• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.1
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• pp.295-301
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• 2017

Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.4
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• pp.349-355
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• 2015

We examined the effects of peripherally or centrally administered botulinum neurotoxin type A (BoNT-A) on orofacial inflammatory pain to evaluate the antinociceptive effect of BoNT-A and its underlying mechanisms. The experiments were carried out on male Sprague-Dawley rats. Subcutaneous (3 U/kg) or intracisternal (0.3 or 1 U/kg) administration of BoNT-A significantly inhibited the formalin-induced nociceptive response in the second phase. Both subcutaneous (1 or 3 U/kg) and intracisternal (0.3 or 1 U/kg) injection of BoNT-A increased the latency of head withdrawal response in the complete Freund's adjuvant (CFA)-treated rats. Intracisternal administration of N-methyl-D-aspartate (NMDA) evoked nociceptive behavior via the activation of trigeminal neurons, which was attenuated by the subcutaneous or intracisternal injection of BoNT-A. Intracisternal injection of NMDA up-regulated c-Fos expression in the trigeminal neurons of the medullary dorsal horn. Subcutaneous (3 U/kg) or intracisternal (1 U/kg) administration of BoNT-A significantly reduced the number of c-Fos immunoreactive neurons in the NMDA-treated rats. These results suggest that the central antinociceptive effects the peripherally or centrally administered BoNT-A are mediated by transcytosed BoNT-A or direct inhibition of trigeminal neurons. Our data suggest that central targets of BoNT-A might provide a new therapeutic tool for the treatment of orofacial chronic pain conditions.

• The Korean Journal of Pain
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• v.18 no.1
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• pp.29-33
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• 2005

Background: Chronic headache (CH) constitutes a significant public health problem, impacting on both the individual sufferer and society. Patients with CH, unresponsive to drug therapy or nerve block, suffer considerable disability due to the frequency and severity of attacks; therefore, they should be considered for novel therapy. Botulinum toxin type A (BoNT-A) has shown significant promise in the management of CH. In this paper, we review recent evidence on the efficacy of BoNT-A, and also report our experience with this treatment in CH patients. Methods: BoNT-A was used to treat 69 CH patients, including 47 in a chronic migraine group and 22 in a non-migraine CH group, who showed therapy-resistance to palliative drug or nerve block. We investigated the demography, dosage and site of BoNT-A injection, and used a visual analogue scale (VAS) for pain and the degree of satisfaction. The data were analyzed using t-tests and a Friedman repeated measures analysis of variance on ranks. Results: Significant decreases in the VAS for pain were found in both the chronic migraine and non-migraine CH groups, from 2, 4 and 12 weeks and from 4 and 12 weeks, respectively, after BoNT-A administration (P < 0.05). The chronic migraine group showed significantly lower VAS scores for pain than the non-migraine CH group from 2, 4 and 12 weeks after the BoNT-A administration (P < 0.05). Twenty eight patients (59.2%) in the chronic migraine group and eight (36.4%) in the non-migraine CH were satisfied with the BoNT-A treatment. Conclusions: This clinical study revealed that the use of BoNT-A demonstrated efficacy for CH patients resistant to drug therapy or nerve block. Moreover, BoNT-A proved itself more effective in the chronic migraine than non-migraine CH group.

• Journal of Veterinary Clinics
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• v.28 no.3
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• pp.273-279
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• 2011

Botulinum toxin type A (BoNT/A) has been proven to be a safe and effective treatment for wrinkles in human. As well as the anti-wrinkle effects, the anti-pruritic effect of BoNT/A has been revealed from several researches for new therapy. The aim of this study was to investigate the anti-pruritic effect of BoNT/A against histamine-induced pruritus on canine skin. Five clinically healthy beagles were included in the study. All dogs were received 0.05 ml (5 Unit) of BoNT/A on the right dorsal thoracic region as an experiment and the same volume of saline solution was injected on the left dorsal thoracic region as a control, respectively. Intradermal histamine injections were performed four times (before treatment and days 1, 3 and 7 after BoNT/A injection). The severity of pruritus, the diameter and thickness of wheal, the erythema index and cutaneous temperature were assessed. The severity of pruritus was reduced on BoNT/A treated sides, compared with saline treated sides (p < 0.05). BoNT/A decreased wheal size, in both diameter and thickness (p < 0.05). Although, erythema index of both sides were increased after first histamine injection, BoNT/A treated sides showed the low-value as compared with saline treated sides. Cutaneous temperature was decreased significantly on BoNT/A treated sides. These results indicates that BoNT/A reduce histamine-induced pruritus on canine skin and suggested a possibility of application of BoNT/A for local intractable dermatologic problem in dogs.

• Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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• v.30 no.2
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• pp.77-81
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• 2019

Dysphagia may result from dysfunction of any of the components involved in the complex neuromuscular interaction of swallowing. Hyperfunction of any of the muscles involved in swallowing is a frequent cause of dysphagia. The cricopharyngeus muscle (CPM) is a key component of the upper esophageal sphincter. Cricopharyngeus muscle dysfunction (CPD) refers to the muscle's failure to appropriately and completely relax or expand during deglutition. A variety of disease processes may cause CPD and accurate diagnosis is paramount for appropriate treatment. In appropriately selected patients, intervention at the CPM may yield significant improvement in dysphagia. Interventions include nonsurgical, pharyngoesophageal segment dilatation, botulinum toxin (BoNT) injection, and criccopharyngeal myotomy. Injections of BoNT in patients with CPD have been reported to result in marked relief of dysphagia. Different techniques for instilling BoNT into the CPM have been described. Awake, in-office CPM BoNT injection with electromyography and/or fluoroscopic or ultrasound guidance is performed transcervically or via flexible endoscopy. Operative CPM BoNT injection involves rigid laryngoscopy and esophagoscopy with direct visualization of the CPM. BoNT should be prepared in low-volume, high-concentration dilutions to minimize the potential for undesired diffusion of the toxin. The effects of BoNT occur within weeks of injection and typically last up to 5 or 6 months.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.5
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• pp.539-546
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• 2018

Botulinum toxin type A (BoNT/A) has been used therapeutically for various conditions including dystonia, cerebral palsy, wrinkle, hyperhidrosis and pain control. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) receive orofacial nociceptive information from primary afferents and transmit the information to higher brain center. Although many studies have shown the analgesic effects of BoNT/A, the effects of BoNT/A at the central nervous system and the action mechanism are not well understood. Therefore, the effects of BoNT/A on the spontaneous postsynaptic currents (sPSCs) in the SG neurons were investigated. In whole cell voltage clamp mode, the frequency of sPSCs was increased in 18 (37.5%) neurons, decreased in 5 (10.4%) neurons and not affected in 25 (52.1%) of 48 neurons tested by BoNT/A (3 nM). Similar proportions of frequency variation of sPSCs were observed in 1 and 10 nM BoNT/A and no significant differences were observed in the relative mean frequencies of sPSCs among 1-10 nM BoNT/A. BoNT/A-induced frequency increase of sPSCs was not affected by pretreated tetrodotoxin ($0.5{\mu}M$). In addition, the frequency of sIPSCs in the presence of CNQX ($10{\mu}M$) and AP5 ($20{\mu}M$) was increased in 10 (53%) neurons, decreased in 1 (5%) neuron and not affected in 8 (42%) of 19 neurons tested by BoNT/A (3 nM). These results demonstrate that BoNT/A increases the frequency of sIPSCs on SG neurons of the Vc at least partly and can provide an evidence for rapid action of BoNT/A at the central nervous system.

• Archives of Craniofacial Surgery
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• v.22 no.1
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• pp.1-10
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• 2021

Botulinum toxin type A (BoNT-A), onabotulinumtoxinA (Botox) was approved by the United States Food and Drug Administration for temporary improvement of glabellar lines in patients 65 years and younger in 2002, and has also been used widely for aesthetic purposes such as hyperhidrosis, body shape contouring, and other noninvasive facial procedures. BoNT-A inhibits presynaptic exocytosis of acetylcholine (ACh)-containing vesicles into the neuromuscular junction at cholinergic nerve endings of the peripheral nervous system, thereby paralyzing skeletal muscles. ACh is the most broadly used neurotransmitter in the somatic nervous system, preganglionic and postganglionic fibers of parasympathetic nerves, and preganglionic fibers or postganglionic sudomotor nerves of sympathetic nerves. The scientific basis for using BoNT-A in various cosmetic procedures is that its function goes beyond the dual role of muscle paralysis and neuromodulation by inhibiting the secretion of ACh. Although the major target organs for aesthetic procedures are facial expression muscles, skeletal body muscles, salivary glands, and sweat glands, which are innervated by the somatic or autonomic nerves of the peripheral cholinergic nerve system, few studies have attempted to directly explain the anatomy of the areas targeted for injection by addressing the neural physiology and rationale for specific aesthetic applications of BoNT-A therapy. In this article, we classify the various cosmetic uses of BoNT-A according to the relevant component of the peripheral nervous system, and describe scientific theories regarding the anatomy and physiology of the cholinergic nervous system. We also review critical physiological factors and conditions influencing the efficacy of BoNT-A for the rational aesthetic use of BoNT-A. We hope that this comprehensive review helps promote management policies to support long-term, safe, successful practice. Furthermore, based on this, we look forward to developing and expanding new advanced indications for the aesthetic use of BoNT-A in the future.

• The Korean Journal of Pain
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• v.24 no.3
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• pp.164-168
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• 2011

Obturator nerve block has been commonly used for pain management to prevent involuntary reflex of the adductor thigh muscles. One of several options for this block is chemical neurolysis. Neurolysis is done with chemical agents. Chemical agents used in the neurolysis of the obturator nerve have been alcohol, phenol, and botulinum toxin. In the current case, a patient with spasticity of the adductor thigh muscle due to cervical cord injury had obturator nerve neurolysis done with botulinum toxin type B (BoNT-B). Most of the previous studies have used BoNT-A with only a few reports that have used BoNT-B. BoNT-B has several advantages and disadvantages over BoNT-A. Thus, we report herein a patient who successfully received obturator nerve neurolysis using BoNT-B to treat adductor thigh muscle spasm.

• BMB Reports
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• v.39 no.5
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• pp.642-647
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• 2006

Botulinum neurotoxin A (BoNT/A) has been used therapeutically to treat muscular hypercontractions and sudomotor hyperactivity and it has been reported that BoNT/A might have analgesic properties in headache. PEP-1 peptide is a known carrier peptide that delivers fulll-ength native proteins in vitro and in vivo. In this study, a BoNT/A gene were fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-BoNT/A fusion protein. The expressed and purified PEP-1-BoNT/A fusion proteins were efficiently transduced into cells in a time- and dose-dependent manner when added exogenously in a culture medium. In addition, immuno-histochemical analysis revealed that PEP-1-BoNT/A fusion protein efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin. These results suggest that PEP-1-BoNT/A fusion protein provide an efficient strategy for therapeutic delivery in various human diseases related to this protein.

• Journal of Microbiology and Biotechnology
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• v.29 no.7
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• pp.1165-1176
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• 2019

Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are the most toxic substances known. However, the number of currently approved medical countermeasures for these toxins is very limited. Therefore, studies on therapeutic antitoxins are essential to prepare for toxin-related emergencies. Currently, more than 10,000 Halla horses, a crossbreed between the native Jeju and Thoroughbred horses, are being raised in Jeju Island of Korea. They can be used for equine antitoxin experiments and production of hyperimmune serum against BoNT/A1. Instead of the inactivated BoNT/A1 toxoid, Halla horse was immunized with the receptor-binding domain present in the C-terminus of heavy chain of BoNT/A1 (BoNT/A1-HCR) expressed in Escherichia coli. The anti-BoNT/A1-HCR antibody titer increased rapidly by week 4, and this level was maintained for several weeks after boosting immunization. Notably, $20{\mu}l$ of the week-24 BoNT/A1-HCR(-immunized) equine serum showed an in vitro neutralizing activity of over 8 international units (IU) of a reference equine antitoxin. Furthermore, $20{\mu}l$ of equine serum and $100{\mu}g$ of purified equine $F(ab^{\prime})_2$ showed 100% neutralization of 10,000 $LD_{50}$ in vivo. The results of this study shall contribute towards optimizing antitoxin production for BoNT/A1, which is essential for emergency preparedness and response.