Mouse Units Research Articles

Autonomic Side Effects of Botulinum Toxin Type B Treatment of Cervical Dystonia and Hyperhidrosis

Recently, botulinum toxin type B (BT-B) has become available to treat muscle hyperactivity in cervical dystonia (CD). When we started the clinical use of BT-B, we noticed a side effect profile not seen with botulinum toxin type A (BT-A) before. Altogether 30 consecutive patients were included in this open controlled study. 24 patients were treated for CD with 11,310 ± 2,616 mouse units (MU) of BT-B (NeuroBloc^®) and 6 for focal hyperhidrosis (HH) with 4,000–10,000 MU. In 5 of them, BT-A (Botox^®) was used additionally for comparison of effectiveness. In CD, side effects consisted of dryness of mouth (total 21, duration 4.4 ± 2.0 weeks, 10 severe, 7 moderate, 4 mild), accommodation difficulties (7), conjunctival irritation (5), reduced sweating (4), swallowing difficulties (3), heartburn (3), constipation (3), bladder voiding difficulties (2), head instability (1), dryness of nasal mucosa (1) and thrush (1). In HH, side effects consisted of accommodation difficulties (4), dryness of mouth (2) and conjunctival irritation (1). Autonomic side effects occur far more often after BT-B than after BT-A. Their localization suggests systemic BT-B spread. BT-B should be applied carefully in patients with pre-existent autonomic dysfunction, additional anticholinergic treatment and in conditions where anticholinergics are contraindicated.

Botulinum toxin A in the treatment of spasticity – An open label study

To assess the efficacy of botulinum toxin type A in spasticity in upper-motor neuron syndromes. Twenty-three patients with spasticity resulted from stroke-related hemiplegia, transverse myelitis and multiple sclerosis took part in the study. Following the history and physical examinations of the patients, injections of botulinum toxin-A were applied. The dose ranged from 80 to 400 mouse unit (MU) depending on the size of the muscle injected. In all patients, spasticity, spasms and pain were measured using the Ashworth Scale, Spasm Frequency Score, and Visual Analogue Scale prior to the therapy, at the 1st week, 1st month and 3rd month of the therapy. In all patients, botulinum toxin type A led to a significant decrease in spasticity, spasms and pain after the 1st week, 1st and 3 rd months of the treatment when compared to the baseline values (p<0.001). No significant side effects or complications were observed. Our results have demonstrated that botulinum toxin type A is effective in the management of patients with spasticity due to stroke-related hemiplegia, transverse myelitis and multiple sclerosis, without major adverse effects.

Botulinum toxin A and the cutaneous nociception in humans: A prospective, double-blind, placebo-controlled, randomized study

Aside from temporary chemodenervation of skeletal muscle and potential anti-inflammatory effects, a genuine peripheral antinociceptive effect of Botulinum Neurotoxin Type A (BoNT/A) has been suspected. To evaluate the effect of BoNT/A on cutaneous nociception in humans, 50 healthy volunteers received subcutaneous injections of 100 mouse units (MU) BoNT/A (Dysport®) and placebo. Both forearms of each subject were treated in a double-blind fashion, one with verum, one with placebo. Heat and cold pain thresholds within the treated skin areas were measured with quantitative sensory testing (QST) and pain thresholds were evaluated with local electrical stimulation (ES). The tests were done before treatment, and after 4 and 8 weeks. No major side effects were noted. All participants completed the study. Heat and cold pain thresholds increased from baseline to week 4 by 1.4 °C for verum and by 1.1 °C for placebo. From baseline to week 8, the thresholds increased by 2.7 °C for verum and by 1.2 °C for placebo. Electrically induced pain thresholds shifted from baseline to week 4 by −0.07 mA for verum and by 0.01 mA for placebo. From baseline to week 8, the thresholds increased by 0.10 mA for verum and by 0.11 mA for placebo. None of these differences was statistically significant. The study shows that there is no direct peripheral antinociceptive effect of BoNT/A in humans. The efficacy of BoNT/A in various pain syndromes must be explained by other pathways such as chemodenervation or anti-inflammatory effects.

Tetrodotoxin in Gastropods (Snails) Implicated in Food Poisoning in Northern Taiwan

The toxin in the gastropods (snails) Zeuxis sufflatus and Niotha clathrata implicated in a food poisoning incident in northern Taiwan in April 2001 was studied. The symptoms exhibited by four victims were general paresthesia, paralysis of the phalanges and the extremities, paralysis, coma, vomiting, and aphasia. The remaining gastropods were assayed for toxicity in the form of tetrodotoxin (TTX). The ranges of specimen toxicity were 345 to 1,640 mouse units (MU) for Z. sufflatus and 190 to 643 MU for N. clathrata. The toxicities of the digestive gland and for other parts of the gastropod were 1,120 ± 477 MU and 497 ± 238 MU, respectively, for Z. sufflatus and 683 ± 113 MU and 289 ± 169 MU, respectively, for N. clathrata. The toxin from the methanolic extract of the gastropods was partially purified by ultrafiltration and Bio-Gel P-2 column chromatography. Cellulose acetate membrane electrophoresis, thin-layer chromatography, high-performance liquid chromatography, and gas chromatography–mass spectrometry analyses demonstrated that the toxin consisted of TTX. It was concluded that the causative agent of the food poisoning in question was TTX.

Long-term results and prognostic factors in the treatment of achalasia with botulinum toxin.

Endoscopic therapy of achalasia by injection of botulinum toxin into the lower esophageal sphincter has very limited adverse effects and is initially successful in 70 % of patients. However, this result only lasts for 6 - 9 months on average in most patients and only half of them benefit for more than 1 year. The aim of this study was to find out which factors are predictive for a good long-term success. We retrospectively studied 25 patients with achalasia. The diagnosis had been proven by barium swallow and esophageal manometry. Therapy was carried out endoscopically between June 1996 and December 1998 by injection of 25 mouse units (MU) botulinum toxin into each of the four quadrants of the lower esophageal sphincter (LES). Lower esophageal sphincter pressure (LESP) was measured prior to and 1 week after endoscopic therapy. A standardized questionnaire was used for symptom assessment, at the initial presentation, at 1 week and at 2.5 +/- 0.8 years after treatment. The LESP was significantly reduced (pre-treatment 62.1 +/- 15.2 mmHg vs. post-treatment 43.1 +/- 12.5 mmHg; P < 0.01). Symptoms improved in 16 patients (pre-treatment symptom score 9.5 +/- 2.9 vs. post-treatment symptom score 4.7 +/- 1.8; P < 0.01). Nine patients showed no relevant improvement. From the 16 patients with a good initial response, two were lost to follow-up. In nine patients the outcome was still satisfactory after a mean of 2.5 years (1.5 - 4 years) (pre-treatment symptom score 9.5 +/- 2.9 vs. symptom score at 2.5 years after Botox 5.1 +/- 1.5; P < 0.05). These patients were on average 67.7 +/- 12.5 years old. The remaining five patients received a second or third injection of botulinum toxin, but none improved substantially for more than 6 months. One of them eventually underwent pneumatic dilation, and three laparoscopic myotomy. Thus, botulinum toxin treatment was unsuccessful in 14 patients in all. These 14 patients were, on average, significantly younger than the nine successfully treated patients (46.1 +/- 12.6 years vs. 67.7 +/- 12.5 years; P < 0.01) and had significantly higher LESP values prior to botulinum toxin therapy (72.8 +/- 8.9 mmHg vs. 47.8 +/- 9.2 mmHg; P < 0.01). The long-term success of botulinum toxin injection into the LES in patients with achalasia is highest in elderly patients and in patients with an LESP not exceeding the upper normal level prior to treatment by 50 % or more. On the basis of our results, younger patients (< 55 years) with a severe increase in LESP do not seem to benefit from botulinum toxin injection and pneumatic dilation or myotomy may be more advantageous to them.

Paralytic toxins in Taiwanese crab Xanthias lividus

: Paralytic toxicity was detected by tetrodotoxin (TTX) bioassay in four and five specimens of new toxic xanthid crab Xanthia lividus collected from Lanyu, south-east Taiwan in April 1999 and Hsiaoliuchiu south-west Taiwan in November 1999, respectively. The average toxicity of crab specimens collected from Lanyu and Hsiaoliuchiu was 230 ± 94 (mean ± SD) mouse units (MU) and 241 ± 114 MU, respectively. The toxin was partially purified by YM-1 membrane ultrafiltration and Bio-Gel P-2 column chromatography. Electrophoresis, high-performance liquid chromatography and gas chromatography–mass spectroscopy analyses showed that the crab toxin was composed mainly of tetrodotoxin (83%), as well as a small amount of gonyautoxin 1-4 (17%).

Low-Dose Botulinum Toxin-A Treatment of Cervical Dystonia – A Double-Blind, Randomized Pilot Study

The purpose of the study was to evaluate the clinical efficacy of low-dose botulinum toxin (BTX) treatment of cervical dystonia in a double-blind, randomized pilot study. Thirty-one patients with cervical dystonia and a minimum of 2 previous Dysport^® treatments received either a mean total target dose of 547 ± 113 mouse units (MU) (group A, 500 MU Dysport/ml) or a 4-times-diluted preparation 130 ± 32 MU (group B, 125 MU Dysport/ml). Assessment was made before and 4 weeks after the injection using Toronto Western Spasmodic Torticollis Rating Scales (TWSTRS). Additionally, a self-response scale evaluated the patients’ subjective condition for 12 weeks after injection. TWST rating before and after injection revealed comparable clinical improvement in both groups. The severity of cervical dystonia at study entry had no effects on therapeutic effects of either preparation. Self-rating revealed comparable subjective improvement in both groups; however, 3 patients of group B received reinjections due to insufficient effects of the injection. Our findings suggest that low-dose treatment of cervical dystonia with Dysport may be clinically effective during maintenance therapy, at least for a limited period of time. Long-lasting effects of previous Dysport treatments at conventional doses and possibly improved diffusion of a highly diluted toxin preparation may have contributed to the effects of the low-dose regimen. Our data emphasize the need for further studies in order to clearly identify optimal toxin preparations for therapeutic uses of BTX-A in neurologic disorders.

Are puffer fish more toxic in their spawning seasons?

It has been a common belief that all puffer fishes are more toxic in their spawning seasons, perhaps to protect their fertilized eggs. However, the current study of two puffer fishes, Takifugu niphobles (Jordan and Snyder) and Takifugu alboplumbeus (Richardson), collected in Hong Kong waters continuously for over a year and bioassayed for toxicity in terms of mouse units (MU), showed the contrary. Their annual spawning seasons (evident from the enlargement of the gonads and the presence of eggs in the ovary) were found to be from October to February (4 consecutive months) and December to February (2 consecutive months), respectively. In both species, the ovaries were highly toxic (100–1,000 MU/g) only in their non-spawning seasons and they became weakly toxic (10–100 MU/g) in their spawning seasons; unlike the ovaries, liver and intestine, the flesh and the skin did not have marked annual toxicity patterns. Their flesh was basically non-toxic (less than 10 MU/g) throughout the whole year. The testes, which were only developed in the spawning seasons, were also found to be non-toxic.

The annual toxicological profiles of two common puffer fish, Takifugu niphobles (Jordan and Snyder) and Takifugu alboplumbeus (Richardson), collected along Hong Kong coastal waters

The toxicological profiles of two local common puffer fish, Takifugu niphobles (Jordan and Snyder) and Takifugu alboplumbeus (Richardson), collected in Hong Kong waters were investigated continuously for 14 months (June 1997–August 1998). Their annual spawning seasons (as evident by the enlargement of gonads and presence of eggs in the ovary) were found to be from October to February (four consecutive months) and December to February (two consecutive months), respectively. The toxicities of their internal organs were determined by standard mouse bioassay and expressed in terms of mouse units (MU). The ovary (in both species) and liver (only in T. niphobles) were discovered to be moderately toxic (100–1000 MU/g) in their non-spawning seasons. Whilst all the intestine and skin were weakly toxic (between 10 and 100 MU/g), their flesh, however, was basically non-toxic (less than 10 MU/g) throughout the whole year. The testes, which were only developed in their spawning seasons, were also non-toxic. This study showed that the toxicological profiles of both of the two species had prominent seasonal patterns which were highly related to their spawning seasons. Contrary to the most common belief, both species were discovered to be relatively less toxic during their spawning seasons.

Dose thresholds and duration of the local anhidrotic effect of botulinum toxin injections: measured by sudometry.

Local injections of botulinum toxin type A (BTX-A) have been used successfully to treat focal hyperhidrosis, but because experimental data were lacking, doses have been chosen arbitrarily or empirically. To analyse dose dependency and duration of BTX-A-derived suppression of sweat gland activity. Employing a standardized scheme (four injections, square 2 x 2 cm), different doses of BTX-A [Dysport(R); 2.5-120 mouse units (MU)] were injected subcutaneously at the lateral aspects of both of the lower legs in 15 healthy volunteers. Sweat tests were performed before, and 3 weeks and 6 months after, BTX-A injections. Sweating was visualized by staining with iodine starch, and quantified by capacitance hygrometry after carbachol iontophoresis, the quantitative sudomotor axon reflex test (QSART). Iodine starch staining indicated a threshold dose of 10 MU (2.5 MU cm-2) leading to visible anhidrotic skin spots after 3 weeks in all subjects. This was maintained for 6 months with doses of 50 MU (12.5 MU cm-2) or higher, but the size of the anhidrotic skin area decreased over time (P < 0.001) indicating partial recovery at the edges. After 3 weeks QSART was significantly reduced (P < 0.02) and completely suppressed by doses of 80 MU (20 MU cm-2) or more, although after 6 months QSART increased again to pre-BTX-A levels (P < 0.001). Both methods indicated that the suppression of sweating is dose dependent (QSART: r = -0.70, P < 0.001; iodine starch staining: r = 0.74, P < 0.001). Our findings suggest that BTX-A effectiveness can be quantified by testing sudomotor function. For the first time threshold doses for the suppression of sweating have been defined.

Possible source of tetrodotoxin in the starfish Astropecten scoparius

The seasonal variations of toxicity and stomach contents in toxic starfish Astropecten scoparius were detected. The average highest specimen toxicity, expressed as tetrodotoxin (TTX), was 16,821 mouse units (MU). The toxin was composed of TTX only, except April’s sample containing mainly TTX along with minor paralytic shellfish poisons. The composition in the stomach of less and more toxic starfish was mainly Veremolpa scabra and Umborium suturale, respectively. The toxicities of U. suturale and V. scabra, found in the starfish stomach were 65 and 33 MU, respectively. For those collected from the coastal waters, U. suturale was toxic with average toxicity value of 77 MU/g, but V. scabra was nontoxic. The toxin in the specimens of U. suturale collected from either the digestive gland of starfish or the coastal waters, was TTX and anhydroTTX only. It indicates that the starfish A. scoparius might mainly accumulate high amount of TTX from U. suturale. Furthermore, both small gastropods U. suturale and Natica psuestes are first reported to contain TTX.

Paralytic toxins in taiwanese starfish Astropecten scoparius

Paralytic toxicity was seasonally detected in every 10 specimens of the starfish Astropecten scoparius from Pingtung in Taiwan from December 1995 to November 1996. The highest toxicity and the average toxicity of total specimens, expressed as tetrodotoxin (TTX), were 5,938 and 1,679 mouse unit (MU), respectively. The highest value of monthly average toxicity in viscera and other parts were 354 and 247 MU/g, respectively. The toxin was partially purified by YM-2 membrane ultrafiltration and Bio-Gel P-2 column chromatography. HPLC and GC-MS analyses showed that the starfish toxin was mainly composed of tetrodotoxin (88%), along with minor gonyautoxin 2-3 and saxitoxin (12%).

Anatomical Distribution and Seasonal Variation of Toxicity of Puffer Fish, “Hoshifugu” Arothron firmamentum Specimens Collected from the Bungo Channel, Oita

Forty-nine specimens of “Hoshifugu” puffer fish, Arothron firmamentum (23 male and 26 female specimens) were collected from the Bungo Channel, Oita Prefecture, during 1991 to 1997, and examined for the anatomical distribution and the seasonal variation of toxicity by the mouse assay method.Skin showed a rather low level of toxicity and the frequency of toxic specimens was 6.1% with the maximal lethal potency of 29 mouse unit (MU)/g. Ovary showed a higher level of toxicity (frequency 80.8%) with the maximal lethal potency of 640MU/g. On the other hand, the muscle, liver, testis and intestine were non-toxic. Ovaries collected in 6 different months (March, May, and August-November) were found to be toxic with the frequency of 50-100%. Skins were found to be toxic in August and October. It appears that this species has high toxicity in the ovary during early summer to autumn.Since the muscle of “Hoshifugu” puffer fish specimens caught in the Bungo Channel was non-toxic, the muscle of this species appears to be edible without risk to humans.

Toxicity and toxic components of two xanthid crabs, atergatis floridus and demania reynaudi, in taiwan

Paralytic toxicity was detected by tetrodotoxin bioassay in eight specimens of Atergatis floridus and seven specimens of Demania reynaudi, collected from Taiwan in 1994. The toxicity of crab specimens was 161 ± 115 (mean ± S.D.) mouse units (MU) for A. floridus and 640 ± 273 MU for D. reynaudi. The respective toxins were partially purified from specimens of A. floridus and D. reynaudi by ultrafiltration using a Diaflo YM-1 membrane, followed by chromatography on a Bio-Gel P-2 column. Electrophoresis, thin-layer chromatography, high-performance liquid chromatography, ultraviolet spectrum and gas chromatography—mass spectrometry indicated that the toxin of A. floridus was mainly composed of tetrodotoxin (85%), along with minor gonyautoxin 1—4 (15%) and the toxin of D. reynaudi was mainly composed of tetrodotoxin (88%), along with minor gonyautoxin 2—4 and neosaxitoxin (12%).

Patient selection in the treatment of glabellar wrinkles with botulinum toxin type A injection.

To determine the dose-response characteristics and side-effects profile of Clostridium botulinum type A exotoxin (Botox) used to treat glabellar wrinkles and develop guidelines for patient selection based on the nature and severity of the treated wrinkles. Prospective, nonrandomized pilot and electromyogram (EMG)-guided studies. Two ambulatory care clinics at university hospitals. For the pilot study, volunteer samples of 23 patients with glabellar wrinkles; for the EMG-guided study, volunteer samples of 57 patients with glabellar wrinkles. For the pilot study, 23 patients were serially injected with up to 10.0 mouse units (MU) of Botox into each corrugator muscle; for the EMG-guided study, 57 patients were injected under EMG guidance with an initial dose of 10.0 MU of Botox into each corrugator muscle. Eleven patients with persistent corrugator activity were reinjected with 10.0 MU of Botox. For the pilot study, slide photographs were obtained before and 2 weeks after injection; for the EMG-guided study, slide photographs were obtained before and at 2 weeks and at 2 months after injection. Patients were asked to evaluate results numerically. For the pilot study, injection of up to 10.0 MU of Botox into each corrugator muscle produced a satisfactory improvement in 12 patients; for the EMG-guided study, 43 patients were satisfied with improvement after full abolition of corrugator or accessory lateral brow muscle activity. Women were more likely to achieve satisfactory results than were men (80% [40/50] vs 43% [3/7]; P < or = .03). Improvement was not age related. No significant side effects or complications were observed. Glabellar wrinkles may be satisfactorily treated with Botox injection into the corrugator supercilii muscles. Improvement is temporary, dose dependent, and may not be seen in some patients even with successful denervation of the treated muscles. Clinicians may begin treatment with a dose of 10.0 MU of Botox into each corrugator muscle, and may select candidates for injection by determining the type of wrinkle to be treated and its spreadability (glabellar spread test).

A Preliminary Study of Puffer Fishes and Their Toxins Found in Hong Kong Waters

Puffer fishes are commonly found in Hong Kong waters. For this study, we collected and identified six species of local puffers, Takifugu niphobles (Jordan and Snyder), Takifugu alboplumbeus (Richardson), Chelonodon patoca (Hamilton-Buchanan), Lagocephalus spadiceus (Richardson), Takifugu ocellatus (Osbeck) and Diodon holocanthus (Linnaeus). The first three species were confirmed to be toxic (6-120 mouse units (MU)/g) by mouse bioassay. To isolate tetrodotoxin (TTX) from a common local puffer, Takifugu niphobles, two hundred grams of viscera (ovary, intestine and liver) was minced and extracted with acidic methanol. The crude toxin obtained was further purified by Sephadex G-50-50 gel filtration and Whatman CM-52 cation exchange column chromatography. Purification afforded two unidentified toxins, unknown 1 and unknown 2, which gave single fluorescent spots under UV irradiation (366nm) with Rf values of 0.80 and 0.54, respectively, in thin layer chromatography (TLC). The electrospray ionization mass spectra (ESI/MS) of unknowns 1 and 2 had their molecular ion peaks (M+H)+ at m/z 320 and 255, respectively. Unknown 1 was concluded to be TTX, whereas unknown 2 was probably a derivative of TTX, whose exact identification would require further chemical analyses.

Use of botulinum toxin in stroke patients with severe upper limb spasticity.

Spasticity can contribute to poor recovery of upper limb function after stroke. This is a preliminary evaluation of the impact of botulinum toxin treatment on disability caused by upper limb spasticity after stroke. Seventeen patients with severe spasticity and a non-functioning arm were treated with intramuscular botulinum A neurotoxin (median age at treatment 54.5 years; median time between onset of stroke and treatment 1.5 years). Baseline and assessments two weeks after treatment were compared to assess efficacy. The duration of improvement in disability was documented. Outcome measures used were; passive range of movement at the shoulder, elbow, wrist, and fingers; modified Ashworth scale to assess spasticity of biceps and forearm finger flexors; an eight point scale to assess the degree of difficulty experienced by the patient or carer for each functional problem defined before treatment; the presence of upper limb pain. The biceps, forearm finger flexors, and flexor carpiulnaris were treated with intramuscular botulinum toxin. Up to a total dose of 400-1000 mouse units (MU) of Dysport (Speywood) or 100-200 MU of BOTOX (Allergan) was used in each patient. Functional problems reported by the patients before treatment were difficulty with cleaning the palm, cutting fingernails, putting the arm through a sleeve, standing and walking balance, putting on gloves, and rolling over in bed. Hand hygiene improved in 14 of 17 patients; difficulty with sleeves improved in four of 16; standing and walking balance improved in one of four; shoulder pain improved in six of nine; wrist pain improved in five of six. Passive range of movement at shoulder, elbow, and wrist improved after treatment. Benefit was noted within two weeks and lasted one to 11 months. No adverse effects occurred. This preliminary study suggests that intramuscular botulinum toxin is a safe and effective treatment for reducing disability in patients with severe upper limb spasticity.

Disability Changes After Treatment of Upper Limb Spasticity with Botulinum Toxin

We performed an open label single-blind trial with seventeen patients with moderate to severe upper limb spasticity following stroke or traumatic brain injury, treated with an average dose of 165 MU (mouse units) botulinum toxin (range: 100 MU to 210 MU). Evaluation of outcome was carried out analyzing the changes in standard mea sures of impairment, disability, and quality of life. Spasticity reduction, with increased range of motion (ROM), was observed in all patients, independent of the duration of spasticity. Changes either in focal or in global disability scores occurred in two sub groups of subjects showing, respectively, residual motor function of the extensor mus cles or severe spasticity-related disability. Patients with no residual motricity of the antagonist muscles at the spastic limb, with moderate spasticity, scoring high values on the Functional Independence Measure (FIM) assessment, did not show any disabil ity decrease after treatment. These findings stress the need for an accurate as...

Paralytic Shellfish Poisons of Ormer, Haliotis tuberculata, from Spain

Paralytic toxicity in excess of the quarantine limit of 4 mouse unit (MU)/g as paralytic shellfish poisons (PSPs) was detected in ormer, Haliotis tuberoulata, imported from Vigo, Spain, to Japan, between January and April, 1994. The ormer exhibited an unprecedented anatomical distribution of PSP toxicity. The muscular tissues were highly toxic, in comparison with the visceral ones. Most toxic was the foot (highest toxicity, 106MU/g), followed by the epipodium of the foot (30.9MU/g), mouth (33.2MU/g), viscera (15.6MU/g), right shell muscle (8.2MU/g), and digestive gland (3.7MU/g). The toxicity scores of ctenidium markedly varied in specimens (6.4-99.9MU/g).PSPs were extracted from toxic specimens of the ormer with 80% ethanol acidified to pH 3-3.5, and partially purified by chromatography on a Sep-pak C18 cartridge column and ultrafiltration through an Ultrafree CL-LCC. HPLC analysis demonstrated that the ormer toxins apparently comprised members of the saxitoxin (STX) group, such as STX, neoSTX, and decarbamoylSTX (dcSTX). Regardless of the tissue, dcSTX was the major PSP component, which accounted for 83mol% (epipodium of foot) to 97mol% (digestive gland) of all toxin components. Electrospray ionization mass spectral analysis confirmed that the principal toxin component from the Spanish ormer was dcSTX. Neither gonyautoxin group nor C toxins (N-sulfocarbamoyl-11-hydroxysulfate PSP derivatives) appeared in any tissues. No significant individual variation in PSP composition was observed. These results reveal toxification with PSPs of a herbivorous marine gastropod, and suggest a unique metabolism of PSPs in the ormer.

Crystalline Preparation of Botulinum Toxin Type a (Botox): Degradation in Potency with Storage

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