• Molecules and Cells
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• 제24권3호
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• pp.329-337
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• 2007

An enrichment semi-nested PCR procedure was developed for detection of Clostridium botulinum types A, B, E, and F. It was applied to sediment samples to examine the prevalence of C. botulinum in the Korean environment. The first pair of primers for the semi-nested PCR was designed using a region shared by the types A, B, E, and F neurotoxin gene sequences, and the second round employed four nested primers complementary to the BoNT/A, /B, /E, and /F encoding genes for simultaneous detection of the four serotypes. Positive results were obtained from the PCR analysis of five of 44 sediments (11%) collected from Yeong-am Lake in Korea; all were identified as deriving from type B neurotoxin (bontb) genes. Two of the C. botulinum type B organisms were isolated, and their bontb genes sequenced. The deduced amino acid sequences of BoNT/B showed 99.5 and 99.8% identity with the amino acid sequence of accession no. AB084152. Our data suggest that semi-nested PCR is a useful tool for detecting C. botulinum in sediments, and renders it practicable to conduct environmental surveys.

• 대한후두음성언어의학회지
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• 제11권2호
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• pp.185-187
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• 2000

The etiology of vocal fold granuloma was identified : post-endotracheal intubation, vocal abuse, acid reflux and idiopathic. The identification of the cause or causal factor is important, since the treatment must be fundamental directed at them. Treatment have included voice therapy and antireflux measures. Surgical excision is considered in patients who do not respond to medical management. In this study, a case of vocal fold granuloma resolved who underwent injection of the affected vocal fold. Botulinum toxin type A is probably successful by decreasing the strength during adduction in the arytenoid region which, when very intense, would perpetuate the granuloma. Localized injection of this neurotoxin is promising both as an initial treatment and as an alternative treatment in patients who do not respond to standard therapy.

• Journal of Microbiology and Biotechnology
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• 제21권7호
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• pp.719-727
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• 2011

Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

• Journal of Oral Medicine and Pain
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• 제38권4호
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• pp.325-331
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• 2013

보툴리눔 톡신은 신경독소로, 운동신경 말단부위에서 분비되는 아세틸콜린의 분비를 차단하여 근육의 위축을 유발하게 된다. 의학계 및 치의학계에서는 이를 이용하여 다양한 질환을 치료하는 것을 시도하고 있다. 치과영역에서는 저작근 수축, 심한 이갈이, 안면 틱, 구강안면 운동장애, 교근비대의 치료 등 과활성 근육성 질환을 치료하는 데 사용하고 있다. 악안면 영역에 보툴리눔 톡신을 주입하고 난 뒤 발생되고 있는 합병증으로는, 자연스럽지 못한 안면표정, 통증의 증가, 두통 등이 유발될 수 있다고 보고되고 있다. 본 증례에서는 교근부에 보툴리눔 톡신 주입 후 발생된 전방 개교합 증상에 대하여 보고하고자 한다.

• 대한치과의사협회지
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• 제48권12호
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• pp.889-892
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• 2010

Botulinum toxin type A (BTX-A), a potent neurotoxin that reversibly blocks presynaptic acetylcholine release, has been applied successfully to treat facial spastic conditions such as blepharospasm, strabismus and cervical dystonia. Since the first reported application in dentistry in 1994, BTX-A has been used with great success to used in the orofacial region to help treat masticatory and facial muscle spasm, severe bruxism, facial tics, and hypertrophy of the masticatory muscles. The clinician may be aware of the many courses becoming available and aimed at dentists to start using it in the cosmetic context. This article intends to provide a basic understanding of the many functional uses of the drug in the orofacial region that may be relevant to everyday practice, especially in orthodontic field.

• 대한후두음성언어의학회지
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• 제23권2호
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• pp.111-118
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• 2012

Botulinum toxin is a potent neurotoxin that is produced by the bacterium Clostridium botulinum. The agent causes muscle paralysis by preventing the release of acetylcholine at the neuromuscular junction of striated muscle. Botulinum toxin A (Botox, AllerganInc., Irvine, California) is the most potent of seven distinct toxin subtypes that are produced by the bacterium. The toxin was initially used clinically in the treatment of strabismus caused by hypertonicity of the extraocular muscles and was sub-sequently described in the treatment of multiple disorders of muscular spasticity and dystonia. In treating patients with Botox for blepharospasm, Carruthers and Carruthers [5] noticed an improvement in glabellar rhytids. This ultimately led to the introduction and development of Botox as a mainstay in the treatment of hyperfunctional facial lines in the upper face. Since its approval by the U.S. Food and Drug Administration for the treatment of facial rhytids (2002), botulinum toxin A has expanded into wide-spread clinical use. Forehead, glabellar, and periocular rhytids are the most frequently treated facial regions. Indications for alternative uses for Botox in facial plastic and reconstructive surgery are expanding. These include a variety of well-established procedures that use Botox as an adjunctive agent to enhance results. In addition, Botox injection is finding increased usefulness as an independent modality for facial rejuvenation and rehabilitation. The agent is used beyond its role in facial rhytids as an effective agent in the management of dynamic disorders of the face and neck. Botox injection allows the physician to precisely manipulate the balance between complex and conflicting muscular interactions, thus resetting their equilibrium state and exerting a clinical effect. This article will address some of the new and unique indications on Botox injection in the face (the lower face and neck, combination with fillers). Important points in terms of its clinical relevance will be stressed, such as an understanding of functional facial anatomy, the importance of precise injections, and correct dosing all are critical to obtaining natural outcomes.

• 대한치과의사협회지
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• 제55권5호
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• pp.365-369
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• 2017

In clinical dentistry, botulinum toxin is generally used to treat the square jaw, bruxism, and temporomandibular joint diseases. Recently, this procedure has been expanded and applied for cosmetic purposes, and it is becoming a key task to be aware of the precise anatomical structure of the target muscles to be cautious during treatment and how to prevent side effects. Therefore, the purpose of this study is to observe the anatomical structure of the superficial layer of masseter muscle and to provide a most effective botulinum toxin injection method through clinical anatomical consideration. It was observed that the muscle belly of superficial part of the superficial layer was originated from the deep to the aponeurosis of masseter muscle and descend, then changed gradually into the tendon structure attaching to the inferior border of the mandible. In this study, we named this structure deep inferior tendon. This structure was observed in all specimens. We conclude that the use of superficial layer and deep layer injection should be considered to prevent paradoxical masseteric bulging in consideration of the deep inferior tendon of superficial part of superficial layer of masseter muscle.

• 미생물학회지
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• 제13권2호
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• pp.71-80
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• 1975

The neurotoxin of Clostridium botulinum type B was purified from a liquid culture. The purification steps consist of ammonium sulfate precipitation of whole culture, treatment of Polymin P(0.15%, v/v), gel filtration on Sephadex G-100 at pH5.6 and DEAE-Sephadex charomatography at pH8.0. The procedure recovered 17% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis and had a molecular weight of 163,000. Subunits of 106,000 and 56,000 molecular weight were found when purified toxin was treated with a disulfide-reducing agent and electro phoresed on SDS-polyacrylamide gels.

• The Korean Journal of Physiology and Pharmacology
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• 제19권4호
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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.

• Journal of Microbiology and Biotechnology
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• 제29권7호
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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.