuntary unilateral twitching of the facial muscles . Its most common form is supposedly due to an irritation of the facial nerve at its proximal intracranial segment by vascular structures . Various forms of treatment including surgical procedures are employed, very often without satisfactory results but frequently involving the risk for severe complications . For a few years a new method has been using injection of botulinum toxin into the affected muscles, which in the majority of patients relieves the abnormal movements for about half a year; therefore, this very effective and secure procedure is recommended as first-line treatment of hemifacial spasm.

Curr Opin Neurol Neurosurg, 1992 Jun, 5(3), 301 - 7
The dystonias; Markham CH; The various dystonias have been found in at least five different hereditary backgrounds . The gene responsible for one of the dystonias, idiopathic torsion dystonia (ITD), lies on chromosome 9q32-34, with flanking markers now 1-2 cM apart . Magnetic resonance imaging and computerized tomography (CT) abnormalities in the basal ganglia, especially in the putamen, are found in many secondary dystonias . Botulinum toxin therapy is proving very useful in the treatment of focal dystonias.

Chin Med J (Engl), 1992 Jun, 105(6), 476 - 80
Treatment of blepharospasm, hemifacial spasm and strabismus with botulinum a toxin; Dai Z et al.; Thirty patients with blepharospasm, hemifacial spasm, strabismus and entropion were treated with botulinum A toxin giving satisfactory results . Rapid spasm relief, correction of strabismus and entropion were obtained . Only mild, transient and local side-effects occurred . The patients were followed up for 4-12 weeks with no recurrence . The clinical results show that local injection of a minute dose of botulinum A toxin in treating blepharospasm, hemifacial spasm, strabismus and entropion is a safe, effective and simple method of nonsurgical therapy.

Rinsho Shinkeigaku, 1992 Jun, 32(6), 637 - 8
{Botulinum toxin trial for spasticity}; Mezaki T et al.; We treated three patients with spastic paraparesis with botulinum toxin (BTX) . Two of them had scissors gait reflecting the hypertonus of adductors . In both, an injection of BTX to the responsible muscles improved gait, albeit transiently . The effect was shortlived, probably because increased tonus in non-injected portions compensated for the weakness of injected portions . In the third patient with no abnormality in tonus of the adductors, BTX injection in quadriceps caused a weakness with exacerbation of gait disturbance . We concluded that, in selected cases, BTX therapy is useful for spastic paraparesis to alleviate hypertonicity of the adductors.

J Protein Chem, 1992 Jun, 11(3), 255 - 64
Pepsin fragmentation of botulinum type E neurotoxin: isolation and characterization of 112, 48, 46, and 16 kD fragments; Gimenez JA et al.; Controlled digestion of approximately 150 kD single chain botulinum type E neurotoxin with pepsin at pH 6.0 produced 112, 48, 46, and 16 kD fragments . These were chromatographically purified; their locations in the approximately 1300 amino acid residue long neurotoxin were determined by identifying the amino terminal 10 residues of 112 and 48 kD fragments, 50 residues of 46 kD fragment, and 59 residues of 16 kD fragment . The 48 and 112 kD fragments contain the N-terminal segment of the neurotoxin (i.e., residue no . 1 to approximately 425 and 1 to approximately 990, respectively), the 46 kD fragment corresponds to approximately 407 residues of the C-terminal region, and the 16 kD fragment contains the approximately 140 residues from a segment nearer to the C-terminus . The 48 kD fragment is similar to the approximately 50 kD N-terminal light chain of the approximately 150 kD dichain neurotoxin, which is generated by tryptic cleavage of the approximately 150 kD single chain neurotoxin, and is separated from the approximately 100 kD C-terminal heavy chain by dithiothreitol (DTT) reduction of an intrachain disulfide bond in the presence of 2 M urea (Sathyamoorthy and DasGupta, J . Biol . Chem . 260, 10461, 1985) . The pepsin-generated 48 kD fragment, unlike the light chain, was isolated without exposure to DTT and urea . The single chain 112 kD fragment following trypsin digestion yielded 48 and 60 kD fragments that were separable after DTT reduction of the intrachain disulfide which links them . The N-terminal residues of the smaller fragment were identical to that of the single chain 150 kD neurotoxin; the single chain 112 kD fragment is therefore the neurotoxin minus the approximately 50 kD C-terminal half of the heavy chain . The biological activities of the 48 and 112 kD fragments can be demonstrated in permeabilized PC12 cells (Lomneth et al., J . Neurochem . 57, 1413, 1991); they inhibit norepinephrine release.

J Clin Neuroophthalmol, 1992 Jun, 12(2), 121 - 7
Effects of repeated botulinum toxin injections on orbicularis oculi muscle; Borodic GE et al.; Histologic evaluation was conducted on 12 orbicularis oculi specimens from 11 patients with essential blepharospasm and Meige's disease who had received an average of 11.3 injections of botulinum A toxin over 3.5 years . Denervation was demonstrated by the spread of acetylcholinesterase staining on muscle fibers when specimens were evaluated within 11 weeks of the last injection . When specimens were taken after 12 weeks, spread of acetylcholinesterase was confined to the neuromuscular junctions, with little fiber size variability resembling normal muscle . Fibrosis seen in three specimens could be correlated to prior surgery . Repeated injections of botulinum toxin into human muscle do not appear to cause irreversible muscle atrophy or other degenerative changes . Denervation changes (fiber size variability, acetylcholinesterase spread) appear to correlate to the time interval since the last injection.

Neurosci Lett, 1992 May 25, 139(2), 289 - 92
Differences in the temperature dependencies of uptake of botulinum and tetanus toxins in Aplysia neurons; Poulain B et al.; The respective neuroselective actions of botulinum type A (BoNT) and tetanus (TeTx) neurotoxins on cholinergic and non-cholinergic synapses of Aplysia are mainly due to differences in their extracellular neuronal targetting . Further information was gained on this neuroselectivity by examining the temperature dependencies of binding, internalization and intracellular action of both toxins . After reduction of temperature from 22 degrees C to 10 degrees C, the binding of neither BoNT nor TeTx was significantly altered whereas the neuronal uptake of BoNT, but not of TeTx, was prevented . Although TeTx internalization could be detected at the low temperature, its intracellular activity was greatly attenuated compared to that of BoNT . It is inferred that the uptake mechanisms are different for these two related but distinct toxins.

Masui, 1992 May, 41(5), 811 - 6
{Effect of botulinum A toxin injection in the management of hemifacial spasm}; Hakuto T et al.; The present study was carried out on the effect of botulinum A toxin, made in Japan, in the management of 30 hemifacial spasm patients . In almost all cases, alleviation of hemifacial spasm reached its peak in a few days . Availability was 93.3% and its effect lasted about 4 months . Side effects were observed in 12 patients (40%), but no serious complications were observed.

Postgrad Med, 1992 May 1, 91(6), 327 - 32, 334
Botulinum toxin therapy for neurologic disorders; Tim R et al.; In the last 20 years, the therapeutic uses of botulinum toxin, a potent neurotoxin, have been investigated . The agent produces chemical denervation of muscle, thereby causing atrophy and weakness . Studies have shown that injection of this agent is an effective therapy for focal dystonias, particularly blepharospasm, hemifacial spasm, and torticollis . Investigation continues into the role of botulinum toxin in the treatment of anismus, detrusor-sphincter dyssynergia, writers' cramp, and other disorders in which focal weakening of selected muscles could be useful.

Rinsho Shinkeigaku, 1992 May, 32(5), 527 - 31
{A case of bilateral blepharospasm responsive to edrophonium}; Funakawa I et al.; A case of bilateral blepharospasm who registered the efficacy of edrophonium was reported . The case is a 49-year-old female . She had been in good health until January, 1991 when she complained of difficulty in opening her eyes while driving . Thereafter the condition progressed to such a degree that she was unable to experience a comfortable life . Her blinking rate did not changed . The symptoms were triggered by stress or some physical action, such as walking or driving . They were attenuated by taking a bath, sleep or sedation . The severity of the symptoms varied during the day and from day to day . Neurological examination revealed bilateral spasms of the orbicular oculi muscles, and occasionally of the orbicular oris muscles, sternocleidmastoid muscles and the perinasal regions . Neither orolingual dyskinesia nor other involuntary movements were detected . Surface electromyography (EMG) disclosed tonic discharges mainly from the orbicular oculi muscles . The abnormal spasm disappeared with the injection of edrophonium chloride . The test for the serum antiacetylcholine receptor antibody was negative and a repetitive stimulation EMG showed no waning phenomenon . No thymoma or thymus abnormalities were detected by pneumomediastinography . A needle EMG revealed neurogenic change in the distal portion of the limbs . A single fiber EMG showed elongation of the jitter value and the blocking phenomenon . Although distigmine bromide was ineffective against the spasm, pyridostigmine bromide and the local injection of botulinum toxin were very effective.(ABSTRACT TRUNCATED AT 250 WORDS)

Aust N Z J Ophthalmol, 1992 May, 20(2), 121 - 7
The role of botulinum toxin in third nerve palsy; Saad N et al.; Isolated third nerve paresis is a rare diagnosis among patients presenting to the Botulinum Toxin Clinic at Moorfields Eye Hospital . Ten patients with this diagnosis are reviewed in this study . Head trauma is a common cause of third nerve palsy and is often severe enough to cause damage to fusion potential . If fusion is present and there is adequate adduction of the divergent eye, then botulinum toxin injection of the lateral rectus may induce long-term control of the ocular deviation . Three of the four patients who experienced long-term control of their ocular deviation following toxin injection shared these features . Toxin injected into the lateral rectus did not, however, reliably assess medial rectus function and therefore predict the outcome of horizontal squint surgery . Reasons for this are discussed.

Biochem J, 1992 May 1, 283 ( Pt 3), 899 - 904
Amylase release from streptolysin O-permeabilized pancreatic acinar cells . Effects of Ca2+, guanosine 5'-{gamma-thio}triphosphate, cyclic AMP, tetanus toxin and botulinum A toxin; Stecher B et al.; The molecular requirements for amylase release and the intracellular effects of botulinum A toxin and tetanus toxin on amylase release were investigated using rat pancreatic acinar cells permeabilized with streptolysin O . Micromolar concentrations of free Ca2+ evoked amylase release from these cells . Maximal release was observed in the presence of 30 microM free Ca2+ . Ca(2+)-stimulated, but not basal, amylase release was enhanced by guanosine 5'-{gamma-thio}triphosphate (GTP{S}) (3-4 fold) or cyclic AMP (1.5-2 fold) . Neither the two-chain forms of botulinum A toxin and tetanus toxin, under reducing conditions, nor the light chains of tetanus toxin, inhibited amylase release triggered by Ca2+, or combinations of Ca2+ + GTP{S} or Ca2+ + cAMP . The lack of inhibition was not due to inactivation of botulinum A toxin or tetanus toxin by pancreatic acinar cell proteolytic enzymes, as toxins previously incubated with permeabilized pancreatic acinar cells inhibited Ca(2+)-stimulated {3H}noradrenaline release from streptolysin O-permeabilized adrenal chromaffin cells . These data imply that clostridial neurotoxins inhibit a Ca(2+)-dependent mechanism which promotes exocytosis in neural and endocrine cells, but not in exocrine cells.

Blood, 1992 Apr 15, 79(8), 2056 - 67
Thrombin stimulation of human endothelial cell phospholipase D activity . Regulation by phospholipase C, protein kinase C, and cyclic adenosine 3'5'-monophosphate; Garcia JG et al.; The activation of membrane-bound phospholipase D (PLD) resulting in the generation of phosphatidic acid (PA) is increasingly recognized as an integral event in the initiation of a variety of cellular responses . We explored whether alpha-thrombin is a physiologic agonist for PLD activation in human umbilical vein endothelial cells (HUVEC) . HUVEC monolayers were labeled with {32Pi} and PLD activity determined by formation of the PLD metabolite {32P} phosphatidylethanol (PEt) in the presence of 5 g/L ethanol by thin-layer chromatography . alpha-Thrombin rapidly (1 minute) increased PA and PEt formation in a dose-dependent manner (10(-6) to 10(-10)) with maximal PLD stimulation achieved with 10 nmol/L alpha-thrombin producing a threefold to fourfold increase in PA and a sixfold to eightfold increase in PEt over controls at 15 minutes . Esterolytically active zeta-thrombin (10 nmol/L) and gamma-thrombin (1 mumol/L), but not inactive DIP-alpha-thrombin (1 mumol/L) also increased PLD activity . The role of Ca2+ flux in human endothelial cell PLD activation was investigated and PEt formation was significantly enhanced by Ca2+ ionophores A23187 and ionomycin (1 mumol/L, three-fold to fourfold increase in PEt) . Alpha-Thrombin-stimulated PEt formation was abolished (greater than 90% inhibition) with chelation of intracellular calcium (Ca2+i) by pretreatment with BAPTA-AM (25 mumol/L, 30 minutes) but only mildly attenuated (30% inhibition) by removal of extracellular calcium (Ca2+E) with EGTA (5 mmol/L) . The protein kinase C (PKC) inhibitor staurosporine reduced alpha-thrombin-induced PEt formation in a dose-dependent manner (10 mumol/L, 78% inhibition) and PKC downregulation with chronic PMA treatment (18 hours) also resulted in marked inhibition of alpha-thrombin-induced PEt formation . Neither pertussis nor botulinum C bacterial toxins significantly altered alpha-thrombin-induced PLD responses . In contrast, similar pretreatment with cholera toxin (1 microgram/mL, 60 minutes) consistently augmented alpha-thrombin-stimulated PLD activity by 50% to 90% . Comparable results were observed with agents which increased cAMP such as forskolin, 8-bromo cAMP, or dibutyryl cAMP and cholera toxin augmentation was abolished by 2-dideoxyadenosine, a competitive inhibitor of adenylyl cyclase activity . These studies demonstrate that alpha-thrombin is a potent stimulus for human PLD-mediated PA formation and that cyclic adenosine nucleotides modulate agonist-induced cellular PLD activity . In this model of PLD activation, alpha-thrombin receptor occupancy leads to the breakdown of phosphatidylinositol 4,5-bisphosphate catalyzed by phospholipase C producing the Ca2+ secretagogue IP3 and DAG.(ABSTRACT TRUNCATED AT 400 WORDS)

Cas Lek Cesk, 1992 Apr 10, 131(7), 213 - 6
{Botulinum toxin in the treatment of blepharospasm . Initial experience}; Ruzicka E et al.; The clinical effects of botulinum toxin A were evaluated in eight patients with idiopathic blepharospasm and hemifacial spasm . A substantial reduction or disappearance of muscle spasms were observed after the treatment . The action of botulinum toxin became noticeable 2 days after administration with a peak effect at 2 weeks . This effect diminished with muscle spasms returning about 12 weeks after administration . No serious adverse events were noted . The treatment with botulinum toxin has become method of choice in blepharospasm, it is proposed for the management of local muscle spasms of various origin.

Neurology, 1992 Apr, 42(4), 878 - 82
Botulinum toxin injection for spasmodic torticollis: increased magnitude of benefit with electromyographic assistance; Comella CL et al.; To determine the usefulness of EMG-assisted botulinum toxin (BOTOX) injections for the treatment of spasmodic torticollis (ST), we randomized 52 ST patients into two groups and studied them prospectively . In one group {(E+C)RX, N = 28}, the muscles were selected for BOTOX injection using both clinical and EMG examination and then injected with EMG assistance . In the second group {(C)RX, N = 24} the muscles were selected for BOTOX injection based solely on clinical examination and injected without EMG assistance . The percentage of patients showing any improvement after BOTOX as similar in both the (E+C)RX and (C)RX groups . A significantly greater magnitude of improvement was present in the (E+C)RX group, as well as a significantly greater number of patients with marked improvement . In particular, patients with retrocollis, head tilt, and shoulder elevation demonstrated additional benefit with EMG-assisted BOTOX injection . EMG assistance may be effective because the technique increases the ability to effectively identify and treat the deep cervical muscles.

Laryngoscope, 1992 Apr, 102(4), 400 - 6
Effects of botulinum toxin therapy in patients with adductor spasmodic dysphonia: acoustic, aerodynamic, and videoendoscopic findings; Zwirner P et al.; Botulinum toxin has been previously reported to be successful in the treatment of spasmodic dysphonia . To objectively document results, 11 patients with adductor spasmodic dysphonia who received unilateral treatment of the thyroarytenoid muscle were studied . Acoustic analyses and airflow rates during sustained phonation and flexible videoendoscopy were performed prior to, 1 week and 1 month after injection . 1 . Acoustic parameters demonstrated significant voice improvement, although abnormal characteristics remained . 2 . Mean airflow rates were increased 1 week after injection with almost normal values 1 month later . 3 . Videolaryngoscopy showed an effective reduction of intrinsic laryngeal muscle hyperfunction with less effect on extrinsic muscle activity . Interrelations between videolaryngoscopic rating scores, acoustic results and aerodynamic results are discussed.

J Physiol, 1992 Apr, 449, 479 - 92
Neural regulation of acetylcholine receptors in rat neonatal muscle; Bambrick LL et al.; 1 . The neuronal regulation of the developmental decline in skeletal muscle acetylcholine (ACh) receptors was studied by comparing the effects of sciatic nerve section or of neuromuscular blockade with botulinum toxin (BoTX) on this decline in neonatal and adult rats, using 125I-alpha-bungarotoxin (125I-BTX) as a ligand for the receptor alpha-subunit . 2 . The decline in 125I-BTX binding site concentration in neonatal rat triceps surae muscle homogenates towards low, adult levels followed a simple exponential with a time constant of 8 days . This decline occurred while the muscle is still rapidly growing, before the postnatal increase in numbers of sodium channels . It also preceded the decline in muscle ACh receptor alpha-subunit mRNA, reported in other studies, suggesting that subunit levels are not regulated only by mRNA availability . 3 . Muscle denervation in the first two weeks of life prevented this developmental decline . Denervation increased the concentration of 125I-BTX binding sites but the magnitude of this increase became progressively smaller as the muscle matured, showing that removal of innervation during adult life does not revert the muscle, in toto, to its pre-innervation state . 4 . Blockade of neuromuscular activity with BoTX increased 125I-BTX binding sites to a lesser extent than muscle denervation during neonatal life . This lesser effect of BoTX blockade contrasts with the equal effects of BoTX blockade and denervation in the adult.

J Med Assoc Thai, 1992 Apr, 75(4), 199 - 203
Two hundred and fifty patients with hemifacial spasm treated with botulinum toxin injection; Poungvarin N et al.; Two hundred and fifty patients with hemifacial spasm from the Movement Disorder Clinic, at Siriraj Hospital have been treated with botulinum toxin injection since January 1989 as a collaborating research project with Smith-Kettlewell Eye Research Institute in San Francisco . Each patient received 30 units in four injection sites over the hyperkinetic facial muscles . There were 169 female and 81 male patients, the sex ratio of female to male was 2.1:1 . The mean age of all patients was 50.2 +/- 12.6 years with the range of 22 to 78 years . The majority of patients had been suffering for 3-10 years . The results of botulinum toxin injection were classified as excellent in 81.2 per cent, moderate improvement 10.0 per cent, mild improvement 6.8 per cent and no improvement or worse in 2.0 per cent . There were complications of mild transient facial weakness in 44 patients (17.6%) mild ptosis in 7 patients (2.8%) and excessive lacrimation in 1 patient (0.4%) . The effect of botulinum toxin treatment lasted for 3-6 months duration . Botulinum toxin injection is a simple and effective out-patient treatment for patients with hemifacial spasm with no systemic side effects and minor transient local complications.

Isr J Med Sci, 1992 Mar-Apr, 28(3-4), 183 - 5
Surgery of the internal acoustic meatus and the cerebello-pontine angle; Bebear JP et al.; Tumoral and functional surgery of the cerebellopontine angle and the internal acoustic meatus has been performed in our department for more than 20 years . Acoustic neurinomas (700 cases operated to date) represent the great bulk of this surgery . Since 1985, we have drastically modified our approach to these tumors (220 cases) . Large tumors are now dealt with, regardless of the patient's hearing, through a widened translabyrinthine approach . Small tumors with normal hearing are operated on through a suboccipital or a retrolabyrinthine approach depending on the tumoral extension in the internal acoustic meatus . The facial nerve function was preserved in 83% of the cases . Hearing was saved only 6 times in 45 attempts, despite 23 cases of apparent anatomical conservation of the cochlear nerve . Vestibular neurectomy, carried out through a middle cranial fossa or through a retrolabyrinthine approach, ameliorated vertigo in 95% of cases . The major operative risks are regressive facial paresis and secondary total deafness (10% of cases) . Surgical treatment of facial hemispasm by neurovascular decompression is very effective, but not without risk . We now prefer to use botulin toxin therapy.

Microbiol Rev, 1992 Mar, 56(1), 80 - 99
Properties and use of botulinum toxin and other microbial neurotoxins in medicine; Schantz EJ et al.; Crystalline botulinum toxin type A was licensed in December 1989 by the Food and Drug Administration for treatment of certain spasmodic muscle disorders following 10 or more years of experimental treatment on human volunteers . Botulinum toxin exerts its action on a muscle indirectly by blocking the release of the neurotransmitter acetylcholine at the nerve ending, resulting in reduced muscle activity or paralysis . The injection of only nanogram quantities (1 ng = 30 mouse 50% lethal doses {U}) of the toxin into a spastic muscle is required to bring about the desired muscle control . The type A toxin produced in anaerobic culture and purified in crystalline form has a specific toxicity in mice of 3 x 10(7) U/mg . The crystalline toxin is a high-molecular-weight protein of 900,000 Mr and is composed of two molecules of neurotoxin (ca . 150,000 Mr) noncovalently bound to nontoxic proteins that play an important role in the stability of the toxic unit and its effective toxicity . Because the toxin is administered by injection directly into neuromuscular tissue, the methods of culturing and purification are vital . Its chemical, physical, and biological properties as applied to its use in medicine are described . Dilution and drying of the toxin for dispensing causes some detoxification, and the mouse assay is the only means of evaluation for human treatment . Other microbial neurotoxins may have uses in medicine; these include serotypes of botulinum toxins and tetanus toxin . Certain neurotoxins produced by dinoflagellates, including saxitoxin and tetrodotoxin, cause muscle paralysis through their effect on the action potential at the voltage-gated sodium channel . Saxitoxin used with anaesthetics lengthens the effect of the anaesthetic and may enhance the effectiveness of other medical drugs . Combining toxins with drugs could increase their effectiveness in treatment of human disease.

J Neurol Neurosurg Psychiatry, 1992 Mar, 55(3), 229 - 31
Psychological functioning before and after treatment of torticollis with botulinum toxin; Jahanshahi M et al.; The impact of botulinum toxin injection on psychological function was assessed in 26 patients with idiopathic torticollis . Eighty five per cent of the patients and 88% of the relatives considered torticollis to be better following the injections . Symptomatic improvement with the injections was associated with significant reduction of depression and disability, but non-significant improvement in body concept, and self-esteem . This concordant pattern of change in symptoms and psychological function with the injections supports the proposal that in torticollis depression and disability are consequences of the postural abnormality of the head.

Neurology, 1992 Mar, 42(3 Pt 1), 627 - 30
Botulinum toxin therapy for limb dystonias; Yoshimura DM et al.; We investigated the effectiveness of botulinum toxin in 17 patients with limb dystonias (10 with occupational cramps, three with idiopathic dystonia unrelated to activity, and two each with post-stroke and parkinsonian dystonia) in a placebo-controlled, blinded study . We identified affected muscles clinically and by recording the EMG from implanted wire electrodes at rest and during performance of tasks that precipitated abnormal postures . There were three injections given with graded doses of toxin (average doses, 5 to 10, 10 to 20, and 20 to 40 units per muscle) and one with placebo, in random order . Subjective improvement occurred after 53% of injections of botulinum toxin, and this was substantial in 24% . Only one patient (7%) improved after placebo injection . Subjective improvement occurred in 82% of patients with at least one dose of toxin, lasting for 1 to 4 months . Response rates were similar between clinical groups . Objective evaluation failed to demonstrate significant improvement following treatment with toxin compared with placebo . The major side effect was transient focal weakness after 53% of injections of toxin.

Neurology, 1992 Mar, 42(3 Pt 1), 602 - 6
Persistent unilateral tibialis anterior muscle hypertrophy with complex repetitive discharges and myalgia: report of two unique cases and response to botulinum toxin; Nix WA et al.; Unilateral enlargement of the tibialis anterior muscle associated with complex repetitive discharges occurred over several months in two patients and was preceded by pain and numbness in the lower leg . Neuroradiologic investigations excluded a compressive radiculopathy, but pharmacologic and neurophysiologic studies suggested a neurogenic basis for the muscle hypertrophy . Botulinum toxin A injection into the hypertrophied muscles led to a decreased muscle volume and cessation of muscle pain.

Am J Respir Cell Mol Biol, 1992 Mar, 6(3), 253 - 9
Identification of ras and ras-related low-molecular-mass GTP-binding proteins associated with rat lung lamellar bodies; Rubins JB et al.; Recent evidence from genetic experiments in yeast and from studies using guanosine triphosphate (GTP) analogues in mammalian cells suggests a key role for low-molecular-mass GTP-binding proteins (LMM-GBPs) (Mr 19 to 28 kD) in processes of intracellular vesicular sorting and secretion . Assembly and exocytosis of the lamellar body (LB), the secretory organelle of the pulmonary alveolar type 2 pneumocyte, may be regulated by LMM-GBPs . We used {alpha-32P}GTP binding to Western blotted proteins, ultraviolet crosslinking of {alpha-32P}GTP to membrane proteins, immunoblotting with specific antisera, and botulinum exoenzyme C3-catalyzed ADP ribosylation to detect LMM-GBPs in LB . With the first two techniques, we have identified six LMM-GBPs of approximately 27, 25.5, 24.5, 23, 22, and 21 kD that are enriched in a highly purified LB fraction compared with type 2 pneumocyte homogenate, crude membranes, and cytosol . Further characterization of the LB LMM-GBPs by immunoblotting revealed that ras p21 is greatly enriched in the LB fraction compared with other type 2 pneumocyte fractions . In addition, botulinum exoenzyme C3 catalyzed the ADP ribosylation of 20- to 21-kD proteins that were similarly enriched in the LB fraction . In contrast, a monospecific antibody to ADP-ribosylation factor reacted with a 19-kD protein only in the type 2 pneumocyte homogenate and cytosol fractions . Monospecific antibodies to yeast Sec4 protein and to rab 3A did not react with any type 2 pneumocyte proteins . The LMM-GBPs specifically associated with LB may participate in intracellular events required for surfactant packaging and secretion.

FEBS Lett, 1992 Feb 24, 298(2-3), 118 - 22
The effect of botulinum toxin type D on the triggered and constitutive exocytosis/endocytosis cycles in cultures of bovine adrenal medullary cells; von Grafenstein H et al.; The extracellular fluid phase marker, horseradish peroxidase, enters chromaffin cells when triggered to secrete catecholamine . This triggered uptake, like secretion, is abolished in cells pre-incubated with botulinum toxin . Endocytosis of horseradish peroxidase into unstimulated cells is unaffected by botulinum toxin but is inhibited when the temperature is reduced . Once internalised by the unstimulated cells, horseradish peroxidase is released back into the extracellular fluid, the rate of release being temperature sensitive but unaffected by carbamylcholine or botulinum toxin . These results suggest that triggered exocytosis is a necessary event to precede triggered endocytosis, and that botulinum toxin may affect only the triggered exocytosis/endocytosis cycle and not the constitutive cycle.

Lancet, 1992 Feb 22, 339(8791), 457 - 8
Stridor and focal laryngeal dystonia; Marion MH et al.; Fibreoptic laryngoscopy in 6 patients with laryngeal stridor showed immobile vocal cords in a paramedian position but no other local cause . Thus a diagnosis of Gerhardt's syndrome, usually ascribed to paralysis of vocal-cord abductor muscles, was made in 3 patients who had no other signs or symptoms of dystonia, and in 3 patients who had multifocal dystonia . Electromyography (EMG) showed evidence of overactivity of vocal-cord adductors, with no evidence of denervation in the abductor muscles . Botulinum toxin injection of the overactive thyroarytenoid muscles abolished stridor . These clinical and EMG findings indicate that Gerhardt's syndrome is not caused by paralysis of vocal-cord abductors, but represents a focal laryngeal dystonia which may be treatable by botulinum toxin injections of vocal-cord adductor muscles rather than by arytenoidopexy or tracheostomy.

Laryngoscope, 1992 Feb, 102(2), 163 - 7
Abductor laryngeal dystonia: a series treated with botulinum toxin; Blitzer A et al.; Abductor laryngeal dystonia (LD) is characterized by a hoarse voice quality which is broken up by breathy or whispered portions . Botulinum toxin injection (Botox) has been a safe and effective treatment for adductor laryngeal dystonia and is currently accepted medical therapy . As an extension of the established treatment program, in 1989 treatment of abductor LD was initiated . Thirty-two patients have been treated by sequential percutaneous electromyogram-guided (EMG) injections of the posterior cricoarytenoid (PCA) muscles . Most patients required treatment of both PCA muscles and improved to an average of 70% of normal voice . Patients who had a preexisting tremor, evidence of dystonia in other muscle groups, vocal tremor, or respiratory dysrhythmia had less improvement . Ten patients also required injection of the cricothyroid muscles and/or type I laryngoplasty.

Aust N Z J Ophthalmol, 1992 Feb, 20(1), 41 - 6
Modern management of sixth nerve palsy; Lee J; Between November 1982 and September 1991, 179 patients with unilateral or bilateral sixth nerve palsy were treated in the Botulinum Toxin Strabismus Clinic at Moorfields Eye Hospital . Indications for treatment included prophylaxis (as part of a prospective treatment trial), maintenance therapy, diagnosis and adjunct to surgical therapy . A management plan for established sixth nerve palsy based on the rational use of toxin and surgery is suggested.

J Neurosci Res, 1992 Feb, 31(2), 318 - 26
Miniature endplate potentials induced by ammonium chloride, hypertonic shock, and botulinum toxin; Vautrin J; Intracellular recordings were made at the neuromuscular junction (NMJ) of the mouse diaphragm to study alteration of miniature endplate potential (MEPP) amplitude and rise time after different treatments . Following either hyperosmotic shock or 3 to 5 min of incubation in 10 to 50 mM ammonium chloride (NH4Cl) (replacing NaCl, a treatment which is known to raise intracellular pH) MEPP frequencies increased and the amplitudes of MEPPs decreased . These treatments as well as type A botulinum toxin (BoTx) gradually prolonged the rising phase of some MEPPs, which increased their time-to-peak (slow-MEPPs; Vautrin and Kriebel: Neuroscience 41:71-88, 1991) and increased eventually their amplitude . Fasciculation after hyperosmotic shock or during NH4Cl challenge was blocked by D-tubocurarine and was due to large slow-MEPPs that reached threshold for the muscle fiber action potential . The development of fasciculation provided the time course for the development of giant-MEPPs . Increased frequency of giant MEPP is accompanied by a block of the nerve-evoked muscle contraction . Effects of BoTx on spontaneous release were functionally antagonized either by NH4Cl or hyperosmotic shock . NH4Cl delayed BoTx blockage of bell-MEPPs . Data suggest that BoTx alters the formation of transmitter packets gradually but similarly to other treatments which increase incidence of skew-MEPPs.

Naunyn Schmiedebergs Arch Pharmacol, 1992 Feb, 345(2), 227 - 34
Reductive cleavage of tetanus toxin and botulinum neurotoxin A by the thioredoxin system from brain . Evidence for two redox isomers of tetanus toxin; Kistner A et al.; Inhibition of neurotransmitter release by tetanus toxin and botulinum neurotoxin A can be mimicked by intracellular application of the corresponding toxin light chains . The aim of this study was to determine whether the two-chain toxins are reduced by brain preparations to yield free light chains which would represent the ultimate toxins . The interchain disulfide of two-chain tetanus toxin was cleaved by rat cortex homogenate fortified with NADPH . Reduction was promoted further by addition of thioredoxin . Thioredoxin reductase was demonstrated in and purified from porcine brain cortex . The thioredoxin system which consisted of purified enzyme, thioredoxin and NADPH reduced both toxins . The resulting light chains appeared homogeneous in SDS gel electrophoresis . The complementary heavy chain of tetanus but not of botulinum toxin migrated in two bands, the faster one with the velocity of heavy chain obtained by chemical reduction . The major, slower form was converted into the faster by chemical but not by enzymatic reduction . Tetanus toxin, whether in its single-chain or two-chain version also occurred in two forms which differed by their electrophoretic mobility . The two forms of single-chain toxin were interconverted by chemical reduction or oxidation but not by the thioredoxin system . It is concluded that a) a thioredoxin system in brain tissue reduces the interchain disulfide of two-chain tetanus toxin and botulinum neurotoxin A, b) tetanus toxin but not botulinum neurotoxin A consists of two electrophoretically distinct forms which differ by the thiol-disulfide status of their heavy chains, c) the disulfide loop within the heavy chain of tetanus toxin is resistant to the thioredoxin system.

J Pharmacol Exp Ther, 1992 Feb, 260(2), 859 - 64
Botulinum C2 toxin and steroid production in adrenal Y-1 cells: the role of microfilaments in the toxin-induced increase in steroid release; Considine RV et al.; Exposure of adrenal Y-1 cells to C2 toxin results in an increase in steroid release that is accompanied by a rounding of the cell . The actions of C2 toxin mimic those of adrenocorticotropin and cholera toxin except that there is no increase in intracellular cyclic AMP content . In the present study we provide evidence that C2 toxin increases steroid output from Y-1 cells through an alteration in the microfilament network of the cell . C2 toxin significantly increased steroid output after 3 hr of exposure . This effect was accompanied by a significant increase in the transport of {3H}cholesterol to the mitochondrial fraction, independent of cholesterol uptake by the cell . The toxin was unable to increase steroid output from cells prerounded in suspension culture . The protease inhibitors benzamidine and phenylmethylsulfonyl fluoride did not attenuate the ability of C2 toxin to alter the morphology of Y-1 cells . A 3-hr exposure to C2 toxin resulted in the ADP-ribosylation of 50 to 60% of the total actin pool . Fluorescein isothiocyanate-labeled phalloidin visualization of the cytoskeleton of toxin-treated cells confirmed that the toxin caused a decrease in the stress fiber network . C2 toxin treatment of a protein kinase A mutant Y-1 cell (Kin 8) resulted in morphological changes and an increase in steroid output that was not different from that observed for wild type Y-1 cells . The data suggest that C2 toxin increases steroid output from adrenal Y-1 cells by a cyclic AMP-independent mechanism that involves the microfilament network of the cell.

J Dermatol Surg Oncol, 1992 Jan, 18(1), 17 - 21
Treatment of glabellar frown lines with C . botulinum-A exotoxin; Carruthers JD et al.; Eighteen patients with glabellar frown lines were treated with C . botulinum-A exotoxin . Sixteen of the 17 patients followed showed improvement for periods ranging from 3 months to 11 months . Side-effects were minimal and transient . Because C . botulinum-A exotoxin therapy of glabellar frown lines treats the underlying cause of these lines, it is more effective than soft tissue augmentation although this improvement is temporary . Treatment with C . botulinum-A exotoxin is a simple, safe procedure.

Head Neck, 1992 Jan-Feb, 14(1), 33 - 7
Botulinum a toxin for spasmodic torticollis: multiple vs single injection points per muscle; Borodic GE et al.; Eighty-six injections in 49 patients with adult onset spasmodic torticollis were evaluated for efficacy with respect to single point per muscle versus multiple point per muscle injection techniques . Parameters of the syndrome assessed were pain, posture deformity, range of cervical motion, disfigurement, cervical muscle hypertrophy, activity limitation, and degree of involuntary movement . The multiple point per muscle injection strategy appeared superior to the single injection per muscle technique with respect to pain (p less than 0.002, chi-square), posture deformity (p less than 0.001), range of motion (p less than 0.001), and improvement in activity endurance (p less than 0.001) . No significant differences were noted with respect to cervical muscle hypertrophy or degree of involuntary movements, although the injections were considered beneficial in both groups to these syndrome components.

Rev Neurol (Paris), 1992, 148(3), 212 - 4
{Treatment of spasticity with botulinum toxin}; Memin B et al.; Spasticity following upper motor neuron lesion can be alleviated by few treatments such as physiotherapy, drugs and neurosurgery . However, they all have side effects, limitations or lack of selectivity . We tentatively used the paralyzing effects of botulinum toxin . Since the late 1970's the use of this toxin has increased and it has been extended to numerous muscles and diseases of various causes . In this pilot and open study we use botulinum toxin in spasticity . Eight patients (7 stroke, 1 head injury) with longstanding severe spasticity (minimum: 12 months, maximum: 15 years) were included . Spasticity greatly interfered with their activity in daily life and was resistant to oral antispastic medications . Six patients suffered from pain and 4 had cutaneous lesions especially maceration of the palm of the hand . A-botulinum toxin was injected with a 30-gauged needle . The sites chosen for injection were the following muscles: biceps brachii, brachioradialis, flexor digitorum, flexor carpi, tibialis anterior, flexor digitorum longus . Altogether 41 injections were performed . There were no side effects . Spasticity was improved in all patients . Five patients reported significant pain relief on a visual analogical scale . Most of them reported a benefit in their limb tone and referred to subjective improvement in the activity of daily life and nursing . The beneficial effects of one injection lasted more than 5 months . Seven patients received a second course of treatment . A double-blind study of botulinum toxin in spasticity is to be undertaken to assess its effectiveness and safety when prescribed in the required dose to treat this condition.

Rev Neurol (Paris), 1992, 148(3), 180 - 3
{Spasmodic dysphonia . Investigation and therapeutic methods}; Marion MH et al.; We report the successful treatment of three patients with adduction spasmodic dysphonia by direct injection of botulinum toxin into the vocal cord(s) . This was achieved under electromyographic control, and this and other otolaryngeal techniques were used to monitor treatment and study this often puzzling condition.

Eur Neurol, 1992, 32(2), 112 - 7
Botulinum toxin as a treatment for blepharospasm, spasmodic torticollis and hemifacial spasm; Albanese A et al.; Fifty-two patients affected by focal dystonia or hemifacial spasm were treated with repeated injections of botulinum toxin . A clinical improvement was observed in all patients with blepharospasm; clinical benefit had a mean duration of 10 weeks . Clinical results were less impressive, but also favorable in patients affected by spasmodic torticollis and by hemifacial spasm . In the latter, the incidence of drug-induced paresis was much higher than that observed in patients with blepharospasm, even though the doses of toxin injected were significantly lower.

Acta Neurol Scand, 1992 Jan, 85(1), 55 - 7
Treatment of idiopathic hemifacial spasm with botulinum toxin; Yu YL et al.; Twelve patients with idiopathic hemifacial spasm received treatment with botulinum toxin A over a period of 18 months . Of 76 treatments given, most (94.7%) led to successful relief of eyelid spasms and all treatments were successful for perioral and lower facial muscle spasms . An average dose of 9.3 units of toxin per session was given to produce a mean interval of relief of 10.8 weeks . Blepharoptosis was the only ocular side effect; it was mild, reversible and occurred in 2 patients . However, lower facial palsy was frequent (9 patients); it was mild to moderate in severity but only partially reversible in 8 patients . Dosage for lower facial muscles should therefore be reduced.

J Neurol, 1992 Jan, 239(1), 9 - 15
Clinical and polymyographic investigation of spasmodic torticollis; Deuschl G et al.; Polymyographic recordings were used to identify the most dystonic muscles suitable for local injection with botulinum toxin in 100 patients with spasmodic torticollis (TS) . Rotating TS (72% of the patients) was due to dystonic activity of the splenius muscle ipsilateral to and/or the sternocleidomastoid muscle contralateral to the side of chin deviation . One-third of these patients had also dystonic activation of the contralateral splenius muscle and, rarely, the contralateral trapezius muscle . Ten patients had laterocollis due to dystonic activation of all recorded muscles on one side of the neck . Nine patients had retrocollis due to activity of both splenius muscles and rarely additional activity in both trapezius muscles . The type of dystonic muscle activity was found to be tonic, phasic or tremulous . Besides the evaluation of spontaneous dystonic EMG activity further examination during the "geste antagoniste" or the muscle activity during rotating head movements can provide additional information . It is concluded that polymyography may provide a rationale for identifying the dystonic muscles underlying the different forms of TS . It may prove to be helpful for the successful therapy with botulinum toxin and may be useful in differentiating tremulous torticollis from other types of head tremor.

J Neurol, 1992 Jan, 239(1), 5 - 8
The management of blepharospasm and hemifacial spasm; Elston JS; The aetiology of blepharospasm and hemifacial spasm is different, but both produce involuntary eye closure and facial movements which do not respond to systemic drug treatment . The introduction of therapeutic focal muscle weakening with botulinum toxin injections in the early 1980s appeared to offer great promise in the management of these conditions . In this paper the results of botulinum toxin treatment of 234 patients with blepharospasm and 73 patients with hemifacial spasm over a 7-year period have been analysed . Most patients receive sustained benefit from repeated injections whilst side-effects become less frequent . A clinically recognisable subgroup of patients with blepharospasm respond poorly and may be better treated surgically.

J Neurol, 1992 Jan, 239(1), 21 - 5
Treatment of spasmodic torticollis with local injections of botulinum toxin . One-year follow-up in 37 patients; Poewe W et al.; Thirty-seven patients with spasmodic torticollis (cervical dystonia) who received repeated local injections of botulinum toxin have been followed up for a mean period of 12.3 (10-29) months, during which time 138 treatment sessions were performed . Mean doses per muscle averaged 320 mouse units (mu; range 160-1000 mu botulinum toxin A prepared by CAMR, Porton Down, UK) . Eighty-six per cent of patients experienced significant improvement of posture and 84% of those with pain had relief following the first injection . Muscular patterns of recurrent torticollis were relatively constant and in most patients efficacy was maintained with subsequent injections, while 15% of all follow-up sessions failed . Only 2 of 37 patients were consistent nonresponders; 22% and 10% of all sessions were complicated by transient dysphagia and weakness of neck muscles, respectively . It is concluded that local botulinum toxin injections can be a safe and efficaceous long-term treatment of spasmodic torticollis and that optimal doses should be between 200 and 400 mu/muscle.

J Neurol, 1992 Jan, 239(1), 1 - 4
Treatment of cervical dystonia hand spasms and laryngeal dystonia with botulinum toxin; Lees AJ et al.; One hundred and twenty-six patients with different forms of focal dystonia (89 with cervical dystonia, 12 with hand cramps and 25 with laryngeal dystonia) were treated with localised injections of botulinum toxin . Mean doses per muscle were 200 mouse units (m.u.) for treating cervical dystonia, 40-120 m.u . for forearm muscles in writers' cramp and 3.7 m.u . for the thyroarytenoid muscle in laryngeal dystonia . Responder rates have been above 80% in all patient groups and beneficial effects could be reproduced over follow-up periods of up to 4 years . The commonest side-effects were dysphagia after treatment of spasmodic torticollis, weakness of neighbouring muscles after injections for hand cramps and breathiness and hypophonia following laryngeal injections . All these were transient and generally well tolerated . It is concluded that botulinum toxin injections are a safe and effective treatment in all three types of focal dystonia.

Mov Disord, 1992, 7(2), 171 - 7
Severe but temporary injury to rabbit orbicularis oculi muscle using dihematoporphyrin ether and laser photochemomyectomy; Wirtschafter JD et al.; The use of local dihematoporphyrin ether (DHE) injections, followed by laser light activation, was investigated as a potential permanent myectomy treatment for muscle spasms, in particular blepharospasm and hemifacial spasm . DHE was injected into the eyelids of rabbits, followed by laser activation, as used in photochemotherapy . Four days after treatment, histological examination indicated that doses of greater than or equal to 0.5 mg of DHE and laser treatment with an energy density of at least 100 J/cm2 resulted in an almost total destruction of the orbicularis oculi muscle in the treated eyelid . The amount of muscle injury was dependent on both dose of DHE and energy density levels . Histologically, the tarsal glands and conjunctiva were damaged . Glandular tissue was markedly reduced, and the conjunctival epithelium showed hyperplasia and a loss of mucous cells . Six months after DHE and laser treatment, the majority of the muscle tissue had regenerated, although there was evidence of previous injury . While DHE injections combined with laser light activation were lethal to muscle at the site of treatment, this treatment was not permanent . The orbicularis oculi muscle retained its ability to regenerate . However, photochemomyectomy may be studied further as an adjuvant treatment to temporarily injure and debulk large muscles when botulinum toxin is contraindicated due to the large doses involved or as a permanent treatment when used together with an antimitotic agent such as doxorubicin.

Ophthal Plast Reconstr Surg, 1992, 8(2), 137 - 42
Botulinum A toxin for (expressionistic) ptosis overcorrection after frontalis sling; Borodic GE; Botulinum A toxin was injected into the frontalis muscle in two patients with complete third nerve palsies to limit intermittent upper lid retraction after a frontalis sling procedure . This form of lid retraction is noted during periods of active facial movement with occipitofrontalis muscle contraction . Although upper lid position may be symmetric when the facial muscles are adynamic, the upper lid may retract during periods of active facial expression . This type of lid retraction was corrected using Botulinum A toxin injections into the frontalis muscles, without affecting the lid position when the facial muscles are adynamic . Both improvement in appearance and intermittent exposure were noted in both cases . Additionally, a blunting of the transverse forehead creases occurred over a defined area after this injection, representing a clinical example of a denervation field produced by a point injection of botulinum toxin.

Acta Neuropathol (Berl), 1992, 84(1), 89 - 93
The effect of nerve crush and botulinum toxin on lead uptake in motor axons; Pamphlett R et al.; After lead (Pb) is injected into striated muscle it binds to the sarcolemma of the neuromuscular junction (NMJ) and crosses into the terminal axons of motor neurons . To find out whether this intra-axonal accumulation of Pb is due to active transport or to diffusion down a concentration gradient, Pb uptake into motor axons of mice was studied at active and inactive NMJs . Twenty-four hours after sciatic nerve crush, 0.1 ml of 5% lead nitrate was injected into the tibialis anterior muscle and 30 min later the location of Pb was sought with electron microscopy and X-ray elemental analysis . A greatly reduced amount of Pb entered the axons after nerve crush compared to non-nerve crush animals, indicating that an active NMJ is required for intra-axonal Pb accumulation . To test if Pb could be entering the axon via recycling vesicles, botulinum toxin (BoTx) was injected into the muscle 24 h before Pb injection . There was no difference in intra-axonal Pb uptake in control and BoTx-injected animals, indicating that Pb is unlikely to use recycled vesicles to enter the axon.

Trans Am Ophthalmol Soc, 1992, 90, 361 - 7; discussion 367-71
Botulinum alignment for congenital esotropia; Ing MR; This is the first report of a group of patients with congenital esotropia examined for motor and sensory evidence of binocularity a minimum of 3 years after alignment by botulinum . Evidence for binocularity was clearly present in approximately one half of the patients . Lag time to satisfactory alignment was at least 1 month (average, 5 months) following the initial botulinum injection . The results must be considered preliminary . However, when these results are compared with those of patients with congenital esotropia aligned by incisional surgery by age 2 years and examined with the same testing devices by this same investigator, botulinum alignment appears to be less effective than surgical alignment in establishing evidence for binocularity (P < 0.005).

J Neural Transm Suppl, 1992, 38, 91 - 104
Dystonia--a clinical, neuropathological and therapeutic review; Wissel J et al.; Dystonia is a syndrome characterized by sustained muscle contractions frequently causing twisting and repetitive movements or abnormal posture . For diagnosis, prognosis and therapy, it is useful to classify dystonia with regard to types of abnormal movements present, their mode of activation and topographical distribution taking into account age of onset, and etiology . The majority of cases are idiopathic, or primary dystonias, while in a minority environmental, structural, or metabolic causes can be identified . Primary dystonias can be familial or sporadic . The most important neurophysiological phenomenon in dystonia is pathological cocontraction of antagonistic muscles, while there is no consistent neuropathological abnormality in idiopathic dystonia . Causal therapies for dystonia are only possible in a few symptomatic forms (M . Wilson, Segawa-syndrome) . As a rule, treatment has to be symptomatic but results of systemic pharmacotherapies remain disappointing . For adult onset focal dystonias, a breakthrough in symptomatic therapy has been achieved with local "chemical" denervation by means of botulinum toxin type A injections.

Eye, 1992, 6 ( Pt 4), 386 - 90
Management of VIth nerve palsy--avoiding unnecessary surgery; Riordan-Eva P et al.; Unresolved VIth nerve palsy that is not adequately controlled by an abnormal head posture or prisms can be very suitably treated by surgery . It is however essential to differentiate partially recovered palsies, which are amenable to horizontal rectus surgery, from unrecovered palsies, which must be treated initially by a vertical muscle transposition procedure . Botulinum toxin is a valuable tool in making this distinction . It also facilitates full tendon transposition in unrecovered palsies, which appears to produce the best functional outcome of all the transposition procedures, with a reduction in the need for further surgery . A study of the surgical management of 12 patients with partially recovered VIth nerve palsy and 59 patients with unrecovered palsy provides clear guidelines on how to attain a successful functional outcome with the minimum amount of surgery.

Bioseparation, 1992-93, 3(5), 267 - 83
Purification of bacterial exotoxins . The case of botulinum, tetanus, anthrax, pertussis and cholera toxins; Pasechnik VA et al.; Bacterial protein toxins and their fragments have been isolated and purified for various reasons, including the development of efficient vaccines and for methods of identification of bacterial agents causing disease . This activity continues today but a new area of bacterial protein toxin research has recently emerged . Since it was shown that toxin molecules comprise several types of biological activity within their structural domains, it was suggested to use these domains (and their combinations) as biochemical tools for developing novel agents for disease imaging and and/or relieving . In this way eukaryotic cell-receptor specific fusion toxins have been developed to prevent malignancy in human . While human clinical trials of these preparations have only recently begun, the preliminary clinical findings are promising . Also fusion proteins which combine independent immunodominant epitopes from different antigens have also been developed thus opening a way for the generation of new vaccines for both human and veterinary use . Receptor binding fragments of microbial toxins when combined with other molecules may be useful in delivering these molecules into the cell . In this way novel agents may be developed with a potential for inducing specific changes at the molecular level for the correction of metabolic disorders causing human and animal diseases . Bacterial protein toxins such as anthrax, botulinum, cholera, pertussis and tetanus for which considerable progress has been achieved in structure-function analysis are promising candidates for such research . Particularly exciting appears the idea of extending this research to the cells of the nervous system, exploiting the unique specificity of the botulinum or tetanus toxin fragments which may bring long desired methods for treatment of various disorders of the nervous system . Data on functional domains of these toxins as well as methods of purification of the whole toxins and their fragments are considered in this review as they form a base for their further structure-function analysis and engineering applications.

J Neural Transm Gen Sect, 1992, 90(2), 87 - 102
Binding of botulinum neurotoxin to pure cholinergic nerve terminals isolated from the electric organ of Torpedo; Blasi J et al.; Torpedo electric organ has been used to study the binding of botulinum neurotoxin type A to pure cholinergic synaptosomes and presynaptic plasma membrane . 125I-labeled botulinum neurotoxin type A exhibits specific binding to cholinergic fractions . Two binding sites have been determined according to data analysis: a high affinity binding site (synaptosomes: Kd = 0.11 +/- 0.03 nM, Bmax = 50 +/- 10 fmol.mg prot-1; presynaptic plasma membrane: Kd = 0.2 +/- 0.05 nM, Bmax = 150 +/- 15 fmol.mg prot-1) and a low affinity binding site (synaptosomes: Kd approximately 26 nM, Bmax approximately 7.5 pmol.mg prot-1; presynaptic plasma membrane: Kd approximately 30 nM, Bmax approximately 52 pmol.mg prot-1) . The binding of 125I-botulinum neurotoxin type A is decreased by previous treatment of synaptosomes by neuraminidase and trypsin, and by a preincubation with bovine brain gangliosides or antiserum raised against Torpedo presynaptic plasma membrane . When presynaptic plasma membranes are blotted to nitrocellulose sheet, either 125I-botulinum neurotoxin or botulinum toxin-gold complexes bind to a M(r) approximately 140,000 protein . Botulinum toxin-gold complexes have also been used to study the toxin internalization process into Torpedo synaptosomes . The images fit the three step sequence model in the pathway of botulinum neurotoxin poisoning.

J Neurol Sci, 1992 Jan, 107(1), 1 - 13
Disorders of neuromuscular transmission due to natural environmental toxins; Senanayake N et al.; A variety of natural toxins of animal, plant, and bacterial origin are capable of causing disorders of neuromuscular transmission . Animal toxins include venomous snakes and arthropods, venoms of certain marine creatures, skin secretions of dart-poison frogs, and poisonous fish, shellfish, and crabs . There are plant poisons such as curare, and bacterial poisons such as botulinum toxin . These act at single or multiple sites of the neuromuscular apparatus interfering with voltage-gated ion channels, acetylcholine release, depolarization of the postsynaptic membrane, or generation and spread of the muscle action potential . The specific actions of these toxins are being widely exploited in the study of neuromuscular physiology and pathology . Some toxins have proved to be valuable pharmaceutical agents . Poisoning by natural neurotoxins is an important public health hazard in many parts of the world, particularly in the tropics . Poisoning may occur by a bite or a sting of a venomous animal, or by the ingestion of poisonous fish, shellfish or other marine delicacies . Contaminated food is a vehicle for poisons such as botulinum toxin . Clinically, a cardinal feature in the symptomatology is muscle paralysis with a distribution characteristic of myasthenia gravis, affecting muscles innervated by cranial nerves, neck flexors, proximal limb muscles, and respiratory muscles . Respiratory paralysis may end fatally . This paper reviews from the clinical and pathophysiologic viewpoints, naturally occurring environmental neurotoxins acting at the neuromuscular junction.

Med Dosw Mikrobiol, 1992, 44(3-4), 201 - 5
{Evaluation of contents of A and B group substances in biological preparations}; Banach W; The study was aimed at determination of blood A and B group substances in biological preparations used in Poland . Twenty three series were investigated, namely: Di-Te-Per, Ty-Te, Ty, Te, against cholera, vaccine according to Delbet and Panodin . Also were tested: 65 series of imported preparations of immunoglobulin g (i.v.) such as Endobulin, Sandoglobulin, Gamma-Venin, Veinoglobuline and 5 local series such as Bioglobulin, as well as 9 series of preparation LNI (i.m.) Human Gamma Globulin . Presence of substance A was detected in tetanus and botulinum horse antitoxins in amount from 3.75 micrograms/ml to 30 micrograms/ml . Group substances A and B contained 6 series of LNI preparations-Veinoglobuline . Amount of substance A was detected as 3.75 micrograms/ml-7.5 micrograms/ml and of substance B as 2,5 micrograms/ml-5 micrograms/ml . Group substances A and B were not present in vaccines used according to vaccination calendar.

Brain Res, 1991 Dec 6, 566(1-2), 219 - 24
Muscle denervation increases thyrotropin-releasing hormone (TRH) biosynthesis in the rat medullary raphe; Van den Bergh P et al.; To determine whether thyrotropin-releasing hormone (TRH) could exert a trophic role in ventral horn motor neurons, we examined the effect of muscle denervation with botulinum toxin A on TRH mRNA in the rat medullary raphe by in situ hybridization histochemistry . Compared to controls, denervated rats showed a significant increase in the number and silver grain density of hybridized medullary raphe neurons . Increased proTRH gene expression in the medullary raphe in response to motor unit perturbation indicates that TRH may be trophic to lower motor neurons.

Arch Neurol, 1991 Dec, 48(12), 1253 - 6
Clinical correlates of response to botulinum toxin injections; Jankovic J et al.; We studied 242 patients with cervical dystonia who had adequate follow-up after botulinum toxin injections to determine which clinical variables had a predictive value in the treatment outcome . Twenty-one patients (16%) categorized as nonresponders were compared with 113 patients (47%) considered to be definite responders . On average, the nonresponders had symptoms for 14 years longer than responders . Seventy-eight of 100 patients with complications were female compared with 54% of 190 patients without complications . In addition, patients with complications weighed less than those without complications . Both findings suggest that the occurrence of complications is related to smaller mean neck muscle mass . Botulinum toxin antibodies were detected in 35.7% of the nonresponders tested and in none of the responders . This comprehensive analysis of outcome variables leads us to conclude that patients with a long duration of dystonia before their first botulinum toxin injection respond less well than those with a short duration of symptoms, that some patients lose their responsiveness because of the development of blocking antibodies, and that women are more likely to develop complications, such as dysphagia and neck weakness, than are men.

J Protein Chem, 1991 Dec, 10(6), 637 - 49
Botulinum neurotoxin type A: structure and interaction with the micellar concentration of SDS determined by FT-IR spectroscopy; Singh BR et al.; Secondary structures of botulinum neurotoxin type A have been determined using Fourier transform infrared spectroscopy in the amide I and amide III frequency regions . Using Fourier self-deconvolution, second derivatization, and curve-fit analysis, the amide I frequency contour was resolved into Gaussian bands at 1678, 1654, 1644, and 1634 cm-1 . In the amide III frequency region, several small bands were resolved between 1320 and 1225 cm-1 . Assignments of the bands in both amide I and amide III frequency regions to various types of secondary structures and the estimation of spectral band strengths by integrating areas under each band suggested that the neurotoxin contains 29% alpha-helix, 45-49% beta-sheets and 22-26% random coils . These values agreed very well with those determined earlier from CD spectra . The neurotoxin was treated with a micellar concentration of sodium dodecyl sulfate to simulate interaction between the protein and the amphipathic molecules . Sodium dodecyl sulfate micelles induced significant alterations both in the spectral band positions, and their strengths suggest refolding of the neurotoxin polypeptides . However, these changes were not entirely reversible, which could implicate the role of the altered structures in the function of the neurotoxin.

J Neurocytol, 1991 Dec, 20(12), 982 - 97
Formation of new muscle fibres and tumours after injection of cultured myogenic cells; Wernig A et al.; We examined the effects of implantation of cultured myogenic cells from a permanent cell line into soleus muscles of histocompatible adult mice . Myogenic cells (10(6) or 10(4)) were implanted into intact muscles, muscles frozen with liquid nitrogen, paralysed with botulinum toxin or reinnervated after long-term (seven months) denervation . Formation of numerous muscle fibres in myogenic cell-injected muscles raised the total number of fibres up to ten times above control by four weeks . Larger effects were found in freeze-damaged than in paralysed muscles . The new fibres had small calibers, considerable length (greater than 1.3 mm, maximum distance over which serial sections were made), were multinucleated and were oriented parallel to the large-diameter fibres of the host muscles . In some experiments beta-galactosidase, introduced into myogenic cells via retroviral transfection, was detected in small and large muscle fibres 4-20 weeks after implantation, indicating survival of the grafted cells and formation of mosaic (host-donor) and new fibres of donor origin . Muscle weight increased significantly and, rather surprisingly, a parallel increase was found in isometric tetanic tension of isolated nerve-muscle preparations; thus tension per mg muscle tissue was not different from normal . By eight weeks reduction of acetylcholine sensitivity and down-regulation of neural cell adhesion molecule to normal were observed, indicating that synaptic transmission at the new fibres was mature . After different periods of time (5-20 weeks, depending on the subclone used) tumours developed in most but not all injected limbs (37 out of 39) . The tumours were destructive to the muscles and were classified as rhabdomyosarcomas . Prior to tumour formation, neural cell adhesion molecule positive cells reappeared in the muscles; since the myogenic cells initially produced differentiated muscle fibres, it appears that malignant growth is induced by factors in vivo . Thus, at present the outcome of such implantation is unpredictable.

Br J Ophthalmol, 1991 Dec, 75(12), 737 - 9
Natural history of treatment of facial dyskinesias with botulinum toxin: a study of 50 consecutive patients over seven years; Mauriello JA et al.; To determine the long-term efficacy of botulinum toxin injections for the treatment of facial dyskinesias we studied 50 consecutive patients with blepharospasm, hemifacial spasm, and Meige syndrome . All received their first injection between September 1983 and June 1984 . A total of 520 injections were given; the average number of injections per patient was 10.4 over the seven-year period ending September 1990 . Twenty-six (52%) of the patients continued to return for periodic injections, while three patients no longer receive injections since they failed to respond adequately to treatment . Three patients with blepharospasm were in remission and required no further treatment, after a series of six, four, and three injections . Six patients were treated until they died of causes unrelated to facial dyskinesia or its treatment . Six patients are still being treated elsewhere because they could obtain injections closer to their homes . Five of the original 50 patients have been lost to follow-up . A patient with hemifacial spasm had one injection with good result but was not sufficiently bothered by her disease to return for reinjection . Complications were transient, minimal, well tolerated, and did not increase with increased number of injections.

Can J Ophthalmol, 1991 Dec, 26(7), 383 - 5
Temporary tarsorrhaphy induced with type A botulinum toxin; Wuebbolt GE et al.; We describe two children with corneal epithelial defects resistant to healing in whom protective temporary ptosis was induced with type A botulinum toxin injected to the levator palpebrae superioris . Epithelial healing was rapid, and no complications were noted . With further study this procedure may become useful in the treatment of corneal epithelial defects resistant to healing.

Neurology, 1991 Nov, 41(11), 1800 - 5
Botulinum-induced alteration of nerve-muscle interactions in the human orbicularis oculi following treatment for blepharospasm; Alderson K et al.; To assess longstanding alterations in human muscle innervation induced by botulinum toxin, we studied motor axons in the orbicularis oculi of nine patients previously injected with botulinum toxin for treatment of benign essential blepharospasm (BEB) . Compared with untreated BEB and normal orbicularis oculi, muscle exposed to botulinum toxin developed persistent and cumulative alterations of innervation, including (1) thin, unmyelinated axonal collaterals that contact muscle end plates, (2) an increased number of muscle fibers innervated by individual terminal motor axons, (3) a profusion of unmyelinated axonal sprouts that end blindly, (4) an increased range of end plate sizes, and (5) multiple end plates on individual muscle fibers . The findings suggest that axonal sprouts which develop after botulinum-toxin-induced functional denervation can form new end plates . A single muscle fiber may then be innervated at separate sites by more than one axon.

Laryngoscope, 1991 Nov, 101(11), 1216 - 8
Treatment of oromandibular dystonia with botulinum toxin; Hermanowicz N et al.; Botulinum toxin produces muscle weakness by inhibition of acetylcholine release at the neuromuscular junction . The toxin has been used successfully for symptomatic treatment of focal dystonias . Our experience in the use of botulinum toxin for the treatment of oromandibular dystonia in five patients is reported . Improvement following treatment was reported as marked by 1 patient, moderate by 1 patient, and mild by 3 patients . Similar improvement was noted by the examiners . One patient with mild cosmetic improvement developed significant dysphagia requiring feeding by a nasogastric tube for a 3-month period.

J R Soc Med, 1991 Nov, 84(11), 650 - 1
Low dose botulinum toxin in spasmodic torticollis; D'Costa DF et al.; Botulinum toxin has been successfully used to treat spasmodic torticollis . The optimum dosage is not clear and the recommended doses in the United Kingdom are 20-25 ng . We have used much lower doses (average 13 ng) without loss of efficacy and accompanied by a reduction in side effects . We treated 12 patients (eight women and four men) with a mean duration of torticollis of 4 years . Eleven of the 12 patients (91%) showed an improvement in total scores for pain and degree of head movement . The benefits appeared a week after treatment and lasted for 3 months . Side effects were minimal and transient . Our experience suggests low doses of the toxin may be equally efficacious.

Am J Ophthalmol, 1991 Oct 15, 112(4), 381 - 4
Treatment of unilateral acute sixth-nerve palsy with botulinum toxin; Metz HS et al.; We studied 29 consecutive patients with acute unilateral sixth-nerve palsy, who received botulinum toxin injection to the antagonist medial rectus muscle . The average interval between onset of palsy and treatment was 40 days and the mean follow-up from the last injection was 14 months . Before treatment, esotropia in the primary position ranged from 12 to 45 prism diopters and limitation to abduction in the affected eye ranged from -2 (approximately 15 degrees lateral to midline) to -6 (15 degrees nasal to midline) . After treatment, 22 of 29 patients (76%) had complete recovery of motility as determined by version testing . Of the seven patients with a residual abduction deficit, two had fusion in the primary position, three had fusion with prismatic correction, and two patients required subsequent surgery . Botulinum toxin injection seems to be an effective treatment option in cases of acute unilateral sixth-nerve palsy.

J Biol Chem, 1991 Oct 15, 266(29), 19276 - 82
Suppression of cytoskeletal rearrangement in activated human neutrophils by botulinum C2 toxin . Impact on cellular signal transduction; Grimminger F et al.; Botulinum C2 toxin, a binary toxin which selectively ADP-ribosylates nonmuscle G-actin, was used to evaluate the role of cytoskeletal rearrangement in ligand-evoked signal transduction and secretory processes in human neutrophils (polymorphonuclear leukocyte) . Preincubation with the combined toxin components reduced the basal F-actin content and nearly completely suppressed the actin assembly initiated by the peptide and lipid chemoattractants formyl-methionyl-leucyl-phenylalanine, platelet activating factor, and leukotriene B4 . Superoxide production and elastase secretion were increased markedly under these conditions . Concomitantly, ligand-elicited phosphoinositide hydrolysis was augmented with particular increase in inositol monophosphate . This was paralleled by a severalfold amplification of diacylglycerol formation and sustained elevation of cytosolic calcium . The toxin-effected amplification of postreceptor events and secretory responses was most pronounced in response to formyl-methionyl-leucyl-phenylalanine greater than platelet activating factor greater than leukotriene B4 . All metabolic and secretory effects in C2 toxin-pretreated cells were sensitive to pertussis toxin inhibition . In conjunction with the recent finding of unchanged formyl-methionyl-leucyl-phenylalanine receptor binding and dissociation dynamics under influence of C2 (Norgauer, J., Just, I., Aktories, K., and Sklar, L . A . (1989) J . Cell Biol . 109, 1133-1140), the present investigation suggests amplification of postreceptor events as a major mechanism underlying C2 toxin-related increase in polymorphonuclear leukocyte secretory responses . Cytoskeletal rearrangement, putatively linked to phosphoinositide turnover and calcium transients, thus appears to be operative in temporal and/or spatial limitation of chemoattractant-evoked cellular signal transduction.

Neurology, 1991 Oct, 41(10), 1677 - 9
Ear click in palatal tremor: its origin and treatment with botulinum toxin; Deuschl G et al.; We report the successful treatment of a rhythmic, continuing ear click in a patient with palatal tremor with local injections of botulinum toxin into the tensor veli palatini muscle . We could demonstrate that the ear click occurred during contraction of the tensor veli palatini, which opens the eustachian tube . Therefore, we believe that the clicking noise is due to the sudden breakdown of the surface tension within the eustachian tube . Our observations suggest that the ear click is due to rhythmic discharges of the trigeminal nucleus rather than the ambiguous nucleus.

J Neurochem, 1991 Oct, 57(4), 1413 - 21
Botulinum neurotoxin light chain inhibits norepinephrine secretion in PC12 cells at an intracellular membranous or cytoskeletal site; Lomneth R et al.; Botulinum neurotoxin (NT) is a potent inhibitor of neurotransmitter secretion, but its intracellular mechanism and site of action are unknown . In this study, the intracellular action of NT was investigated by rendering the secretory apparatus of PC12 cells accessible to macromolecules by a recently described "cell cracking" procedure . Soluble cytoplasmic factors were depleted from permeabilized cells by washing to generate cell "ghosts" which retained cellular structural components and intracellular organelles (including secretory granules) . The PC12 cell ghosts exhibited Ca(2+)-activated {3H}norepinephrine release which was enhanced by cytosolic proteins and MgATP . PC12 cell ghosts provide the opportunity to distinguish the intracellular action of NT on soluble cytoplasmic components versus structural cellular components . The 150-kDa NT and the 50-kDa light chain of serotypes E and B, and to a lesser extent type A, inhibited Ca(2+)-activated {3H}norepinephrine release in PC12 ghosts, but not in intact PC12 cells . The 100-kDa heavy chain had no effect . This indicates that NT acts at an intracellular site in these cells permeabilized by "cell cracking." The inhibition of secretion by NT was rapid and irreversible under the incubation conditions used . NT inhibition of {3H}-norepinephrine release from PC12 ghosts occurred in the absence of cytosolic proteins and MgATP and was not reversed by the addition of cytosolic proteins and MgATP, indicating that NT acts at an intracellular membranous or cytoskeletal site.

What Is Functional Genomics?, What Is Pcr?, What Is Fermentation?, What Is Protein?, What Is Dna?, e, Microbes, n, Microbe, a, Bacteria, s, Microorganisms, i, Microbiology, i, Staphylococcus aureus, c, Ciprofloxacin, c, Activated sludge, r, Antimicrobials, n, Yeasts, c, Bactericidal, n, Shigella, e, Escherichia coli, i, Listeriosis, a, Botulin, a, Microbial, o, Escherichia coli, i, Microflora, e, Agrobacterium, o, Antibiotics, s, Staphylococcus aureus, s, Brevibacteria, o, Bacteriological, a, Escherichia coli, e, Staphylococcus aureus, r, Culture medium

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005