• BMB Reports
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• v.30 no.2
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• pp.144-149
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• 1997

An expression vector was constructed containing the gene encoding diphtheria toxin A (DTA) which was placed after a T7 promoter. Cytoplasmic expression of the DTA gene resulted in the formation of an insoluble inclusion body. The inclusion body was collected after the complete lysis of the cell, and subsequent washing with 0.1% Triton X-100 released 16~30% of DTA protein from the inclusion body along with other contaminating proteins. The released DTA protein was purified by dialysis. The remaining pellet was dissolved in 8 M urea containing 5% ${\beta}-mercaptoethanol$, and the denatured DTA was renatured by the dilution-dialysis method. The total yield was 35%, and about 5 mg DTA was obtained from 1 L culture. The DTA protein has a free sulfhydryl group exposed to the protein surface, and was shown to have a tendency to dimerize through disulfide formation in the absence of ${\beta}-mercaptoethanol$. The utility of the sulfhydryl group was tested for the construction of recombinant toxins.

• Journal of Microbiology and Biotechnology
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• v.23 no.5
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• pp.731-737
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• 2013

Seventy-four Shiga toxin-producing Escherichia coli (STEC) isolates belonging to the serotype O91:H21 were isolated from 1,643 asymptomatic human carriers in a STEC outbreak at Gwangju in Korea. Although the isolates did not cause any symptoms, all of them produced Shiga toxins 1 (Stx1) and 2 (Stx2). In order to determine why these strains cause no symptoms, we explored the differences in virulence potential between the asymptomatic STEC O91:H21 isolates and symptomatic STEC O91:H21 strains (ATCC 51435 and ATCC 51434). The asymptomatic STEC O91:H21 isolates showed strongly reduced cytopathic effects compared with the symptomatic strains when intact bacterial cells were used as an inoculant. Moreover, we found a reduced adherence phenotype when testing asymptomatic strains on HeLa cells. Real-time quantitative PCR results suggest that transcriptional repression of the genes encoding type-1 fimbriae occurs in the asymptomatic isolates but not in the symptomatic strains.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.515.2-515
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• 1986

B. sphaericus 1593 K-5 synthesize a potent entomicidal toxin against mosquito larvae. B. sphaericus EcoRl DNA fragment carrying the biocidal activity was clonied and expression in E. coli JM83. For the construction of a recombinant plasmid bearing the toxin activity the DNA of B. sphaericus was partially digested by the EcoRl. The EcoRl DNA fragments were ligated to plasmid pUC 8-EcoR1 site. The transformants were selected on LB plates containing X-gall and ampicilline. The transformants were bioassayed against mosquito larvae of which two clones showed biocidal activity. The two clones were redigested with Eco R1 and analyzed by 0.7% agarose gel electrophorsis.

• Archives of Plastic Surgery
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• v.36 no.4
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• pp.469-474
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• 2009

Purpose: A lower facial contouring surgery has become a commonly performed procedure in Asia. Currently, mandibular angle sagittal ostectomy and botulinum toxin type A treatment are main procedures for aesthetic correction of a broad lower face. There are a few date to show the differences in the mandibular contouring changes between mandibular angle splitting ostectomy and botulinum toxin type A treatment. Facial golden mask is easy to apply, inexpensive, and relatively objective for evaluation of facial contour analysis. This study was designed specifically to compare the changes in lower face width after two different forms of lower facial contouring procedure using facial golden mask. Methods: Seventeen patients, aged 18 to 55 years (mean, 28.6 years), 15 women and 2 men, consented to the study and receive a contouring procedure of lower face. The patients were classified in to 2 groups. In group A, the sample consisted of 10 patients with a prominent squared mandibular angle and mandibular angle splitting ostectomy was performed. In group B, the sample consisted of 7 patients with masseteric hypertrophy and botulinum toxin type A treatment was performed. Photographs of the face were taken to record the facial change at preoperative and postoperative. The postoperative photographs were taken to considered maximal effect at 2 years after surgery in group A and 4.8 months after treatment in group B. The authors applied the facial golden mask to preoperative and postoperative photographs and horizontal ratio, which compares facial width with golden mask width, were calculated. We made an analysis of the result of horizontal ratio using SPSS. Results: Overall average horizontal ratio of pre- and postoperative photos of group A were 1.24 and 1.11, whereas overall average horizontal ratio of pre- and postoperative photos of group B were 1.19 and 1.12. The horizontal ratio decreased 10.24% in group A and 5.93% in group B. There was a statistically significant change in before and after treatment, but there was no significant change in comparing the group A and group B. Conclusions: The result from this study suggest that mandibular angle sagittal ostectomy and botulinum toxin type A treatment showed relatively satisfactory clinical effects on lower facial contouring treatment. There was no statistical significant difference within two lower facial contouring treatment. Facial golden mask is easy to apply, inexpensive, and relatively objective, so we think that facial golden mask is a good method for evaluation of lower facial contouring treatment.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.37
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• pp.10.1-10.5
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• 2015

Background: The aim of this study was to investigate the effect of a botulinum toxin type A (BTX-A) injection in the masseter muscle using electromyography (EMG) in an animal model. Methods: Ten male adult (>3 months of age) New Zealand white rabbits were used. Muscle activity was continuously recorded from 8 hours before to 8 hours after BTX-A injection. The rabbits received unilateral BTX-A injections of either 5 units (group 1, n = 5) or 20 units (group 2, n = 5). Results: The masseter muscle activity of the rabbits was significantly reduced immediately after BTX-A injection (P < 0.05 for both groups). When the results from group 1 were compared with those from group 2, only the peak voltage was significantly decreased in group 2 (P = 0.013). Conclusion: Masseter muscle activity measured by EMG was immediately decreased after a BTX-A injection.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.5
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• pp.485-493
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• 1997

We investigated the effect of ${\alpha}-adrenergic$ and cholinergic receptor agonists on $Ca^{2+}$ current in adult rat trigeminal ganglion neurons using whole-cell patch clamp methods. The application of acetylcholine, carbachol, and oxotremorine ($50\;{\mu}M\;each$) produced a rapid and reversible reduction of the $Ca^{2+}$ current by $17{\pm}6%,\;19{\pm}3%,\;and\;18{\pm}4%$, respectively. Atropine, a muscarinic antagonist, blocked carbachol- induced $Ca^{2+}$ current inhibition to $3{\pm}1%$. Norepinephrine ($50\;{\mu}M$) reduced $Ca^{2+}$ current by $18{\pm}2%$, while clonidine ($50\;{\mu}M$), an ${\alpha}2-adrenergic$ receptor agonist, inhibited $Ca^{2+}$ current by only $4{\pm}1%$. Yohimbine, an ${\alpha}2-adrenergic$ receptor antagonist, did not block the inhibitory effect of norepinephrine on $Ca^{2+}$ current, whereas prazosin, an ${\alpha}1-adrenergic$ receptor antagonist, attenuated the inhibitory effect of norepinephrine on $Ca^{2+}$ current to $6{\pm}1%$. This pharmacology contrasts with ${\alpha}2-adrenergic$ receptor modulation of $Ca^{2+}$ channels in rat sympathetic neurons, which is sensitive to clonidine and blocked by yohimbine. Our data suggest that the modulation of voltage dependent $Ca^{2+}$ channel by norepinephrine is mediated via an α1-adrenergic receptor. Pretreatment with pertussis toxin (250 ng/ml) for 16 h greatly reduced norepinephrine- and carbachol-induced $Ca^{2+}$ current inhibition from $17{\pm}3%\;and\;18{\pm}3%\;to\;2{\pm}1%\;and\;2{\pm}1%$, respectively. These results demonstrate that norepinephrine, through an ${\alpha}1-adrenergic$ receptor, and carbachol, through a muscarinic receptor, inhibit $Ca^{2+}$ currents in adult rat trigeminal ganglion neurons via pertussis toxin sensitive GTP-binding proteins.

• Food Science and Biotechnology
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• v.17 no.4
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• pp.761-765
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• 2008

Bacillus cereus causes two different types of food poisoning syndromes: diarrhea and emesis. The diarrheal syndrome is attributed to various enterotoxins, including nonhemolytic enterotoxin, hemolytic enterotoxin, and enterotoxin-T, whereas the emetic syndrome is caused by the dodecadepsipeptide toxin cereulide. A multiplex polymerase chain reaction (PCR) assay was developed to rapidly detect and identify B. cereus strains. Three primer pairs specific to regions within genes encoding nonhemolytic enterotoxin (nheA), molecular chaperonin (groEL), and cereulide synthetase (ces) were used to identify and differentiate between the enterotoxin-producing and emetic toxin-producing B. cereus strains. The cereulide-producing emetic B. cereus showed 3 PCR products of 325, 405, and 685 bp for the groEL, ces, and nheA genes, respectively, whereas the enterotoxin-producing B. cereus showed 2 PCR products without a ces gene specific DNA fragment. Specific amplifications and differentiations by multiplex PCR assay were obtained using 62 B. cereus strains and 13 strains' of other bacterial species. The detection limit of this assay for enterotoxin-producing strain and emetic toxin-producing strain from pure cultures were $2.4{\times}10^1$ and $6.0{\times}10^2\;CFU/tube$, respectively. These results suggest that our multiplex PCR method may be useful for the rapid detection and differentiation of B. cereus strains in foods.

• International Journal of Industrial Entomology and Biomaterials
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• v.8 no.1
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• pp.89-93
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• 2004

Expression of a fusion protein between B. thuringiensis crystal protein, Cry1Ac1 and a scorpion insect toxin (AaIT, Androctonus australis Hector insect toxin) in acrystalliferous B. thuringiensis strain (Cry-B strain) was examined. The cry 1Ac1 gene was cloned in B. thuringiensis-E coli shuttle vector, pHT3101, under the control of the native cry 1Ac1 gene promoter (pProAc) and a gene encoding AaIT was inserted in XhoI site in the middle of the cry 1Ac1 gene (pProAc-ScoR). B. thuringiensis Cry-B strain carrying pProAc-ScoR (PyoAc-ScoR/CB) produced an inclusion body of irregular shape and the expressed fusion protein is approximately 65 kDa in size. Sporulated cells and spore-crystal mixtures of ProAc-ScoR/CB had insecticidal activity against Plutella xylostella larvae, showing $LT_50$ of ProAc-ScoR/CB (22.59 hrs) lower than that of ProAc/CB (30.06 hrs) at $1{\times}{10^7} {CEU/cm^2}$. These results suggest that the fusion protein including a B. thuringiensis crystal protein and an AaIT may be functionally expressed in B. thupingiensis. Moreover, we verified the additive toxicity of AaIT, which is a new feasible candidate for insect control.

• Journal of Dental Rehabilitation and Applied Science
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• v.23 no.2
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• pp.171-178
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• 2007

Botulinum toxin type A (BTX-A) has a local effect at the neuromuscular junction by blocking acetylcholine release and thus causing paralysis and atrophy of the affected muscles. In dentistry, Botulinum toxin type A(BTX-A) is used for the treatment of masseteric hypertrophy, temporomandibular disorder, and severe bruxism related neurologic disorder. We hypothesized that the muscle atrophy after BTX-A injection into masseter muscle in growing rats, could affect the jaw growth. The purpose of this study was to determine the effects of the BTX-A injected into the masseter muscle on the jaw growth in rats. Rats were divided into four groups(group 1; control group, group 2; saline injection group, group 3; BTX-A injection group, group 4; baseline control group). Group 4 was sacrificed at the beginning of the experiment to provide baseline values of jaw measurements. The weight, length and width of jaw in those groups were measured every weeks. This study reported that the mandibular body length, condylar length, coronoid process length, anterior region height, coronoid process height and condylar height of the jaw in BTX-A injection group were shorter than those of the control and saline injection groups(P<0.05). In conclusion, BTX-A injected into the masseter muscle may affect the undergrowth of the jaw in rats.

• Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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• v.11 no.2
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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.