J Neurosci, 1997 Oct 1, 17(19), 7190 - 202 Ca2+ or Sr2+ partially rescues synaptic transmission in hippocampal cultures treated with botulinum toxin A and C, but not tetanus toxin; Capogna M et al.; Botulinum (BoNT/A-G) and tetanus toxins (TeNT) are zinc endopeptidases that cleave proteins associated with presynaptic terminals (SNAP-25, syntaxin, or VAMP/synaptobrevin) and block neurotransmitter release . Treatment of hippocampal slice cultures with BoNT/A, BoNT/C, BoNT/E, or TeNT prevented the occurrence of spontaneous or miniature EPSCs (sEPSCs or mEPSCs) as well as the {Ca2+}o-independent increase in their frequency induced by phorbol ester, 0.5 nM alpha-latrotoxin, or sucrose . {Ca2+}o-independent and -dependent release thus requires that the target proteins of clostridial neurotoxins be uncleaved . In contrast, significant increases in mEPSC frequency were produced in BoNT-treated, but not TeNT-treated, cultures by application of the Ca2+ ionophore ionomycin in the presence of 10 mM {Ca2+}o . The frequency of sEPSCs was increased in BoNT-treated, but not TeNT-treated, cultures by increasing {Ca2+}o from 2.8 to 5-10 mM or by applying 5 mM Sr2+ . Large Ca2+ and Sr2+ influxes thus can rescue release after BoNT treatment, albeit less than in control cultures . The nature of the toxin-induced modification of Ca2+-dependent release was assessed by recordings from monosynaptically coupled CA3 cell pairs . The paired-pulse ratio of unitary EPSCs evoked by two presynaptic action potentials in close succession was 0.5 in control cultures, but it was 1.4 and 1.2 in BoNT/A- or BoNT/C-treated cultures when recorded in 10 mM {Ca2+}o . Log-log plots of unitary EPSC amplitude versus {Ca2+}o were shifted toward higher {Ca2+}o in BoNT/A- or BoNT/C-treated cultures, but their slope was unchanged and the maximal EPSC amplitudes were reduced . We conclude that BoNTs reduce the Ca2+ sensitivity of the exocytotic machinery and the number of quanta released. Exp Neurol, 1997 Sep, 147(1), 96 - 102 Botulinum A toxin therapy: neutralizing and nonneutralizing antibodies--therapeutic consequences; Goschel H et al.; Although muscle-relaxant doses of botulinum A toxin (BoNT/A) are generally lower than doses stimulating the immune system, specific antibodies are raised in a substantial number of patients . As a rule, this necessitates the termination of treatment . Therefore, a reliable determination of specific anti-BoNT/A antibodies is helpful and we introduced, for this purpose, a novel in vitro toxin-neutralizing assay based on a nerve-muscle preparation . We measured the antibody titers in four groups of subjects: Group 1 comprised 75 randomly selected patients of a total of 295 who responded to treatment with Dysport in our local clinic . Five patients, in group 2, were nonresponders . Group 3 consisted of 32 untreated volunteers and group 4 of 8 subjects immunized with a toxoid more than 10 years ago . Two of the responders had marginal titers of neutralizing antibodies, while they were present in all nonresponders . The sera of all responders were also tested for nonneutralizing antibodies by ELISA . Their occurrence, however, was of no consequence to the therapeutic success . The blood samples of volunteers were free from specific antibodies, whereas antibodies persisted in the immunized subjects for longer than a decade . Patients from various clinics who had been treated unsuccessfully with the toxin-14 patients had received BOTOX, 7 had been treated with Dysport, and 7 with both products-all had neutralizing antibodies . Whether there was an antibody response depended on the amount of toxin administered . We believe, however, the effective toxin dose can be reduced by so much as to make antibody production highly improbable. J Rheumatol, 1997 Sep, 24(9), 1842 - 3 Botulinum toxin increases tearing in patients with Sjögren's syndrome: a preliminary report; Spiera H et al.; Three patients with Sjogren's syndrome (SS) who had severe xerophthalmia and blepharospasm received botulinum toxin injections for the treatment of their blepharospasm . They had a remarkable increase in tearing, measured by Schirmer's test, and a decrease in signs and symptoms of dry eyes after botulinum toxin injection periorbitally for blepharospasm . The mechanism for this increased tearing is unclear, but suggests a potential treatment for patients with severe xerophthalmia with SS. Infect Immun, 1997 Sep, 65(9), 3743 - 52 Molecular characterization of murine humoral immune response to botulinum neurotoxin type A binding domain as assessed by using phage antibody libraries; Amersdorfer P et al.; To produce antibodies capable of neutralizing botulinum neurotoxin type A (BoNT/A), the murine humoral immune response to BoNT/A binding domain (H(C)) was characterized at the molecular level by using phage antibody libraries . Mice were immunized with BoNT/A H(C), the spleens were harvested, and single-chain Fv (scFv) phage antibody libraries were constructed from the immunoglobulin heavy and light chain variable region genes . Phage expressing BoNT/A binding scFv were isolated by selection on immobilized BoNT/A and BoNT/A H(C) . Twenty-eight unique BoNT/A H(C) binding scFv were identified by enzyme-linked immunosorbent assay and DNA sequencing . Epitope mapping using surface plasmon resonance in a BIAcore revealed that the 28 scFv bound to only 4 nonoverlapping epitopes with equilibrium constants (Kd) ranging from 7.3 x 10(-8) to 1.1 x 10(-9) M . In a mouse hemidiaphragm assay, scFv binding epitopes 1 and 2 significantly prolonged the time to neuroparalysis, 1.5- and 2.7-fold, respectively, compared to toxin control . scFv binding to epitopes 3 and 4 showed no protection against neuroparalysis . A combination of scFv binding epitopes 1 and 2 had an additive effect on time to neuroparalysis, which increased to 3.9-fold compared to the control . The results suggest that there are two "productive" receptor binding sites on H(C) which lead to toxin internalization and toxicity . Blockade of these two epitopes with monoclonal antibodies may provide effective immunoprophylaxis or therapy against BoNT/A intoxication. Biochem Biophys Res Commun, 1997 Aug 18, 237(2), 388 - 93 Botulinum E toxin light chain does not cleave SNAP-23 and only partially impairs insulin stimulation of GLUT4 translocation in 3T3-L1 cells; Macaulay SL et al.; The stimulation of glucose uptake into fat and muscle by insulin results predominantly from the translocation of the glucose transporter, GLUT4, from an intracellular vesicle pool to the cell surface . Homologues of several key proteins known to be involved in the process of synaptic vesicle fusion have been identified on GLUT4 vesicles, including VAMP2 and cellubrevin . Syntaxin 4, SNAP-23 and/or SNAP-25 are also implicated in this process . Bacterial toxins that specifically cleave these proteins have been utilised to assess their involvement in cell function . We aimed to distinguish which of the SNAP isoforms are specifically involved in GLUT4 translocation . Here we show that both human (h) and mouse (m) SNAP-23, unlike SNAP-25, are not substrates for Botulinum E toxin light chain (BoNT/E) . Furthermore, we demonstrate that microinjection of differentiated 3T3-L1 cells with BoNT/E inhibited insulin stimulation of GLUT4 translocation only slightly, 27%, whereas tetanus toxin light chain, that cleaves VAMP2, inhibited insulin stimulation of GLUT4 translocation by 80% . These studies therefore do not support a major role for SNAP-25 in insulin stimulation of GLUT4 translocation and place SNAP-23 as a prime candidate for a role in this process. JAMA, 1997 Aug 6, 278(5), 418 - 24 Iraq's biological weapons . The past as future? Zilinskas RA. Between 1985 and April 1991, Iraq developed anthrax, botulinum toxin, and aflatoxin for biological warfare; 200 bombs and 25 ballistic missiles laden with biological agents were deployed by the time Operation Desert Storm occurred . Although cause for concern, if used during the Persian Gulf War, Iraq's biological warfare arsenal probably would have been militarily ineffective for 3 reasons: (1) it was small; (2) payload dispersal mechanisms were inefficient; and (3) coalition forces dominated the theater of war (ie, they had overwhelming air superiority and had crippled Iraq's command and control capability) . Despite the Gulf War defeat, the Iraqi biological warfare threat has not been extinguished . Saddam Hussein remains in power, and his desire to acquire weapons of mass destruction continues unabated . In this context, the international community must be firm in its enforcement of United Nations resolutions designed to deter Iraq from reacquiring biological warfare capability and must take steps to develop a multidisciplinary approach to limiting future development of weapons of mass destruction. Graefes Arch Clin Exp Ophthalmol, 1997 Aug, 235(8), 486 - 9 Frontalis suspension in the treatment of essential blepharospasm unresponsive to botulinum-toxin therapy: long-term results; Roggenkamper P et al.; Thirty-one patients with essential blepharospasm or lid opening disorder of the levator-inhibiting type, unresponsive to treatment with botulinum toxin, underwent frontalis suspension . Twenty-eight patients received bilateral surgery (three patients with bilateral complaints of different severity were operated on the more affected side; these patients were not included in the statistical analysis) . The mean age was 62.4 years +/- 8.52 (range 42-80 years) . The individual improvement of complaints was assessed by the patients using a percentage scale (0% = no improvement; 100% = no complaints) . Objective and subjective improvement was achieved in 26 of 28 patients . The mean subjective improvement was 57.7% +/- 31.4 . In 23 cases an additional treatment with botulinum toxin was administered . During follow-up period (mean 22.1 months +/- 11.6; range 5-40 months) the effect of surgery remained stable . There were no serious complications, in a 5 of 56 operated eyes suture granuloma had developed . Unlike other surgeries for treatment of blepharospasm (excision of the orbicularis muscle, resection of facial nerve branches) frontalis suspension can be considered as a minimally invasive, but very effective and (if desired) reversible procedure . Moreover, additional treatment with botulinum toxin can bring about further improvement. Eur J Neurosci, 1997 Aug, 9(8), 1773 - 7 Distinct effects of clostridial toxins on activity-dependent modulation of autaptic responses in cultured hippocampal neurons; Owe-Larsson B et al.; Clostridial neurotoxins proteolyse specific proteins implicated in synaptic vesicle exocytosis, but their actions on the release machinery in functional synapses is not well understood . Here we examine the effects of botulinum toxin A (BoNT/A) and tetanus toxin (TeTx) on autaptic transmission in cultured rat hippocampal neurons using whole-cell voltage clamp recordings . The proportion of cells responding to stimulation with an excitatory postsynaptic current (EPSC) and the magnitude of the remaining responses decreased gradually with increasing concentration of either toxin . However, the activity-dependent modulation (5 Hz repetitive stimulation) of EPSCs remaining after toxin inhibition differed markedly between the two toxins . The TeTx inhibition was associated with a persistent activity-dependent depression similar to that in control cells . In contrast, the BoNT/A inhibition was accompanied by a reversal of the modulation into facilitation, resembling that induced by lowering of the calcium concentration . These results demonstrate a difference between BoNT/A and TeTx in their mode of inhibition of synaptic vesicle exocytosis, which suggests that they exert their preferential actions at distinct steps of the release process. Toxicon, 1997 Aug, 35(8), 1337 - 40 Interaction between botulinum neurotoxin type A and ganglioside: ganglioside inactivates the neurotoxin and quenches its tryptophan fluorescence; Kamata Y et al.; This study found that ganglioside quenched the tryptophan fluorescence of botulinum neurotoxin type A (BoNT A), accompanied by the inactivation of the toxin under low ionic strength conditions . This finding suggests that the ganglioside-binding site of BoNT A contains tryptophan residues . The quantum yield (a conformation parameter) in BoNT A under high ionic strength conditions differed from that under low ionic strength . This observation indicates that high ionic strength may alter the conformation of BoNT A, resulting in failure of the interaction between BoNT A and ganglioside. Am J Physiol, 1997 Aug, 273(2 Pt 2), F283 - 8 Role of Rho and myosin phosphorylation in actin stress fiber assembly in mesangial cells; Kreisberg JI et al.; Treatment of renal glomerular mesangial cells with adenosine 3',5'-cyclic monophosphate (cAMP)-elevating agents induces actin stress fiber disassembly, myosin light chain (MLC) dephosphorylation, loss of adhesion to the substratum and cell shape change {J . I . Kreisberg and M . A . Venkatachalam . Am . J . Physiol . 251 (Cell Physiol . 20): C505-C511, 1986} . Thrombin and vasopressin block the effects of cAMP . Because these agents are known to promote stress fiber formation via the small GTP-binding protein Rho, we investigated the effect of an activated variant of Rho on the response to cAMP elevation . Microinjecting V14-Rho completely blocked the effect of cAMP elevation on cell shape and the actin cytoskeleton, whereas inactivating Rho with botulinum C3 exoenzyme induced stress fiber disruption and cell retraction that was indistinguishable from that caused by elevations in intracellular levels of cAMP . Disruption of actin stress fibers by cAMP has previously been ascribed to MLC dephosphorylation; however, both C3 and cytochalasin D also caused dephosphorylation of MLC, whereas blocking MLC dephosphorylation failed to block the cAMP-induced loss of actin stress fibers . We conclude that Rho can modulate the effects of cAMP elevation and suggest that MLC dephosphorylation may be a consequence of actin stress fiber disassembly. Biochem J, 1997 Aug 1, 325 ( Pt 3), 637 - 43 Characterization of the intracellular signalling pathways that underlie growth-factor-stimulated glucose transport in Xenopus oocytes: evidence for ras- and rho-dependent pathways of phosphatidylinositol 3-kinase activation; Thomson FJ et al.; The stimulation of glucose transport is one of the early cellular responses to growth factors and is essential for cell proliferation, yet the molecular processes that underlie this response are poorly defined . The aim of this study was to characterize the role of the low-molecular-mass G-proteins, Ras and Rho, and their downstream targets, Raf protein kinase and phosphatidylinositol 3-kinase, in the regulation of glucose transport in Xenopus oocytes by two distinct growth-factor receptors: the insulin-like growth factor I (IGF-I) tyrosine kinase receptor and the heterotrimeric G-protein-coupled lysophosphatidic acid (LPA) receptor . Microinjection of a neutralizing anti-Ras antibody partially blocked IGF-I-stimulated deoxyglucose uptake but was without effect on LPA-stimulated deoxyglucose uptake . In contrast, microinjection of the C3 coenzyme of botulinum toxin, which selectively ADP-ribosylates and inactivates Rho, inhibited LPA-stimulated, but not IGF-I-stimulated, deoxyglucose uptake . Similarly, LPA- but not IGF-I-stimulated deoxyglucose uptake was attenuated in oocytes expressing a dominant negative rho construct . Cells expressing a dominant negative mutant of Raf protein kinase exhibited markedly reduced sensitivity to both LPA and IGF-I, consistent with a role for endogenous Raf in glucose uptake by both growth factors . Furthermore, expression of a constitutively activated form of raf-1 resulted in a growth-factor-independent increase in deoxyglucose uptake . Measurements of phosphatidylinositol 3-kinase activity in microinjected cells support the hypothesis that the IGF-I receptor stimulates glucose transport by a Ras-dependent activation of phosphatidylinositol 3-kinase, whereas the G-protein-coupled LPA receptor controls this response by a pathway that involves Rho-dependent activation of a distinct phosphatidylinositol 3-kinase . Thus we provide evidence for clear differences in the signalling pathways that control glucose transport by G-protein-coupled and tyrosine kinase growth-factor receptors . Furthermore this is the first demonstration that active Rho is involved in the signalling pathways that regulate glucose uptake in response to some growth factors. J Otolaryngol, 1997 Aug, 26(4), 273 - 6 Botulinum toxin for cricopharyngeal dysphagia: case reports of CT-guided injection; Atkinson SI et al.; OBJECTIVE: The National Institutes of Health have recognized the use of botulinum toxin (Botox) as a therapeutic agent to treat many ophthalmologic and otolaryngologic disorders . There are three reports in the literature regarding the use of Botox to treat cricopharyngeal dysphagia, all describing good results . In the larger study, the toxin was administered under general anaesthetic . This article discusses CT-guided injection of Botox: a relatively noninvasive, out-patient procedure . We also discuss failure of injection in one case and complications in another . CONCLUSION: The patients described experienced improvement in their dysphagia symptoms following injection. J Protein Chem, 1997 Aug, 16(6), 607 - 18 Predicting differential antigen-antibody contact regions based on solvent accessibility; Lebeda FJ et al.; A novel computational approach was examined for predicting epitopes from primary structures of the seven immunologically distinct botulinum neurotoxins (BoNT/A-G) and tetanus toxin (TeTX) . An artificial neural network {Rost and Sander (1994), Proteins 20, 216} was used to estimate residue solvent accessibilities in multiple aligned sequences . A similar network trained to predict secondary structures was also used to examine this protein family, whose tertiary fold is presently unknown . The algorithm was validated by showing that it was 80% accurate in determining the secondary structure of avian egg-white lysozyme and that it correctly identified highly solvent-exposed residues that correspond to the major contact regions of lysozyme-antibody cocrystals . When sequences of the heavy (H) chains of TeTX and BoNT/A-G were analyzed, this algorithm predicted that the most highly exposed regions were clustered at the sequentially nonconserved N- and C-termini {Lebeda and Olson (1994), Proteins 20, 293} . The secondary structures and the remaining highly solvent-accessible regions were, in contrast, predicted to be conserved . In experiments reported by others, H-chain fragments that induced immunological protection against BoNT/A overlap with these predicted most highly exposed regions . It is also known that the C-terminal halves of the TeTX and BoNT/A H-chains interfere with holotoxin binding to ectoacceptors on nerve endings . Thus, the present results provide a theoretical framework for predicting the sites that could assist in the development of genetically engineered vaccines and that could interact with neurally located toxin ectoacceptors . Finally, because the most highly solvent-exposed regions were not well conserved, it is hypothesized that nonconserved, potential contact sites partially account for the existence of different dominant binding regions for type-specific neutralizing antibodies. Muscle Nerve, 1997 Aug, 20(8), 1041 - 3 Flexor digitorum superficialis: locations of individual muscle bellies for botulinum toxin injections; Bickerton LE et al.; Twenty-two Flexor Digitorum Superficialis (FDS) muscles from 18 cadavers were dissected to find the "Optimal Injection Site" (OIS) for botulinum toxin injections to individual bellies of FDS . Coordinates are given as a percentage of the distance along a landmarking line from the medial epicondyle to the pisiform and in millimetres (mm) lateral to it . The OIS were: FDS2: 72%, 14 mm; FDS3: 54%, 17mm; FDS4: 49%, 7mm; FDS5: 76%, 6mm . OIS measurements guide the electromyographer to localize the targeted muscle belly. J Chromatogr B Biomed Sci Appl, 1997 Jul 18, 695(1), 67 - 75 Identification of a recombinant synaptobrevin-thioredoxin fusion protein by capillary zone electrophoresis using laser-induced fluorescence detection; Asermely KE et al.; Capillary zone electrophoresis (CZE) was utilized to identify a synaptobrevin-thioredoxin fusion protein (TSB-51) . TSB-51 is a substrate for cleavage by botulinum toxin B at the Q(76)-F(77) site . TSB-51 was derivatized with a fluorophore, CBQCA {3-(4-carboxy-benzoyl)-2-quinoline-carboxaldehyde}, for 4 h at room temperature . Optimal conditions for CZE separation of the TSB-51-CBQCA complex were determined: buffer (sodium borate), pH (9.0), applied voltage (25 kV), temperature (25 degrees C) and forward polarity . SDS-PAGE showed that TSB-51 had a molecular mass of approximately 19 kDa . The protein was transferred to PVDF membrane and sequenced by the Edman degradation method verifying the first twelve amino acids as SDKIIHLTDDSF . TSB-51 was also collected during CZE separation and subsequently sequenced yielding the first three amino acids as SDK . This CZE-LIF method coupled with the CBQCA derivatization, fraction collection and Edman sequencing allowed for identification of the recombinant protein, a fast separation run time and utilization of small volumes of peptide (1.5 ng protein/23.6 nl injection) . This method will be used for monitoring the endopeptidase activity of botulinum toxin B on TSB-51. Biochem Biophys Res Commun, 1997 Jul 9, 236(1), 184 - 8 Evidence that synaptobrevin is involved in fusion between synaptic vesicles and synaptic plasma membrane vesicles; Almeida MT et al.; We have developed a model system, consisting of rat brain synaptic vesicles and rat brain synaptic plasma membrane vesicles, to study the fusion process associated with the exocytotic release of neurotransmitters . Our results show a significant increase in the extent of fusion when the reaction takes place in cytosol compared to that obtained when fusion is carried out in buffer . This effect is mediated by cytosolic proteins, although N-ethylmaleimide-sensitive factor does not play a role in fusion . We also registered an almost complete inhibition of fusion when synaptic vesicles were pre-incubated with botulinum toxin B, indicating that synaptobrevin plays an important role in the coalescence of membrane lipids of the interacting membranes. Gut, 1997 Jul, 41(1), 87 - 92 Achalasia: outcome of patients treated with intrasphincteric injection of botulinum toxin; Cuilliere C et al.; BACKGROUND: To evaluate the safety and clinical efficacy of botulinum toxin (BT) in patients with achalasia followed up for six months . METHODS: Fifty five symptomatic patients with manometrically proven achalasia were included in a multicentre prospective trial . Before and two weeks and two months after intrasphincteric injection of BT, symptoms of dysphagia, regurgitation, and chest pain were scored on a 0-3 scale, and lower oesophageal sphincter pressure (LOSP) was assessed . The symptom score was determined again at six months, clinical improvement being defined by < or = 3, relapse by > 3, and failure as a relapse after two injections or loss to follow up . RESULTS: Except for transient chest or epigastric pain (22%), no side effects were observed . There was a significant decrease in LOSP after treatment . Symptom scores were significantly improved at two weeks (2.0 (SD 1.6)), two months (1.7 (1.8)), and six months (1.9 (2.0)) compared with pretreatment values (5.1 (1.8), p < 0.001) . At six months, 33 patients had clinical improvement (27 after one injection), 17 were considered failures, and five had just relapsed . Although there was a trend for age (older patients being more responsive), age, sex, prior duration of symptoms, initial symptom score, weight loss, LOSP, magnitude of oesophageal contractions, vigorous or non-vigorous achalasia, previous dilatations, and radiological features were not predictive of results . CONCLUSIONS: This multicentre series confirms that intrasphincteric injection of BT is a safe procedure, resulting in clinical improvement in 60% of patients with achalasia at six months . The therapeutic role of BT in achalasia needs further evaluation with regard to other alternatives. Hokkaido Igaku Zasshi, 1997 Jul, 72(4), 351 - 5 {Motor dysfunction in the aged--approach to involuntary movements}; Tashiro K; The clinical, pathophysiological and therapeutic approaches to the representative involuntary movements encountered in the aged are described . The prevalence rates of Parkinson disease and essential tremor are very high, and their diagnoses and treatments are quite important . Recent advances in treating Parkinson disease with anti-parkinsonian medications and essential tremor with beta-adrenergic blockers were presented . Blepharospasms, though uncommon, but occasionally seen in the aged persons, are disabled conditions . The botulinus toxin injections to the orbicularis oculi muscles proved to show dramatic therapeutic effects, greatly contributing to these patients' ADL . The importance of neuroscience in the coming 21st century is also stressed. J Pediatr Ophthalmol Strabismus, 1997 Jul-Aug, 34(4), 229 - 34 Diplopia following subcutaneous injections of botulinum A toxin for facial spasms; Wutthiphan S et al.; PURPOSE: To study the incidence, cause, recovery time, and prevention of diplopia following subcutaneous injection of botulinum A toxin for the treatment of facial spasms . METHODS: Patients who experienced diplopia after botulinum A toxin injections had their deviations examined in detail . When the muscle that caused diplopia was identifiable, the injection closest to that muscle was omitted in the next treatment in an attempt to prevent diplopia . RESULTS: Of 250 patients receiving about 1500 sets of injections, 25 (1.7%) incidents of diplopia occurred in 10 patients . Excluding two patients who declined further treatment after having diplopia on their first botulinum A toxin treatment, seven of the remaining eight patients had multiple incidents of diplopia . The most common pattern of diplopia was "uncertain diagnosis." The most common identifiable cause of diplopia was paresis of the inferior oblique muscle . Omission of the injection into the central portion of the lower eyelids in the next treatment prevented recurrence of diplopia in only one of the four patients . No significant correlation between botulinum A toxin doses injected and times to recovery was noted . CONCLUSIONS: Diplopia following botulinum A toxin treatment is uncommon . Seven patients (3% of patients studied) had 22 episodes of diplopia (88% of episodes) . When diplopia occurs, it tends to recur on reinjection, sometimes with a prolonged recovery time . This response may not be dose dependent . The extraocular muscles of some patients may be more susceptible to chemodenervation than others, or botulinum A toxin may diffuse to extraocular muscles more easily in some patients than in others. Toxicon, 1997 Jul, 35(7), 1089 - 100 Protection by the heavy metal chelator N,N,N',N'-tetrakis (2-pyridylmethyl)ethylenediamine (TPEN) against the lethal action of botulinum neurotoxin A and B; Adler M et al.; The ability of N,N,N',N'-tetrakis (2-pyridylmethyl)-ethyenediamine (TPEN) to protect against botulinum neurotoxin (BoNT) A and B was examined in vivo in mice . To determine the protective efficacy of TPEN, mice were injected i.p . with TPEN as a single bolus or as multiple injections 30 min before and 0, 2, 4 and 6 hr following i.v . challenges with BoNT-A or -B . TPEN treatment did not alter the 24 hr lethality of BoNT but did produce a significant delay in the time to death . For a moderate dose of serotype A (20 LD50), five divided doses of TPEN prolonged the time to death from 7.8 +/- 0.4 hr to 9.9 +/- 0.5 hr . For serotype B, examined under comparable conditions, the prolongation of the time to death was from 6.1 +/- 0.2 hr to 9.4 +/- 0.6 hr . The range of TPEN doses that could be examined in vivo was limited by its acute toxicity . Although low doses of TPEN (< or = 10 mg/kg) were well tolerated, higher doses (> or = 30 mg/kg) led to ataxia, loss of coordination, convulsions and death in 20.3 min or less . In clonal NG108-15 cells, TPEN was found to produce cytotoxicity as revealed by increases in the secretion of the marker enzyme lactate dehydrogenase (LDH), and enhanced reactivity with the vital dye trypan blue . From LDH concentration-response data determined 24 hr after addition of TPEN, the threshold concentration for observing cytotoxicity was 10 microM and the IC50 was 19.8 microM . At the highest TPEN concentration tested (100 microM), cytotoxicity was detected 8 hr after TPEN addition and increased in severity over a 3 day period . The cytotoxicity in NG108-15 cells appears to be distinct from the rapid-onset toxicity observed in whole animals . These results suggest that TPEN may be of potential benefit in delaying the lethal actions of BoNT-A and -B, but its use is limited by its initial and delayed toxicity . Since the therapeutic and toxic actions of TPEN are both related to zinc chelation, the use of TPEN would need to be restricted to low doses as part of a combination therapy. J Pediatr Surg, 1997 Jul, 32(7), 1059 - 61; discussion 1061-2 Preliminary experience with intrasphincteric botulinum toxin for persistent constipation after pull-through for Hirschsprung's disease; Langer JC et al.; Although most children who have Hirschsprung's disease have an excellent result after pull-through surgery, some experience persistent constipation caused by "internal sphincter achalasia." Anal myectomy has been advocated for this problem, but it results in permanent injury to the sphincter and is not universally effective . Botulinum toxin has been safely used to selectively and reversibly weaken a variety of voluntary muscles and sphincters in both adults and children . Injection of botulinum toxin into the internal anal sphincter (IAS) should theoretically produce the same functional result as anal myectomy without permanent sphincter injury . Four children aged 4 to 8 years presented with persistent constipation after a pull-through procedure for Hirschsprung's disease . Two had associated encopresis, both of whom had previous myectomies . The authors performed four-quadrant intrasphincteric botulinum toxin injection (total dose, 15 U) . Resting IAS pressure decreased in all children 4 to 8 weeks after injection . Patients have been followed up for 7 to 9 months . One child (with Down's syndrome) remained symptomatically unchanged . The other three families reported significant improvement in bowel function in their children . In two of these, there was a return of symptoms 6 months after injection; one child underwent reinjection with good results . Postinjection incontinence occurred in three children, but resolved after several weeks in the one who did not have encopresis before botulinum toxin injection . These preliminary results suggest that botulinum toxin may represent a less invasive alternative to anal myectomy for children who have severe constipation after surgery for Hirschsprung's disease . If myectomy is contemplated, botulinum toxin may also be useful as a means of predicting which children may benefit. Eur J Neurosci, 1997 Jul, 9(7), 1488 - 98 Peroxynitrite and nitric oxide donors induce neuronal apoptosis by eliciting autocrine excitotoxicity; Leist M et al.; Endogenous generation of nitric oxide and its congeners, including peroxynitrite (ONOO-), has been implicated in the mechanism of neuron loss in neurodegenerative diseases . Accordingly, nitric oxide donors and ONOO- can elicit both apoptosis and necrosis in neuron cultures . Here we show that nitric oxide donors and ONOO- are each able to trigger apoptosis of mouse cerebellar granule cells by an excitotoxic mechanism requiring exocytosis and NMDA receptor-mediated intracellular Ca2+ overload . This conclusion is supported by the following findings . Apoptosis was induced by various nitric oxide donors or by direct addition of ONOO- to differentiated cerebellar granule cell cultures that were sensitive to NMDA toxicity, but not in cerebellar granule cells that did not display NMDA-induced cell death (i.e . early days in culture) or in various glial cell populations . Donors of ONOO- or nitric oxide stimulated a sustained increase in intracellular Ca2+, which was prevented by inhibitors of NMDA receptors, such as MK-801 and 5-phospho-aminovaleric acid, or by dampening neuronal electrical activity with high concentrations of extracellular Mg2+ . Moreover, these treatments and the exposure of cerebellar granule cells in nominally Ca2+-free media prevented apoptotic cell death . Both the intracellular Ca2+ increase and apoptosis elicited by ONOO- or the nitric oxide donors were prevented by blocking exocytosis with tetanus toxin or botulinum neurotoxin C. Neurology, 1997 Jul, 49(1), 189 - 94 Human response to botulinum toxin injection: type B compared with type A; Sloop RR et al.; Despite the clinical potential of botulinum toxin type B (BTXB) for treating focal dystonia, hemifacial spasm, and other movement disorders, particularly in those resistant to botulinum toxin type A (BTXA), no objective human data exist to compare the muscle paralysis resulting from these two botulinum toxin subtypes . To objectively compare the human muscle paralysis resulting from intramuscular injections of BTXB with that from BTXA, we measured the extensor digitorum brevis (EDB) M wave amplitude four times before and six times after injection with 17 different doses of BTXB (from 1.25 to 480 units) in 17 healthy volunteers . This established a dose-response curve that we compared with the previously published BTXA dose-response curve . After the establishment of the dose-response curve, we injected 10 new volunteers with five different doses of BTXB and BTXA measuring EDB M wave amplitude 4 times before and 13 times over 57 weeks after injection . The volunteers were randomized by dose and received BTXA and BTXB in opposite EDB muscles . The effect of the toxin in all volunteers was expressed as percent decline in M wave amplitude postinjection (% paralysis) . The maximal paralysis 2 weeks postinjection with 320 to 480 mouse units (MU) of BTXB was 50 to 75%, whereas maximal paralysis was 70 to 80% with 7.5 to 10 MU of BTXA . Postexercise M wave facilitation on day 9 postinjection averaged 63% for BTXB and 20% for BTXA . Seven weeks postinjection, BTXB-induced paralysis had improved by 66% with complete improvement by 11 weeks postinjection, whereas BTXA-induced paralysis had improved by only 6% at 7 weeks, and at 57 weeks postinjection 22% of the original muscle paralysis was still present . Thus, human muscle paralysis resulting from BTXB injection is not as complete or long-lasting as that resulting from BTXA. Laryngoscope, 1997 Jul, 107(7), 948 - 53 Treatment of ventricular dysphonia with botulinum toxin; Kendall KA et al.; Ventricular dysphonia, traditionally known as dysphonia plica ventricularis, is a voicing disorder in which the false vocal folds are used as a vibratory source in addition to or instead of the true vocal folds . Traditional treatment of ventricular dysphonia has been voice therapy, which may be slow to produce results if the false fold activity masks an underlying problem of the true folds, is long standing, or has produced hypertrophy of the supraglottic structures . We present seven cases of ventricular dysphonia treated with botulinum toxin injection into the false vocal folds followed by speech therapy . The addition of botulinum toxin to the treatment regimen speeds recovery of normal voicing and allows immediate evaluation of dynamic true vocal fold function by the treating professional. J Neurochem, 1997 Jul, 69(1), 340 - 7 L-aspartate but not the D form is secreted through microvesicle-mediated exocytosis and is sequestered through Na+-dependent transporter in rat pinealocytes; Yatsushiro S et al.; Rat pinealocytes accumulate glutamate in microvesicles and secrete it through exocytosis so as to transmit signals intercellularly . Glutamate is involved in the negative regulation of norepinephrine-stimulated melatonin production . In this study, we found that aspartate is also released from cultured rat pinealocytes during the exocytosis of glutamate . The release of aspartate was triggered by addition of KCI or A23187 (a Ca2+ ionophore) in the presence of Ca2+ and was proportional to the amount of L-glutamate released . Furthermore, the release of aspartate was inhibited by both botulinum neurotoxin type E and L- or N-type voltage-gated Ca2+ channel blockers . Bay K 8644, an agonist for the L-type Ca2+ channel, stimulated the release of aspartate 2.1-fold . Immunohistochemical analyses with antibodies against aspartate and synaptophysin revealed that aspartate is colocalized with synaptophysin in a cultured pinealocyte . HPLC with fluorometric detection indicated that the released aspartate is of the L form, although pinealocytes also contain the D form in their cytoplasm, corresponding to approximately 30% of the total free aspartate . Radiolabeled L-aspartate was taken up by the microsomal fraction from bovine pineal glands in a Na+-dependent manner . The Na+-dependent uptake of L-aspartate was strongly inhibited by L-cysteine sulfinate, beta-hydroxyaspartate, and L-serine-O-sulfate, inhibitors for the Na+-dependent glutamate/aspartate transporter on the plasma membrane . Na+-dependent sequestration of L-aspartate was also observed in cultured rat pinealocytes, which was inhibited similarly by these transporter inhibitors . These results strongly suggest that L-aspartate is released through microvesicle-mediated exocytosis from pinealocytes and is taken up again through the Na+-dependent transporter at the plasma membrane . The possible role of L-aspartate as an intercellular chemical transmitter in the pineal gland is discussed. FEBS Lett, 1997 Jun 16, 409(3), 339 - 42 The interaction of synaptic vesicle-associated membrane protein/synaptobrevin with botulinum neurotoxins D and F; Pellizzari R et al.; Botulinum neurotoxins type D and F are zinc-endopeptidases with a unique specificity for VAMP/synaptobrevin, an essential component of the exocytosis apparatus . VAMP contains two copies of a nine residue motif, termed V1 and V2, which are determinants of the interaction with tetanus and botulinum B and G neurotoxins . Here, we show that V1 plays a major role in VAMP recognition by botulinum neurotoxins D and F and that V2 is also involved in F binding . Site-directed mutagenesis of V1 and V2 indicates that different residues are the determinants of the VAMP interaction with the two endopeptidases . The study of the VAMP-neurotoxins interaction suggest a pairing of the V1 and V2 segments. EMBO J, 1997 Jun 2, 16(11), 3044 - 56 p140mDia, a mammalian homolog of Drosophila diaphanous, is a target protein for Rho small GTPase and is a ligand for profilin; Watanabe N et al.; Rho small GTPase regulates cell morphology, adhesion and cytokinesis through the actin cytoskeleton . We have identified a protein, p140mDia, as a downstream effector of Rho . It is a mammalian homolog of Drosophila diaphanous, a protein required for cytokinesis, and belongs to a family of formin-related proteins containing repetitive polyproline stretches . p140mDia binds selectively to the GTP-bound form of Rho and also binds to profilin . p140mDia, profilin and RhoA are co-localized in the spreading lamellae of cultured fibroblasts . They are also co-localized in membrane ruffles of phorbol ester-stimulated sMDCK2 cells, which extend these structures in a Rho-dependent manner . The three proteins are recruited around phagocytic cups induced by fibronectin-coated beads . Their recruitment is not induced after Rho is inactivated by microinjection of botulinum C3 exoenzyme . Overexpression of p140mDia in COS-7 cells induced homogeneous actin filament formation . These results suggest that Rho regulates actin polymerization by targeting profilin via p140mDia beneath the specific plasma membranes. Fortschr Neurol Psychiatr, 1997 Jun, 65(6), 256 - 60 {A rare case of psychogenic dysarthrophonia or a complex dyskinesia of the pharynx?}; Kiese-Himmel C et al.; We present the symptomatology of a complex articulation-related dyskinesia of the pharynx with intermittent functional velopharyngeal insufficiency . It became permanent gradually after a writer's cramp had remitted without therapy . No final nosologic assignment can be made on the basis of the described observations in the individual case . The development of the disease in the 56-year old male patient ended in occupational disability . It suggests manifestation of psychopathological events in the patient's life history of threshold situations by premorbid psychic vulnerability . The possibility of the etiopathologic interaction of emotional and neurological factors cannot be excluded . However, therapy with local botulinum toxin injections proved to be just as unsuccessful as logopedic treatment. Immunol Invest, 1997 Jun, 26(4), 491 - 504 Localization of the regions on the C-terminal domain of the heavy chain of botulinum A recognized by T lymphocytes and by antibodies after immunization of mice with pentavalent toxoid; Rosenberg JS et al.; We have mapped the regions recognized by T and/or B cells (Abs) on the C-terminal domain (Hc) of the heavy chain of botulinum neurotoxin serotype A (BoNT/A) after immunization of two inbred mouse strains with pentavalent toxoid (BoNTs A, B, C, D and E) . Using a set of synthetic overlapping peptides, encompassing the entire Hc domain (residues 855-1296), we demonstrated that T cells of Balb/c (H-2d) mice, primed with one injection of toxoid, recognized two major regions within residues 897-915 and 939-957 . After multiple inoculations with toxoid, T cells of Balb/c expanded their recognition ability and responded very well to challenge with peptide 1261-1279 and moderately to stimulation with peptide 1149-1167 . Unlike Balb/c T cells, those of toxoid-primed SJL (H-2s) mice exhibited a more complex profile and responded to challenge with a large number of overlapping peptides . After one toxoid injection, however, three peptides, 897-915, 939-957/953-971 overlap and 1051-1069, were the most potent T cells stimulators . After three toxoid injections, peptides 897-915 and 1051-1069 remained immunodominant while the third region was shifted upstream to 925-943/939-957 overlap . The immunodominant epitope within peptide 897-915 was recognized exclusively by T cells, since no Abs were detected against this region . The Ab binding profiles of the two mouse strains were quite similar, showing only small quantitative differences . Both, Balb/c and SJL anti-toxoid Abs displayed strong binding mainly to peptide 1177-1195, followed by peptides 869-887/883-901 overlap and 1275-1296 . In addition, a significant amount of Balb/c anti-toxoid Abs was bound to peptide 1135-1153 . Unlike Balb/c Abs, that interacted weakly with peptides 995-1013 and 1051-1069, the anti-toxoid Abs of SJL mice exhibited strong binding toward both peptides . The results showed that, in a given strain, the regions recognized by anti-toxoid Abs and T cells may coincide or may be uniquely B or T cell determinants. Ann Surg, 1997 Jun, 225(6), 655 - 64; discussion 664-5 Laparoscopic Heller myotomy and fundoplication for achalasia; Hunter JG et al.; OBJECTIVE: The goal of this study was to review the authors' results with laparoscopic cardiomyotomy and partial fundoplication for achalasia . SUMMARY BACKGROUND DATA: Pneumatic dilatation and botulinum toxin (BOTOX) injection of the lower esophageal sphincter largely have replaced cardiomyotomy for treatment of achalasia . After a brief experience with a thoracoscopic approach, the authors elected to perform cardiomyotomy laparoscopically, in combination with a partial fundoplication (anterior or posterior) . PATIENTS AND METHODS: Forty patients were treated between July 1992 and November 1996 . Thirty patients had previous therapy of achalasia, 21 with pneumatic dilation, 1 with BOTOX, 6 with balloon and BOTOX, and 2 with transthoracic cardiomyotomy . Three patients had previous laparoscopic fundoplication for gastroesophageal reflux . Symptom scores (0 = none to 4 = disabling) were obtained before surgery and after surgery . Barium swallows and esophagogastroduodenoscopy were performed in all patients . Esophageal motility study was performed in 36 patients . Laparoscopic Heller myotomy and fundoplication was performed through five upper abdominal trocars . A 7-cm myotomy extended 6 cm above the GE junction and 1 cm below the GE junction . A posterior fundoplication was performed in 32 patients, anterior fundoplication in 7 patients, and no fundoplication in 1 patient . Statistical inference was performed with a Wilcoxon signed rank test . RESULTS: Mean operative duration was 199 +/- 36.2 minutes . Mean hospital stay was 2.75 days (range, 1-13 days) . Dysphagia was alleviated in all but four patients (90%), and regurgitation in all but two patients (95%) (p < 0.001) . Chest pain and heartburn improved significantly (p < 0.01) as well . Intraoperative complications included mucosal laceration in six patients and hypercarbia in one . Postoperative pneumonia developed in two patients, and one patient had moderate hemorrhage from an esophageal ulcer 2 weeks after surgery . CONCLUSIONS: Laparoscopic cardiomyotomy and fundoplication appears to provide definitive treatment of achalasia with rapid rehabilitation and few complications. J Leukoc Biol, 1997 Jun, 61(6), 703 - 11 Complex regulation of human neutrophil activation by actin filaments: dihydrocytochalasin B and botulinum C2 toxin uncover the existence of multiple cation entry pathways; Wenzel-Seifert K et al.; In human neutrophils, the chemotactic peptide, N-formyl-L-methionyl-L-leucyl-L-phenalalanine (fMLP), the Ca(2+)-ATPase inhibitor, thapsigargin, and the lectins, concanavalin A (Con A) and mistletoe lectin I (ML I), stimulate the entry of Ca2+ and Na+ with subsequent activation of exocytosis and superoxide anion (O2-) formation . We studied the role of actin in neutrophil activation . The actin filament-disrupting substances, dihydrocytochalasin B (dhCB) and botulinum C2 toxin (C2 toxin) potentiated fMLP- and lectin-stimulated Ca(2+)- and Na+ entry . Lectin-induced Mn2+ entry was enhanced by actin disruption, whereas fMLP-triggered Mn2+ entry was unaffected . dhCB and C2 toxin inhibited fMLP- and lectin-stimulated Ba2+ influx . The actin disrupters also inhibited fMLP- and ML I-induced Sr2+ influx, whereas Con A-stimulated Sr2+ entry was not influenced by dhCB and C2 toxin . Thapsigargin-stimulated cation entry was not altered by actin disruption . DhCB and botulinum C2 toxin potentiated lysozyme release induced by all four stimuli . Con A and ML I per se activated O2- formation only in the presence and not in the absence of dhCB . Con A potentiated the stimulatory effects of ML I on O2- formation in the presence of dhCB and primed neutrophils to respond to ML I in the absence of dhCB . Our data indicate the following: (1) dhCB and C2 toxin uncover the existence of multiple cation entry pathways in neutrophils; (2) actin disruption facilitates exocytosis and O2- formation by enhancement of Ca(2+)- and Na+ entry and by altering the function of proteins involved in activation of secretion and O2- formation; and (3) Con A and ML I, which possess different sugar specificities, activate different signaling pathways in neutrophils. J Pediatr Surg, 1997 Jun, 32(6), 916 - 7 Botulinum toxin use in pediatric esophageal achalasia: a case report; Walton JM et al.; Esophageal achalasia (EA) has been historically treated by esophageal dilatation or myotomy with or without fundoplication . Botulinum toxin (Botox-Allergan) use in pediatric EA has not been previously described . The authors' objective was to observe the efficacy of botulinum toxin injection into the lower esophageal sphincter (LES) for EA . An 11-year-old boy presented with a 9-month history of frequent pneumonia, productive cough, and a 1-year history of chest discomfort and odynophagia . Chest radiograph showed changes compatible with aspiration . Upper gastrointestinal (UGI) series showed typical narrowing of the LES, and 24-hour pH study showed no reflux . Esophageal manometry showed classic findings of achalasia . An upper gastrointestinal endoscopy was performed showing a huge volume of retained food . A direct four-quadrant injection was performed with a total of 100 U of botulinum toxin into the LES . UGI series showed improvement in esophageal emptying . Esophageal manometry showed impressive improvement in LES pressure (preinjection, 44.1 mm Hg to postinjection mean of 16.6 mm Hg), percent relaxation (preinjection, 30% to postinjection, 58.8%), and duration of relaxation (preinjection, 1.9 seconds to postinjection, 11 seconds) . The patient has not had any further respiratory symptoms, chest pain, or odynophagia in 8 months of follow-up . Botulinum toxin injection is simple and effective for EA and merits its study in a prospective manner in the pediatric population. Clin Neuropharmacol, 1997 Jun, 20(3), 195 - 203 Automatic EMG-guided botulinum toxin treatment of spasticity; Finsterer J et al.; Conventional electromyographic (EMG) guidance in botulinum toxin therapy can localize a muscle, but the amount of electrical activity is assessed only subjectively . We wanted to introduce a quantitative EMG criterion, according to which the decision for/against toxin application could be made . Turn/amplitude analysis (TAA) was applied to nine patients with severe paraspasticity (n = 5), right upper or lower limb spasticity (n = 3), or tetraspasticity (n = 1) before and after toxin administration . Muscles were selected for toxin application if both mean turns/second and mean amplitude/turn exceeded the level of 150 . A mean Dysport dose of 116 mouse units (mu) (range 40-240 mu) was administered to each of the 26 muscles that met the EMG criterion . Thirty days after the injection, activities of daily living, pain, and TAA count improved in 89%, tone in 78%, and range of motion in 56% of the patients by at least 1 point on corresponding 5-point rating scales . TAA provides a useful EMG criterion for/against botulinum toxin application . Muscle selection according to this criterion leads to a significant subjective and objective toxin effect . TAA is a valuable tool to determine the benefit of single and subsequent botulinum toxin injections in the treatment of spasticity. Laryngoscope, 1997 Jun, 107(6), 710 - 5 Contemporary management of the aging brow and forehead; Koch RJ et al.; Management of the aging brow and forehead has recently evolved based on available innovative technologies . Likewise, procedure-specific indications have changed based on collective surgical experiences . No longer is the approach based solely on hair pattern or degree of brow ptosis . Patients require varying combinations of brow elevation (prior to blepharoplasty), correction of brow asymmetries, and hairline-preserving forehead elevation . Some may only require excisional or paralytic procedures of the frontalis muscle (horizontal forehead creases), corrugator supercilii muscles (vertical glabellar furrows), and procerus muscle (horizontal glabellar furrows) . We present a 3-year experience using a problem-specific approach . This incorporates endoscopic technology, botulinum toxin type A purified neurotoxin complex (Botox, Allergan, Irvine, CA) intramuscular injection, and traditional procedures such as the coronal, pretrichial, midforehead, and direct browlift . Current indications, patient selection, and results are also discussed. Ophthal Plast Reconstr Surg, 1997 Jun, 13(2), 81 - 3 Preventing ptosis after botulinum treatment; Scott AB; In a series of 33 blepharospasm patients who had the side effect of ptosis following therapeutic botulinum toxin type A (Botox: Allergan, Inc., Irvine, CA, U.S.A.) injection, we administered 41 injections of human botulinum immune globulin (IG) following injections of the toxin to test the dosage and timing of IG injection and its effectiveness in limiting or avoiding ptosis . An IG dose of 3.2 x 10(-3) international units (IU) per unit of Botox was effective in blocking toxin effect when injected into the same tissue site within 4 hours . An IG dose of 1.6 x 10(-2) to 3.2 x 10(-2) into the levator of the eye having more frequent ptosis in 19 patients reduced the incidence of ptosis to 11% . The fellow (control) eye had a ptosis incidence of 37% . No orbital hemorrhage or other adverse effect occurred from the IG or its injection. J Voice, 1997 Jun, 11(2), 232 - 7 Adductor spasmodic dysphonia: case reports with acoustic analysis following botulinum toxin injection and acupuncture; Crevier-Buchman L et al.; We analyzed frequency and duration parameters of voice and speech in two men with adductor spasmodic dysphonia (SD) . One was treated with botulinum toxin injection; the other received acupuncture therapy . Improvement after acupuncture therapy in terms of standard deviation of fundamental frequency, acoustic perturbation measurements, durational measurements of voice and speech, and spectrographic analysis was comparable to the results achieved with botulinum toxin injection . Voice and speech parameters were stable 1 year after acupuncture therapy. Infect Immun, 1997 Jun, 65(6), 2225 - 32 Expression of botulinum toxin binding sites in Xenopus oocytes; Bakry NM et al.; The binding of iodinated botulinum toxin type B to nerve membranes was studied by using rat and mouse preparations . The toxin was examined both in the single-chain and in the proteolytically processed dichain form, and binding sites both in the spinal cord and in various brain regions were assayed . Rat and mouse brains possessed specific binding sites for botulinum toxin type B . The average Kd values for the various rat and mouse membrane preparations examined were 4.2 +/- 0.7 nM and 3.7 +/- 0.9 nM, respectively . The average Bmax values for the same tissue preparations were 7.3 +/- 0.7 pmol/mg of protein and 7.5 +/- 1.9 pmol/mg protein, respectively . The binding of botulinum toxin type B to rat brain membranes was not antagonized by a polyclonal antibody against the cytosolic domain of synaptotagmin 1 or by a monoclonal antibody directed against the luminal domain of synaptotagmin 1 . In addition, these antibodies did not protect the mouse phrenic nerve-hemidiaphragm from toxin-induced neuromuscular blockade . Extraction of whole-brain mRNA and injection into Xenopus oocytes led to expression of binding sites for botulinum toxin . Extraction and injection of cerebellar mRNA led to expression of a higher density of binding sites . The number of binding sites was not diminished when oocytes were pretreated with antibodies against the cytosolic and luminal domains of synaptotagmin 1 . These findings are likely to aid in the isolation, characterization, and reconstitution of toxin binding sites. Neuroscience, 1997 Jun, 78(3), 883 - 93 Sodium-dependent increase in quantal secretion induced by brevetoxin-3 in Ca2+-free medium is associated with depletion of synaptic vesicles and swelling of motor nerve terminals in situ; Meunier FA et al.; Brevetoxin-3 at nanomolar concentrations markedly enhanced spontaneous quantal transmitter release from neuromuscular junctions equilibrated in a Ca2+-free EGTA medium . After about 3 h, the sustained increase in miniature endplate potential frequency led to an exhaustion of transmitter release . This increase still occurred after loading the nerve terminals with the Ca2+ chelator bis-(aminophenoxy)ethanetetra-acetate or after pretreatment with various pharmacological agents known to prevent Ca2+ release from intracellular pools, but was completely prevented by the Na+ channel blocker tetrodotoxin . Brevetoxin-3 also increased miniature endplate potential frequency from junctions treated with botulinum type-A toxin, but to a smaller extent than at normal junctions . At normal junctions, brevetoxin-3 exposure for 2 h increased the three-dimensional projected area of living motor nerve terminals in situ by about 74% while at botulinum type-A poisoned junctions a similar toxin exposure caused only a 29% increase . Tetrodotoxin prevented such effects, indicating that they are related to both Na+ entry into the terminals and increased quantal transmitter release . Ultrastructural examination of nerve terminals from junctions exposed for 3 h to brevetoxin-3 revealed profound depletions of clear and large dense core synaptic vesicles and an increase in coated vesicles and axolemma infoldings . These results indicate that brevetoxin-3 impairs the recycling of clear synaptic vesicles and are consistent with our immunofluorescent observations showing that synaptophysin epitopes can be revealed without nerve terminal permeabilization . In contrast, no such changes were detected in nerve terminals poisoned with botulinum type-A toxin which, after 3 h exposure to brevetoxin-3, retained their synaptic vesicles and had a normal appearance . We conclude that tetrodotoxin-sensitive Na+ entry into motor nerve terminals induced by brevetoxin-3 triggers external Ca2+-independent asynchronous quantal transmitter release, blocks synaptic vesicle recycling and induces swelling of the terminals . We suggest that an excess of cytoplasmic Na+ per se can activate the asynchronous neurotransmitter release process. Tidsskr Nor Laegeforen, 1997 May 30, 117(14), 2022 - 4 {Treatment of spasticity with botulinum toxin}; Kerty E et al.; Spasticity is a velocity-dependent pathologic increase in muscle resistance to stretch, and occurs in a variety of neurologic disorders . We report our controlled open study using botulinum toxin A for treatment of adductor spasticity in five patients with advanced multiple sclerosis . Clinical evaluation of spasticity and stiffness of joints is based on the Ashworth Scale and grade of passive abduction . Three patients showed no response; the two others experienced an excellent and longlasting effect . We also describe briefly the different spastic conditions where this treatment has been used successfully. J Biol Chem, 1997 May 30, 272(22), 14447 - 53 Docked secretory vesicles undergo Ca2+-activated exocytosis in a cell-free system; Martin TF et al.; The Ca2+-activated fusion of secretory vesicles with the plasma membrane responsible for regulated neurotransmitter and hormone secretion has previously been studied in permeable neuroendocrine cells, where requirements for ATP and cytosolic proteins were identified . As reported here, Ca2+-activated fusion mechanisms are also preserved following cell homogenization . The release of norepinephrine (NE) and other vesicle constituents from a PC12 cell membrane fraction was activated by micromolar Ca2+ (EC50 approximately 3 microM) and exhibited a dependence upon MgATP and cytosol . Ca2+-dependent NE release was inhibited by botulinum neurotoxins and by CAPS (Ca2+-dependent activator protein for secretion) antibody implying that syntaxin, synaptobrevin, SNAP-25 (synaptosomal-associated protein of 25 kDa), and CAPS are required for regulated exocytosis in this system . The exocytosis-competent membrane fraction consisted of rapidly sedimenting dense core vesicles associated with plasma membrane fragments . Free vesicles did not release NE either in the absence or presence of plasma membranes, indicating that only docked vesicles were competent for exocytosis under the reconstitution conditions used . A cell-free system for Ca2+-activated fusion will facilitate studies on the roles of essential proteins such as syntaxin, synaptobrevin, SNAP-25, and CAPS that act at post-docking steps in the regulated exocytotic pathway. Tidsskr Nor Laegeforen, 1997 May 20, 117(13), 1889 - 91 {Treatment of spasmotic torticollis with botulinum toxin A . A 5-year experience}; Borgmann R; 60 patients have been treated with botulinumtoxin A for cervical dystonia since december 1990 . These patients have been seen at 472 visits and received at least two treatments . 55 (92%) patients noted improvement, and marked improvement was noted by 50 out of 60 patients (83%) . Further improvement after repeated treatments has been seen up to five years . Only one patient experienced no effect after several treatments with marked improvement . This might have been due to production of antibodies against the toxin . Side effects occurred after 9% of the toxin injections, and dysphagia was the symptom most often reported . The side effects were usually mild and transient, and never gave reason to terminate the treatment. Rev Prat, 1997 May 15, 47(10), 1088 - 93 {Dystonia}; Vidailhet M et al.; Dystonia can be considered either as a symptom, or as a disease . An initial classification of dystonia can be made according to the localization and the severity of the spasms or the associated movement disorders such as myoclonus . A second classification differentiates idiopathic dystonia and secondary dystonia . Personnel medical history, familial cases, neurological symptoms such as pyramidal, cerebellar, oculomotor signs are helpful clues in the diagnosis strategy . Drugs, botulinum toxin, physiotherapy are often combined symptomatic treatment regardless of the cause of dystonia. Biochem J, 1997 May 15, 324 ( Pt 1), 217 - 24 Functional studies in 3T3L1 cells support a role for SNARE proteins in insulin stimulation of GLUT4 translocation; Macaulay SL et al.; Insulin stimulation of glucose transport in the major insulin-responsive tissues results predominantly from the translocation to the cell surface of a particular glucose transporter isoform, GLUT4, residing normally under basal conditions in intracellular vesicular structures . Recent studies have identified the presence of vesicle-associated membrane protein (VAMP) 2, a protein involved in vesicular trafficking in secretory cell types, in the vesicles of insulin-sensitive cells that contain GLUT4 . The plasma membranes of insulin-responsive cells have also been shown to contain syntaxin 4 and the 25 kDa synaptosome-associated protein (SNAP-25), two proteins that form a complex with VAMP 2 . The potential functional involvement of VAMP 2, SNAP-25 and syntaxin 4 in the trafficking of GLUT4 was assessed in the present study by determining the effect on GLUT4 translocation of microinjection of toxins that specifically cleave VAMPs or SNAP-25, or microinjection of specific peptides from VAMP 2 and syntaxin 4 . Microinjection of tetanus toxin light chain or botulinum D toxin light chain resulted in an 80 and 61% inhibition respectively of insulin stimulation of GLUT4 translocation in 3T3L1 cells assessed using the plasma-membrane lawn assay . Botulinum A toxin light chain, which cleaves SNAP-25, was without effect . Microinjection of an N-terminal VAMP 2 peptide (residues 1-26) inhibited insulin stimulation of GLUT4 translocation by 54% . A syntaxin 4 peptide (residues 106-122) inhibited insulin stimulation of GLUT4 translocation by 40% whereas a syntaxin 1c peptide (residues 226-260) was without effect . These data taken together strongly suggest a role for VAMP 2 in GLUT4 trafficking and also for syntaxin 4 . They further indicate that the isoforms of SNAP-25 isolated to date that are sensitive to cleavage by botulinum A toxin light chain do not appear to be involved in GLUT4 translocation. Biochemistry, 1997 May 13, 36(19), 5719 - 28 Botulinum neurotoxin B inhibits insulin-stimulated glucose uptake into 3T3-L1 adipocytes and cleaves cellubrevin unlike type A toxin which failed to proteolyze the SNAP-23 present; Chen F et al.; Types A, B, and C1 botulinum neurotoxin (BoNT), a group of selective Zn2+-dependent endoproteases, have been instrumental in demonstrating that their respective substrates {synaptosomal-associated protein with Mr = 25 kDa (SNAP-25), synaptobrevin (Sbr), and syntaxin} are essential for regulated exocytosis from nerve terminals and neuroendocrine cells . The colocalization of Sbr, or its homologue cellubrevin (Cbr), in the majority of the glucose transporter-isotype 4 (GLUT4)-containing vesicles from adipocytes implicates their involvement in insulin-stimulated glucose uptake, which results in part from enhanced fusion of these vesicles with the plasmalemma . In this study, exposure of cultured 3T3-L1 adipocytes to BoNT/B in a low-ionic strength medium was found to block insulin-evoked glucose uptake by up to 64% . BoNT/B was shown by immunoblotting to cause extensive proteolysis of Cbr and Sbr resulting in a significant blockade of the insulin-stimulated translocation of GLUT4 to the plasmalemma . This establishes that these two toxin substrates contribute to the insulin-regulated fusion of GLUT4-containing vesicles with the plasmalemma, at least in this differentiated 3T3-L1 clone . Although SNAP-25 was not detectable in the differentiated adipocytes, its functional homologue SNAP-23 is abundant and largely confined to the plasmalemma . SNAP-23 proved to be resistant to cleavage by BoNT/A . Consistent with these results, type A did not block insulin-induced glucose uptake, precluding a demonstration of its likely importance in this process. Rev Neurol, 1997 May, 25(141), 728 - 39 {Cerebral palsy therapy}; Papazian O et al.; Unfortunately, in spite of the advances in foetal and perinatal medicine in the last twenty years, the incidence of cerebral palsy has remained unchanged (1.5-2.5 per 1000 live births) . It has even possibly risen slightly in premature babies of low birth weight, in parallel with the increased survival of these babies . In spite of modern techniques of rehabilitation, 25% of these patients cannot walk and 35% are mentally retarded . This costs society 5,000 million dollars annually, not counting the loss of opportunity and the emotional and economic burden imposed on these families . The development of new preventive measures such as the use of antagonists of the cortical excitatory amino acids (which when in excess may cause irreversible cerebral damage in cases of hypoxic-ischaemic encephalopathy of the new born) . Intramuscular botulinum toxin and continuous intrathecal baclofen seem to promise a reduction in the incidence and functional incapacity of cerebral palsy. Rev Esp Enferm Dig, 1997 May, 89(5), 367 - 74 Treatment of achalasia with botulinum toxin; Lopez P et al.; We studied the efficacy of local injections of botulinum toxin in the treatment of patients with achalasia . Four patients diagnosed of achalasia using manometric, radiologic and endoscopic criteria, were treated with botulinum toxin (80 U) injected directly into lower esophageal sphincter (LES), via a sclerotherapy injector . Response to treatment was assessed by changes in symptom scores and LES pressure . All determinations were repeated after 10, 30, 90, 120 and 180 days of treatment . The patients improved after the initial injection . This improvement was accompanied by improved relaxation of the LES . Two patients relapsed after 30 and 65 days and the other two patients remained symptom-free 5 months after treatment . CONCLUSION: Botulinum toxin is probably a safe and effective alternative for the treatment of achalasia and should be considered in patients in whom pneumatic dilation has failed or who are poor surgical candidates . Long-term evaluation of the safety and efficacy of botulinum toxin in the treatment of achalasia is required. Arch Phys Med Rehabil, 1997 May, 78(5), 525 - 9 Botulinum toxin treatment of apraxia of eyelid opening in progressive supranuclear palsy: report of two cases; Piccione F et al.; We report two patients, with postural instability and dystonic parkinsonism whose adjunctive disabling feature was blindness due to an inability to reopen the eyes after voluntary closure of the eyelids, as in apraxia of lid opening (ALO) . Supranuclear downgaze paresis permitted the diagnosis of progressive supranuclear palsy (PSP) in one case . Electromyographic studies showed a loss of normal reciprocal inhibition between the levator palpebrae and the pretarsal portion of the orbicularis oculi, with a cocontraction of these two antagonist muscles . The evoked blink reflex, tested with the paired shock technique, showed enhanced recovery of R2 response . Botulinum toxin A injections directed toward the junction of the preseptal and pretarsal parts of the palpebral orbicularis oculi muscle improved eyelid motility in both patients . Successive static and dynamic balance training and development of compensatory strategies for visual scanning deficits reduced gait imbalance, the number of falls, and the disability level as measured on the Northwestern University Disability Scale. South Med J, 1997 May, 90(5), 522 - 5 Tongue protrusion dystonia: treatment with botulinum toxin; Charles PD et al.; We report the treatment experience in a series of patients with involuntary tongue protrusion resulting from oromandibular dystonia (OMD) or Meige's syndrome . A retrospective analysis of clinical findings and results of treatment was conducted on patients treated at Vanderbilt University Medical Center between 1989 and 1995 . After unsuccessful treatment with conventional oral medications, nine patients having involuntary tongue protrusion resulting from OMD or Meige's syndrome were treated with botulinum toxin type A (BTX-A) injected into the genioglossus muscle at four sites via a submandibular approach . A marked reduction in tongue protrusion was achieved in six patients (67%) . Of 35 consecutive injections, 83% were successful at reducing tongue protrusion . Mild dysphagia complicated 14% of the injections . The average dose injected was 34 (+/- 3) units producing a 15 (+/- 2) week average duration of effect . Injection of the genioglossus with BTX-A may prove to be a valid treatment option for involuntary tongue protrusion related to OMD or Meige's syndrome . A double-blind, placebo-controlled trial is needed to better define efficacy and adverse events. Ophthalmology, 1997 May, 104(5), 865 - 8 Blepharospasm and hemifacial spasm . Randomized trial to determine the most appropriate location for botulinum toxin injections; Price J et al.; PURPOSE: The purpose of the study is to analyze the effectiveness and side effects of botulinum toxin using four different treatment site applications to determine the most successful treatment regime with the least side effects . METHODS: In a prospective trial, 92 patients (50 blepharospasm and 42 hemifacial spasm) were assigned randomly to 1 of 4 different treatment groups (standard {S}, brow {B}, inner orbital {IO}, or outer orbital {OO}) . Each treatment group had a different pattern of injection sites in the orbicularis . A total of 285 treatments were given, and the mean follow-up time was 16.4 months . RESULTS: In the blepharospasm group, patients assigned to the standard group had a significantly longer duration of effect than for those in the brow, inner orbital, and outer orbital groups (8.1 weeks compared with 4.5, 4.2, and 3.1 weeks, respectively; P < 0.001) . In the hemifacial spasm group, patients in the outer orbital group had significantly shorter duration of effect than those in standard, brow, or inner orbital group (7.2 weeks compared with 12.6, 12.8 and 10.4 weeks, respectively; P < 0.001) . The four major complications of botulinum toxin treatment were epiphora, ocular irritation, ptosis, and diplopia . The inner orbital treatment produced significantly more episodes of ptosis (13% of treatments) . However, the standard treatment produced the most epiphora and ocular irritation (18% of treatments) . CONCLUSIONS: The position of the injection sites around the orbicularis influences the effectiveness and side effects of botulinum toxin treatment for patients with blepharospasm and hemifacial spasm . The further the treatment is away from the eyelid margin, the lower the risk of ocular side effects . The standard treatment produces the longest duration of effect in the blepharospasm group but with the most transient ocular irritation and epiphora . In the hemifacial spasm group, the brow treatment has an equally long duration of effect as that of the standard treatment with fewer side effects. Neurology, 1997 May, 48(5), 1440 - 2 Muscle fiber atrophy in leg muscles after botulinum toxin type A treatment of cervical dystonia; Ansved T et al.; Previous electrophysiologic studies on the effects of local injections of botulinum toxin type A (BTX-A) have indicated impaired neuromuscular transmission in distant muscles . To further study possible distant effects of repeated BTX-A injections, we obtained percutaneous muscle biopsies of the vastus lateralis muscle from 11 patients with cervical dystonia . We examined the biopsies with histopathology and morphometry, and compared them with age-matched healthy controls . There was an increased frequency of angular atrophic type IIB fibers in the patient group, and the mean size of IIB fibers was significantly smaller (p < 0.05) . In addition, there was a negative correlation between accumulated dose of botulinum toxin and relative size of type IIA fibers (p < 0.05) . We postulate that the observed atrophy is due to distant effects of botulinum toxin causing progressive denervation-like changes in non-treated muscle . This observation calls for further, prospective studies of the long-term effects of the treatment. Ann Otol Rhinol Laryngol, 1997 May, 106(5), 422 - 31 Magnetic resonance imaging of the development of otitis media with effusion caused by functional obstruction of the eustachian tube; Alper CM et al.; In this study, magnetic resonance imaging (MRI) was used to define in vivo the effect of experimental functional obstruction of the eustachian tube (ET) on vascular permeability and the development of middle ear (ME) effusion . After collection of baseline data for ME pressure and MRI, the right tensor veli palatini muscle of 10 cynomolgus monkeys was injected with botulinum toxin A to induce ET obstruction . The left tensor veli palatini muscle was injected with saline in 4 monkeys . Right and left ME pressures and compliances were measured twice daily over a follow-up period of 36 days, and MRI scanning sessions including administration of a contrast agent, gadopentetate dimeglumine, were repeated on days 3, 6, 11, 15, 21, 29, and 36 in 6 animals and on days 15, 21, 29, and 36 in 4 animals . Two right ears did not develop underpressures, 5 developed persistent underpressures, and 3 developed underpressures that resolved . No changes in MRI parameters were noted for the ears that did not develop underpressures, but a progressive brightening of the ME on T2-weighted images, indicative of the development of inflammation and effusion, was noted for the others . Also, an increasing rate of transfer of the contrast agent between the vascular and ME compartments, indicative of increasing vascular permeability, was observed to track the temporal changes in ME pressure . These results support a causal relationship between ET dysfunction, ME underpressures, increased vascular permeability, and otitis media with effusion Neuroscience, 1997 May, 78(2), 469 - 79 Expression of neurotransmitter genes in rat spinal motoneurons after chemodenervation with botulinum toxin; Jung HH et al.; Botulinum toxin is widely used for the treatment of focal movement disorders, where chemodenervation is used to decrease hyperactivity in selected muscles . Beside a focal paresis, widespread effects on neuromuscular synaptic function have been demonstrated . However, reactions of motoneurons after neuromuscular chemodenervation without gross morphological lesions are largely unknown . Peripheral axotomy, in contrast, leads to profound changes in the expression of several genes, including those encoding neurotransmitters, in motoneurons . We therefore examined the expression of neurotransmitter genes in rat motoneurons six days after intramuscular botulinum toxin application in the right gastrocnemius muscle . Similar doses of botulinum toxin as used in human where injected . A focal bilateral increase in expression of the choline acetyltransferase gene and a widespread bilateral increase of the beta-calcitonin-gene-related peptide and the enkephalin genes was measured in motoneurons after botulinum toxin injection . Cholecystokinin had a lower expression after botulinum toxin injections . Growth-associated protein 43, nitric oxide synthase, somatostatin and proopiomelanocortin messenger RNA were not found in motoneurons of both groups . Our results demonstrate that changes in the expression of neurotransmitter genes in motoneurons also occur after chemodenervation but with different patterns to those found after mechanical nerve lesioning . These changes reflect focal and widespread modulative events . The knowledge of these events should lead to a better understanding of the focal paralysis and of the more widespread effects found in human after intramuscular injection of botulinum toxin. Muscle Nerve, 1997 May, 20(5), 593 - 8 Treatment of occupational cramp with botulinum toxin: diffusion of toxin to adjacent noninjected muscles; Ross MH et al.; Over a 5-year period, 40 patients, 11 with musician's and 29 with writer's cramp, were treated with botulinum toxin A using a precise injection technique in which the hollow-bore electromyography (EMG) needle was positioned by both standard EMG and by muscle twitch evoked by stimulating current passed through it . Moderate to complete improvement in dystonia occurred in 28 patients (70%) after the first injection and in 34 patients (85%) after the second injection with better outcome in nonmusicians than in musicians . Of note, weakness of uninjected muscles, immediately adjacent to those injected, was found in 25/40 patients (63%) . The most common patterns of toxin spread were from flexor digitorum sublimis to profundus, extensor carpi radialis to extensor digitorum communis, and extensor indicis proprius to extensor pollicis brevis . Spread to, and weakness of, adjacent uninjected muscles was a major factor contributing to suboptimal outcome in 6/39 (15%) such patients. Infect Immun, 1997 May, 65(5), 1626 - 30 Antibody mapping to domains of botulinum neurotoxin serotype A in the complexed and uncomplexed forms; Chen F et al.; The domain organization of the botulinum neurotoxin serotype A was studied by using antibody mapping of 44 monoclonal single-chain variable fragments . The analysis was carried out on (i) the individual domains of botulinum neurotoxin holotoxin (binding, translocation, and catalytic), (ii) botulinum neurotoxin holotoxin, (iii) the botulinum neurotoxin holotoxin in complex with the nontoxic portion, and (iv) botulinum neurotoxin holotoxin and nontoxic portion of the complex recombined in vitro . All 44 antibodies mapped to individual domains of botulinum neurotoxin . Forty of the 44 single-chain variable fragments bound the botulinum neurotoxin holotoxin relative to the isolated domains, suggesting that 4 epitopes are covered when the individual domains are in the holotoxin form . Only 20 of the antibodies showed a positive reaction to the toxin while in complex with the nontoxic portion . All of the covered epitopes were mapped to the binding domain of botulinum neurotoxin, which suggested that the binding domain is in direct contact with the nontoxic portion in the complex . Based on the antibody mapping to the different domains of the botulinum neurotoxin holotoxin and the entire complex, a model of the botulinum neurotoxin complex is proposed. J Neurosci, 1997 May 1, 17(9), 3254 - 61 Repeated cocaine modifies the mechanism by which amphetamine releases dopamine; Pierce RC et al.; This study determined whether daily cocaine administration initiates a calcium requirement for the increase in extracellular dopamine produced by psychostimulants . The increase in extracellular dopamine induced by perfusion of amphetamine through a microdialysis probe in the nucleus accumbens shell was enhanced in cocaine- relative to saline-pretreated rats . The augmented portion of the amphetamine-induced increase in nucleus accumbens dopamine was abolished by the coperfusion of L- or N-type calcium channel blockers . Inhibition of calcium/calmodulin-dependent protein kinase II (CaM-KII) also prevented the augmented increase in dopamine by amphetamine, whereas inhibition of vesicular exocytosis by botulinum toxin B was ineffective . When the concentration of extracellular dopamine in the nucleus accumbens was elevated by blocking the plasmallemal dopamine transporter with GBR-12909, the augmented increase in extracellular dopamine in rats sensitized to repeated cocaine was blocked by a CaM-KII inhibitor . Pretreatment with botulinum toxin B prevented the increase in extracellular dopamine by GBR-12909 in both cocaine-pretreated and control rats . Taken together, these results demonstrate that the psychostimulant-induced enhanced increase in extracellular dopamine in the nucleus accumbens shell of cocaine-pretreated rats arises from the induction of calcium- and CaM-KII-dependent mechanisms. J Neurosci, 1997 May 1, 17(9), 2967 - 79 Second messengers, trafficking-related proteins, and amino acid residues that contribute to the functional regulation of the rat brain GABA transporter GAT1; Quick MW et al.; Recent evidence indicates that several members of the Na+-coupled transporter family are regulated, and this regulation in part occurs by redistribution of transporters between intracellular locations and the plasma membrane . We elucidate components of this process for both wild-type and mutant GABA transporters (GAT1) expressed in Xenopus oocytes using a combination of uptake assays, immunoblots, and electrophysiological measurements of membrane capacitance, transport-associated currents, and GAT1-specific charge movements . At low GAT1 expression levels, activators of protein kinase C (PKC) induce redistribution of GAT1 from intracellular vesicles to the plasma membrane; at higher GAT1 expression levels, activators of PKC fail to induce this redistribution . However, coinjection of total rat brain mRNA with GAT1 permits PKC-mediated modulation at high transporter expression levels . This effect of brain mRNA on modulation is mimicked by coinjection of syntaxin 1a mRNA and is eliminated by injecting synaptophysin or syntaxin antisense oligonucleotides . Additionally, botulinum toxins, which inactivate proteins involved in vesicle release and recycling, reduce basal GAT1 expression and prevent PKC-induced translocation . Mutant GAT1 proteins, in which most or all of a leucine heptad repeat sequence was removed, display altered basal distribution and lack susceptibility to modulation by PKC, delineating one region of GAT1 necessary for its targeting . Thus, functional regulation of GAT1 in oocytes occurs via components common to transporters and to trafficking in both neural and non-neural cells, and suggests a relationship between factors that control neurotransmitter secretion and the components necessary for neurotransmitter uptake. Bol Asoc Med P R, 1997 Apr-Jun, 89(4-6), 57 - 9 Botulinum toxin A for the treatment of achalasia; Rodriguez Cruz E et al.; Pneumatic balloon dilatation of the esophagus is one of the current recommended treatment for achalasia . This procedure is associated with risks such as esophageal rupture . Surgery and percutaneous gastrostomy tube placement has been performed in severely affected individuals . The Botulinum Toxin A (BoTxA) is widely used to treat neuromuscular conditions in which spasticity is of concern . We present four cases in which BoTxA was used as an alternative of treatment and in which less invasive modalities were unsuccessful . The patients received a total of 80 units of BoTxA, applied to the submucosa in doses of 20 units in each predetermined quadrants to the lower esophageal sphincter . All patients demonstrated improvement of their symptoms without side effects in this study. Nervenarzt, 1997 Apr, 68(4), 346 - 50 {Neurogenic temporomandibular joint dislocation . Definition and therapy with botulinum toxin}; Daelen B et al.; Dislocation of the temporomandibular joint results from trauma, articulation disorders and changes in the equilibrium of the masticatory muscles . Previous classifications were mostly oriented on the interrelationships between the anatomical positions of the dislocated articular surfaces, like luxation, subluxation and discuss luxation . Etiologically, a distinction is made between traumatic and non-traumatic dislocation . Dislocation of the temporomandibular joint is described as a complication of a number of neurological diseases . We report on a patient who suffered recurrent dislocations of the temporomandibular joint as a complication of multiple sclerosis and whose luxations were successfully treated with botulinum toxin, thus warranting the introduction of the term "neurogenic dislocation of the temporomandibular joint". Rev Neurol, 1997 Apr, 25(140), 531 - 5 {Botulinum toxin in spastic infantile cerebral palsy: results in 27 cases during one year}; Sanchez-Carpintero R et al.; INTRODUCTION AND OBJECTIVES: Positive outcome of patients with spastic cerebral palsy treated with botulinum toxin reported in the last three years has led us to perform this study with the aim to show our experience in the management of spastic cerebral palsy with the toxin, determine its indications, analyze the results and propose new possible indications in the future . MATERIAL AND METHODS: We include 10 hemiplegic and 17 diplegic patients with an average age of 6 years and 7 months, followed up between 5 and 17 months . Clinical improvement was monitored using the PRS and EVFEL scales and articular motion range was measured 6 months before and after the injection while continuing physiotherapy . The injected muscles were adductor, hamstrings, triceps and posterior tibialis, and the doses were 1-2 U/muscle/kg body weight . RESULTS: The values on PRS improved an average of 24%, adductor angle 66% (p < 0.01), knee angle 40% (p = 0.05) and ankle angle 52% (p < 0.01); 96% of patients could get more physiological static or walking patterns because of the decrease of spasticity and those persisted after the effect of the toxin had worn off . It was maximum at 2 months, stabilized 4 to 6 months later and decreased during further 2 months . CONCLUSIONS: This experience leads us to propose higher starting dosage and to take into account the stability of postural pattern of each patient to choice the muscle to be injected . Other therapeutic possibilities are also proposed in children with fixed shortening e.g . combining the toxin with stretching casts. Med Hypotheses, 1997 Apr, 48(4), 337 - 9 Sialorrhea in amyotrophic lateral sclerosis: a hypothesis of a new treatment--botulinum toxin A injections of the parotid glands; Bushara KO; The inhibitory action of botulinum toxin is not confined to the neuromuscular junction . The toxin has long been known to block all the autonomic cholinergic fibers, including the major secretomotor parasympathetic fibers to salivary glands . The parotids are the largest of the salivary glands and their selective chemodenervation with botulinum toxin A is likely to result in substantial reduction of saliva production . Injection of the parotid glands with botulinum toxin is proposed as an new treatment for sialorrhea in patients with amyotrophic lateral sclerosis and other neurological diseases. Br J Dermatol, 1997 Apr, 136(4), 548 - 52 Double-blind trial of botulinum A toxin for the treatment of focal hyperhidrosis of the palms; Schnider P et al.; We performed a randomized double-blind study within-group comparison in 11 patients to study the effect of subcutaneous injections of botulinum A toxin in focal hyperhidrosis of the palms . A total dose of 120 mU (mouse units) of botulinum A toxin (Dysport) was injected into six different sites on one palm, whereas the other was injected with sterile saline . Objective quantification of sweat production was performed using digitized ninhydrin-stained sheets . Three weeks after treatment, the mean reduction of sweat production in the botulinum A toxin-treated palms was 26% (P < 0.001), after 8 weeks 26% (P = 0.002) and after 13 weeks 31% (P < 0.001) . Subjective assessment of sweat production by the patients using a visual analogue scale showed a 38% improvement in the botulinum A toxin-treated palms at 3 weeks (P = 0.002), 40% at 8 weeks (P = 0.002) and 38% at 13 weeks (P = 0.002) . Neither the objective measurement nor the subjective rating showed a statistically significant reduction of sweating in the placebo-treated palms . Three patients reported reversible minor weakness of powerful handgrip after injection at the toxin-treated site, lasting between 2 and 5 weeks. Brain, 1997 Apr, 120 ( Pt 4), 583 - 91 Postural responses to vibration of neck muscles in patients with idiopathic torticollis; Lekhel H et al.; Vibration of the dorsal muscles of the neck, simulating lengthening, in standing man causes a visible forwards tilt of the body shown on posturography as a tonic sagittal sway deviation . According to the theory that posture is organized with respect to a 'body schema' this deviation is a result of an interpretation of the concurrent neck afferent and vestibular signals . Considering the hypothesis that neck afferent signals may be misinterpreted in patients with spasmodic torticollis (ST) causing abnormal postural responses, we recorded body sway induced by unilateral dorsal neck muscle vibration in 22 idiopathic ST patients (19 treated with botulinum toxin) during upright stance with eyes closed . Comparison groups were 19 normal subjects and 11 patients with bilateral loss of vestibular function (labyrinthine defective, LD) in whom neck afference should be intact . Both treated and untreated ST and LD patients had absent or diminished sway deviations . When sway deviation did occur, it was sagitally oriented as with normal subjects and unrelated to ST head turns . In most ST and LD patients, neck vibration induced neck extension, an effect which is observed in normal subjects only if the torso is retrained . The results suggest that neck proprioceptive input retains local postural functions in ST, however, it is relatively ignored in the context of the whole body postural control and spatial orientation . The mild disorders of vestibular function reported in torticollis patients may be due to an inability to calibrate vestibular signals by reference to corroborative signals from neck proprioception. Brain, 1997 Apr, 120 ( Pt 4), 571 - 82 Botulinum toxin does not reverse the cortical dysfunction associated with writer's cramp . A PET study; Ceballos-Baumann AO et al.; Previous H2(15)O PET activation studies on patients with idiopathic torsion dystonia (ITD) have shown overactive striatum and frontal accessory areas and underactivity of the primary motor cortex and caudal supplementary motor area (SMA) during volitional movement . We have now examined activation of the motor system in healthy control subjects and patients with writer's cramp while they write a stereotyped word repetitively at a paced rate before and after treatment with botulinum toxin to see if these patients showed a similar pattern of abnormalities and whether they were reversible . As in ITD, our patients with writer's cramp showed impaired activation of the contralateral primary motor cortex, but enhanced activation of frontal association cortex . Botulinum-toxin treatment improved writing and increased activation in parietal cortex and caudal SMA . This may represent either a change in movement strategy or associated cortical reorganization secondary to deefferentation of alpha motor neurons . However, botulinum toxin failed to improve the impaired activation of the primary motor cortex . We conclude that, while botulinum toxin is clinically effective in writer's cramp, it does not reverse the associated dysfunction of primary motor and premotor cortex. Headache, 1997 Apr, 37(4), 253 - 5 Botulinum toxin injection for cervicogenic headache; Hobson DE et al.; We report a 28-year-old woman with a 5-year history of cervicogenic headache following a whiplash injury . Her unilateral neck pain, if aggravated by exertion, would create a predictable sequence of events leading to a hemicephalgia . She proved medically refractory to usual therapies, but had a striking response to a single botulinum toxin Injection in her symptomatic trapezius muscle . Repeated Injections every 3 months have been required to maintain this benefit . The Implications of this observation are discussed. Graefes Arch Clin Exp Ophthalmol, 1997 Apr, 235(4), 197 - 9 Comparison of two botulinum-toxin preparations in the treatment of essential blepharospasm; Nussgens Z et al.; A double-blind study was performed on 212 consecutive patients (58 men, 154 women) with essential blepharospasm, who received one injection of Botox and one injection of Dysport in two separate treatment sessions (at the first session the patients randomly received one of the drugs, at the second the other drug was given . The patients' mean age was 66.4 years +/- 8.14 (range 39-86 years) . The average dose of Botox per treatment was 45.4 IU +/- 13.3 (range 25-85 IU) and of Dysport 182.1 IU +/- 55.1 (range 100-340 IU) . We used an empirical ratio Botox:Dysport of 1:4 (IU) in order to ensure equal doses . All patients had received botulinum toxin injections prior to the present study (mean 15.3 injections +/- 9.4; range 1-43 injections) . The effect of Botox lasted 7.98 weeks +/- 3.8 (range 0-16 weeks), while the effect of Dysport lasted 8.03 weeks +/- 4.6 (range 0-22 weeks) . Side effects (ptosis, tearing, blurred vision, double vision, hematoma, foreign body sensation) were observed with Botox in 36 of 212 (17.0%) of the treatment sessions and with Dysport in 51 of 212 sessions (24.1%) . Ptosis was observed with Botox in 3 cases (1.4%) and with Dysport in 14 cases (6.6%) . There was no statistically significant difference in the duration of the treatment effect between the two preparations (P = 0.42) . The total number of side effects was lower with Botox than with Dysport; the significance of the difference was moderate (P < 0.05) . However, the rate of occurrence of ptosis was significantly lower with Botox (P < 0.01) . The bioequivalence, which varies between 1:3 and 1:6 (Botox:Dysport) in the literature, was found to be 1:4 in this study. Clin Plast Surg, 1997 Apr, 24(2), 199 - 212 Rejuvenation of the upper face . A logical gamut of surgical options; Michelow BJ et al.; The prime objective of forehead rejuvenation is to correct the factors that accentuate the impression of aging, namely, eyebrow ptosis, glabella frown lines, forehead wrinkles, and asymmetry . Options for forehead rejuvenation are undergoing a renaissance . With the wide variety of choices currently available, accurate analysis of the presenting problem is mandatory so that the appropriate surgical technique can be chosen . The less invasive endoscopic procedures have a low morbidity rate and great patient satisfaction . Not all patients are candidates for these procedures, however . A thorough knowledge of alternative methods, including botulinum injection, fat injection, fat grafting, and subcutaneous approaches to the brow, are necessary if optimal aesthetic results are to be accomplished . Having an armamentarium of forehead rejuvenation techniques is, therefore, the key to individualizing treatment for each patient. J Endocrinol, 1997 Apr, 153(1), R5 - 10 Involvement of SNAP-25 in TRH-induced exocytosis in pituitary GH4C1 cells; Masumoto N et al.; The synaptic membrane protein synaptosomal-associated protein (SNAP-25) has recently been implicated as one of the key proteins involved in exocytotic membrane fusion in neurons . However, the role of SNAP-25 in pituitary hormone release is not known . In this study, we determined that SNAP-25 is involved in regulated exocytosis in the clonal pituitary cell line GH4C1 . SNAP-25 messenger RNA and protein were detected in GH4C1 cells by RT-PCR and immunoblot analysis, respectively . Immunofluorescence analysis indicated that SNAP-25 protein was localized in the plasma membrane . Next, to determine the function of SNAP-25 in GH4C1 cells, specific inhibitors of SNAP-25, botulinum neurotoxin (BoNT)/A or /E, and antisense SNAP-25 oligonucleotide were used . Neither BoNT/A nor BoNT/E affected thyrotropin-releasing hormone (TRH)-induced cytosolic Ca2+ increase, but both inhibited TRH-induced exocytosis . Moreover, they dose-dependently inhibited TRH-induced prolactin release . The introduction of antisense oligonucleotide into the cells also inhibited TRH-induced prolactin release . These results suggest that SNAP-25 is involved in regulated exocytosis in GH4C1 cells. Arch Otolaryngol Head Neck Surg, 1997 Apr, 123(4), 389 - 92 The management of hyperfunctional facial lines with botulinum toxin . A collaborative study of 210 injection sites in 162 patients; Blitzer A et al.; OBJECTIVE: To determine the optimum dose and efficacy of botulinum toxin injections in the management of hyperfunctional facial lines . DESIGN: This study included 210 hyperfunctional facial sites in 162 different patients . The patients had preinjection and postinjection photographic documentation and ratings on a 4-point qualitative evaluation scale of lines at rest and with action . The patients then had botulinum toxin type A injections via a monopolar hollow bore, Teflon-coated electromyographic needle into the facial muscles associated with the hyperfunctional lines . The total dose for each region of 1.25 to 25 U was divided into 1.25- to 5-U aliquots representing 0.1 to 0.2 mL per injection site, depending on the site and the prior experience with that patient on using toxin . The patients had their reevaluation at 2 to 3 weeks after injection . Patients returned for further follow-up when the therapeutic effect diminished . PATIENTS: One hundred sixty-two patients had 210 hyperfunctional sites evaluated and injected . The group consisted of 25 male patients and 137 female patients ranging in age from 21 to 78 years with a mean (+/-SD) of 46.1 (+/-1.98) years . All patients had cosmetically troubling hyperfunctional lines involving the forehead, glabella, crow's feet (lateral canthal lines), nasolabial area, platysma, and mentalis region . RESULTS: All patients had an effect of toxin within the first 24 to 72 hours . Ninety-five percent of the patients treated had cosmetic improvement of unsightly facial lines or contractions . The best results were achieved in management of the forehead lines, followed by glabella, crow's feet, and nasolabial . The dose for forehead lines was 5 to 25 U (mean +/- SD, 17.3 +/- 6.2 U); glabellar lines, 5 to 20 U (mean +/- SD, 11.1 +/- 3.1 U); crow's feet, 5 to 15 U (mean +/- SD, 6.2 +/- 1.6 U); nasolabial, 2.5 to 5 U (mean +/- SD, 3.12 +/- 1.2 U); and platysma, 10 to 20 (mean +/- SD, 15 +/- 4.0 U) . Evaluation by age and site suggested a trend of increased toxin dose with increased age . Effects of the toxin are usually seen 24 to 72 hours after injection, and last from 3 to 6 months, whereon the increased muscular activity returns, as do the hyperfunctional lines . The only morbidity was related to temporary mild weakness of other adjacent facial muscles . There were no systemic side effects noted . CONCLUSION: Botulinum toxin is a safe and important adjunctive technique for the management of patients with symptomatic hyperfunctional facial lines. Drugs Aging, 1997 Apr, 10(4), 249 - 58 Gastrointestinal motility problems in patients with Parkinson's disease . Effects of antiparkinsonian treatment and guidelines for management; Jost WH; Gastrointestinal (GI) motility disorders are frequent in patients with Parkinson's disease, manifesting mainly as dysphagia, disorders of gastric emptying and constipation . The most likely causes of these disorders are cerebral degeneration and degeneration of the myenteric plexus . Although the effect of antiparkinsonian medication is largely overestimated, it certainly has an influence and should be adapted accordingly in patients with GI motility disorders . In particular, anticholinergic drugs should be avoided, and anamnesis, clinical examination and, if necessary, diagnostic tests performed . Domperidone, a peripheral dopamine antagonist, is the drug of choice for motility disorders of the upper GI tract, although cisapride is an alternative . In the lower GI tract, conservative therapeutic options should be used in the first instance . The administration of cisapride leads to a marked temporary improvement in symptoms in lower GI disorders, while rare forms of anism (involuntary dystonic contraction of the anal sphincter) may be treated with botulinum toxin. J Otolaryngol, 1997 Apr, 26(2), 92 - 6 Cosmetic upper-facial rejuvenation with botulinum; Ellis DA et al.; OBJECTIVE: This study was conducted to evaluate the cosmetic use of botulinum toxin type A (Botox), which blocks the release of acetylcholine at the presynaptic neuromuscular junction leading to an irreversible, but temporary chemical denervation muscular paralysis and weakness . This produces a significant cosmetic improvement of wrinkling in the upper face due to hyperfunctional animation . METHOD: A prospective clinical study representing our experience with this new technique is presented . Patient selection and evaluation, classification of animation lines, techniques, results and complications are discussed . In a 15-month period, 23 patients with seven anatomic sites were injected . Twenty-three patients had the lateral aspect and the inferior aspect of their squint lines injected, and 26 patients had their glabellar frownlines injected . RESULTS: Significant improvement occurred to the average depth and length of the glabellar frownlines . The subjective improvement by the patients was also significant . Regarding the crow's feet, the lateral canthal lines showed more improvement than the inferior lateral canthal lines because the latter has a greater component of zygomaticus major and minor muscle, which contributes to the inferior lateral squint line . CONCLUSION: Botox is a safe, easy-to-use, effective modality for the temporary elimination of hyperfunctioning upper-facial muscles. J Neurosurg Anesthesiol, 1997 Apr, 9(2), 149 - 53 Sensitivity to vecuronium after botulinum toxin administration; Fiacchino F et al.; When used to treat focal dystonias, botulinum toxin may cause a transient impairment of neuromuscular transmission in muscles distant from those injected . These systemic effects are not clinically evident, but should not be ignored when patients are exposed to other drugs or conditions that also impair neuromuscular transmission . A patient is described who underwent general anesthesia twice during treatment with botulinum toxin for a long history of blepharospasm . On both occasions, the neuromuscular block produced by vecuronium (0.05 mg kg-1) was monitored in the abductor digiti minimi muscle . Compared with that observed in 24 individuals who were free from neuromuscular problems, the patient's sensitivity to vecuronium was low 90 days after the seventh treatment with toxin and normal 8 days after the ninth . The possibility is considered that repeated treatments with the toxin may cause continuous remodeling of neuromuscular junctions and may cause the patient to develop some tolerance to the action of neuromuscular blockers. Plast Reconstr Surg, 1997 Apr, 99(4), 1006 - 11 Combination of hypoglossal-facial nerve anastomosis and botulinum-toxin injections to optimize mimic rehabilitation after removal of acoustic neurinomas; Laskawi R; This report refers to 10 patients treated with hypoglossal-facial nerve anastomosis and botulinum-toxin type A after removal of acoustic neurinomas . All patients who underwent this regimen showed reinnervation of mimic muscles after nerve anastomosis in all parts of the mimic musculature . Synkinesis could be observed in all patients . Botulinum-toxin injections into the orbicularis oculi muscle improved the functional outcome, leading to an approximation of the palpebral fissure width to the healthy side during heterogenous face movements . The mean duration of the toxin effect was 13.8 weeks . Side effects were rare . It can be stated that the combination of hypoglossal-facial nerve anastomosis and botulinum-toxin injections is suited to optimize mimic rehabilitation after removal of acoustic neurinomas. Biochemistry, 1997 Mar 18, 36(11), 3061 - 7 Importance of two adjacent C-terminal sequences of SNAP-25 in exocytosis from intact and permeabilized chromaffin cells revealed by inhibition with botulinum neurotoxins A and E; Lawrence GW et al.; Types A and E botulinum neurotoxin (BoNT) are Zn2+-requiring endoproteases which cleave nine and twenty-six residues, respectively, from the C-terminus of synaptosomal-associated protein of Mr = 25 kDa (SNAP-25) . Involvement of SNAP-25 in the exocytosis of large dense-core vesicles in bovine adrenochromaffin cells was examined by measuring cleavage of SNAP-25 in relation to the levels of Ca2+-evoked catecholamine release from cells exposed to BoNT/A or /E, either before or after permeabilization . The dose-dependency of inhibition of exocytosis correlated closely with the extents of SNAP-25 cleavage in cells permeabilized and then treated with BoNT/E . In intact cells exposed to 66 nM BoNT/A, virtually all of the SNAP-25 was truncated, accompanied by a near-complete inhibition of exocytosis; however, after their permeabilization a significant level of secretion was recorded upon Ca2+-stimulation . Importantly, this BoNT/A-resistant release from the permeabilized cells was dramatically lowered by subsequently adding BoNT/E, which further truncated the SNAP-25 fragment (lacking the C-terminal nine residues) that had been produced earlier by BoNT/A . Moreover, anti-SNAP-25 IgG decreased the BoNT/A-insensitive exocytosis . When permeabilized cells were exposed to either neurotoxin, both blocked MgATP-dependent secretion but only BoNT/E attenuated the energy-independent phase . These distinct inhibitory effects of the two neurotoxins demonstrate that residues 197-205 at the C-terminus of SNAP-25 are absolutely essential for exocytosis from intact cells whereas even after their removal a significant proportion of the exocytotic response can be elicited from permeabilized cells, but this is reliant on amino acids 180-196 . Moreover, the latter but not residues 197-205 are implicated in a late, MgATP-independent step of exocytosis, which is blocked by BoNT/E but nonsusceptible to BoNT/A. J Neurosci, 1997 Mar 15, 17(6), 1898 - 910 Inhibition of transmitter release correlates with the proteolytic activity of tetanus toxin and botulinus toxin A in individual cultured synapses of Hirudo medicinalis; Bruns D et al.; We have studied the effects of tetanus toxin and botulinus toxin A on neurotransmitter release in the Retzius-->P-cell synapse of the leech and exploited the unique properties of this system, which allow for combined physiological and biochemical analyses in single-cell pairs . The sequences of Hirudo medicinalis synaptobrevin and synaptosomal-associated protein of 25 kDa (SNAP-25), deduced by cDNA cloning, are 61 and 55% identical, respectively, to their corresponding mammalian homologs . Whereas Hirudo synaptobrevin is proteolyzed by tetanus toxin, its SNAP-25 isoform is resistant to botulinus toxin A cleavage because of amino acid substitutions within and around the putative cleavage site . In close correlation, microinjection of tetanus toxin into the presynaptic neuron produced a block of transmitter release, whereas botulinus toxin A had no effect on synaptic transmission . Subsequent immunoblotting of single-cell pairs demonstrated directly that the tetanus toxin-mediated block of exocytosis is accompanied by cleavage of synaptobrevin in the injected neuron, resulting in the generation of a detectable C-terminal cleavage product . Immunoblotting also confirmed the resistance of SNAP-25 to botulinus toxin A cleavage in vivo . Using recombinant proteins, we show that the N-terminal fragment of synaptobrevin released by tetanus toxin, but not its C-terminal membrane-anchored cleavage product, participates with syntaxin and SNAP-25 in synaptic SNAP receptor (SNARE) ternary complex formation in Hirudo . Our data demonstrate a direct correlation between the inhibition of transmitter release and the ability of the neurotoxin to proteolyze its target protein and support the view that SNARE ternary complex formation is an important step leading to synaptic vesicle exocytosis. Neurosci Lett, 1997 Mar 14, 224(2), 91 - 4 Botulinum neurotoxin serotype C: a novel effective botulinum toxin therapy in human; Eleopra R et al.; Botulinum neurotoxin (BoNT) serotype A is commonly used in the treatment of focal dystonia . Nevertheless, some patients are or become resistant to this serotype . Consequently, other different serotypes have to be used . A comparison of the neuromuscular blockade induced by BoNT type A and C in the extensor digitorum brevis muscles of voluntary subjects was studied, by evaluating the amplitude variation over the time (until 90 days) of the compound muscular action potential elicited by supramaximal electrical stimulation of the peroneal nerve at the ankle . A very similar effect and temporal profile, was observed for each serotype . On this basis, two patients with idiopathic facial hemispasm and one with blepharospasm were treated with BoNT serotype C with very beneficial long lasting effects. Brain Res, 1997 Mar 7, 750(1-2), 41 - 7 Cultured astrocytes express proteins involved in vesicular glutamate release; Jeftinija SD et al.; Bradykinin induces receptor-mediated calcium-dependent release of glutamate from cultured astrocytes through a mechanism that is neither due to cell-swelling mechanism nor due to the reversal of the glutamate transporter . Astrocytes may thus release glutamate using a mechanism resembling the neuronal vesicular release of neurotransmitters . Synaptobrevin is a vesicular protein that together with plasma membrane proteins syntaxin and SNAP-25 participate in formation of the anchoring core complex required for initiation of exocytosis . Here, we demonstrate that synaptobrevin II is present in cultured astrocytes . Furthermore, we demonstrate that botulinus toxin type B and tetanus toxin cause a decrease in synaptobrevin II immunoreactivity and abolish bradykinin-induced release of glutamate from cultured astrocytes . While we were not able to demonstrate the presence of SNAP-25 or syntaxin immunoreactivity in cultured astrocytes, pretreatment with BoTx-A (which cleaves SNAP-25) and BoTx-C (which cleaves syntaxins) result in a decrease in the baseline release of glutamate and diminish the bradykinin-evoked release of glutamate from cultured astrocytes . These findings strongly support the notion that astrocytes may release neurotransmitters using a mechanism similar to the neuronal secretory process. Am J Orthop, 1997 Mar, 26(3), 201 - 7 Counterparalysis for treatment of paralytic scoliosis with botulinum toxin type A; Nuzzo RM et al.; In this study, botulinum toxin was used to treat paralytic scoliosis . Twelve children with paralytic scoliosis and severe, complicating additional diseases required surgical delay . Although this use of botulinum toxin is experimental, alternative treatments posed greater risks . An institutional review board protocol for nonestablished dosage and indication for treatment was initiated to monitor safety and effect . Treatment was intended to supplement, not replace, other desirable treatment modalities . The effect was to be measured by the return of efficacy of conservative treatment in halting curve progression . Short-term results show that none of the children had worsened scoliosis; all had some reduction in curve measurement (up to >50 degrees). Brain, 1997 Mar, 120 ( Pt 3), 409 - 16 Long-latency reflexes of hand muscles in idiopathic focal dystonia and their modification by botulinum toxin; Naumann M et al.; Long-latency reflexes (LLR) in thenar muscles were elicited by electrical median nerve stimulation in 34 patients with idiopathic focal dystonia and 20 healthy control subjects . Twenty-seven patients had cervical dystonia and seven patients had upper limb dystonia . In about one-quarter of all patients the early LLR (LLR 1, occurring at approximately 40 ms) was abnormal with either increased amplitudes or only unilateral occurrence, mostly on the clinically affected side . Later responses (LLR 2, occurring at approximately 50 ms) were obtained bilaterally in all controls but were reduced or absent in some patients, mostly on the clinically affected side . In 12 dystonia patients, LLR studies were also performed after clinically effective injection of botulinum toxin . Following botulinum toxin treatment there was a significant reduction of LLR 2 amplitudes on the clinically affected side . Our findings suggest a differential involvement of LLR generators in idiopathic dystonia with an antagonism between LLR 1 and LLR 2 on the affected sides . We propose that the reduction of the LLR 2 response may arise from overactivity of the supplementary motor area, confirming the current concept that dystonia results from cortical overflow due to disinhibited thalamocortical pathways projecting to the supplementary motor area . In addition, the dystonic motor pattern seems open to afferent modifications induced by peripheral botulinum toxin treatment. Otolaryngol Head Neck Surg, 1997 Mar, 116(3), 328 - 30 Use of botulinum toxin for diagnosis and management of cricopharyngeal achalasia; Blitzer A et al.; Cricopharyngeal achalasia produces dysphagia in many patients . The spasm has been treated in the past with medication, bouginage, pharyngeal plexus neurectomy, and cricopharyngeal myotomy . In some patients the cause of dysphagia may be elusive, or the patient may be a poor risk for a surgical intervention . We have found that cricopharyngeal muscle injections of botulinum toxin can be useful in the diagnosis and therapy of cricopharyngeal spasm. Dev Med Child Neurol, 1997 Mar, 39(3), 185 - 93 Botulinum toxin A in the hemiplegic upper limb: a double-blind trial; Corry IS et al.; In a randomised, double-blind study, the effects of intramuscular injection of botulinum toxin type A (BtA) into the upper limb were compared with those of normal saline solution in 14 patients with cerebral palsy; their mean age was 9 years . Range of movement and function were assessed before injection and at 2 and 12 weeks after injection . BtA injection significantly increased maximum active elbow and thumb extension and significantly reduced tone at wrist and elbow . The hand grasp-and-release score improved, representing a modest functional change, but fine motor function, assessed by the ability to pick up coins, did not improve and in some cases deteriorated temporarily . The most notable subjective change was the cosmetic benefit of reduced involuntary elbow flexion . The tone-reducing effect of BtA was clinically detectable in comparison with the placebo and patients and parents perceived the change as beneficial . The median of changes in the treatment group was small but the range was large, suggesting that BtA can be useful in selected patients. Naunyn Schmiedebergs Arch Pharmacol, 1997 Mar, 355(3), 335 - 40 Botulinum A toxins: units versus units; Wohlfarth K et al.; We investigated the efficacies and potencies of two commercial preparations of botulinum neurotoxin type A (BoNt/A) reputed to differ in potency . Tests w