No Abstract Available.
Clostridium botulinum type B* ; Clostridium botulinum* ; Clostridium*

No Abstract Available.
Clostridium botulinum type B* ; Clostridium botulinum* ; Clostridium*

Botulism is a rare neuroparalytic disease caused by neurotoxins of Clostridium species. A ten-year-old girl and her mother were admitted to a hospital with symptoms of progressive dizziness, blurred vision, slurred speech, constipation and difficulty in swallowing. These characteristic manifestations and clinical course prompted an examination of the possibility of botulism. Mouse bioassay performed with mother's stool demonstrated type A botulinum toxin and culture of the mother's stool was positive for Clostridium botulinum type A. This is the first case of botulism in Korea.
Animals ; Biological Assay ; Botulinum Toxins ; Botulism* ; Clostridium ; Clostridium botulinum ; Clostridium botulinum type A ; Constipation ; Deglutition ; Dizziness ; Female ; Humans ; Korea* ; Mice ; Mothers ; Neurotoxins

OBJECTIVETo establish the genotype-specific targets plasmids and engineered E.coli strains of botulinum neurotoxins (BoNT) types B and E based on reverse genetics.

METHODSThe gene sequences of BoNT were obtained from GenBank and analyzed using DNAMAN, Lasergene, Vector NTI and BLAST. Two target fragments of BoNT/B and BoNT/E were anchored and then synthesized as 5 and 10 short DNA single strands, respectively. The full-length target sequences were amplified by overlapping PCR and subcloned into pMD 18-T vector, and the recombinant plasmids were identified by restriction enzyme digestion and sequencing.

RESULTSSixty full-length sequences of 4 types of BoNT, namely types A, B, E, and F, were available in GenBank. Two target fragments, BoNT/B of 215 bp and BoNT/E of 360 bp, and their specific primer pairs were anchored after sequence analysis. pMD 18-T-BoNT/B and pMD 18-T-BoNT/E containing these two target sequences were confirmed.

CONCLUSIONThe engineered plasmids and E.coli stains containing the genotype-specific target fragments of BoNT/B and BoNT/E have been constructed successfully.

Base Sequence ; Botulinum Toxins ; genetics ; Botulinum Toxins, Type A ; Clostridium botulinum ; genetics ; isolation & purification ; Gene Targeting ; Genotype ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Botulinum toxin type A is a neurotoxin produced by clostridium botulinum that blocks the presynaptic release of acetylcholine at the neuromuscular junction. This blockade of the neuromuscular junction is definitive, but the existence of nerve sprouting explains the reversible nature of the paralysis induced by injection of this toxin. The clinical effect appears between the 3rd and 7th day after injection. The author had experienced 218 cases from January 1999 to September 1999. We injected toxin on forehead (96 cases), glabella (91 cases), crow's feet (88 cases), neck (12 cases), nasolabial fold (9 cases), masseter muscle (8 cases), and chin (7 cases). The author obtained wrinkles decreased significantly. The effect during the period of activity of the toxin, which lasted 3 to 6 months after injection and masseter muscle area lasted 6 to 9 months. The advantage of botulinum toxin injection is the effective treatment with wrinkles in upper half of face and neck and no disturbance in daily life. The disadvantage of botulinum toxin injection is the feeling of masked face after injection, short duration, and sometimes unwilling facial expression.
Acetylcholine ; Botulinum Toxins* ; Botulinum Toxins, Type A ; Chin ; Clostridium botulinum ; Facial Expression ; Foot ; Forehead ; Masks ; Masseter Muscle ; Nasolabial Fold ; Neck ; Neuromuscular Junction ; Paralysis ; Surgery, Plastic*

Botulinum toxin type A is a polypeptide neurotoxin derived from the anaerobic bacterium Clostridium botulinum. Safe and effective Botulinum type A toxin has found clinical application as a treatment for strabismus, blepharospasm, palmar hyperhidrosis. Now this toxin is widely used for improving the facial rhytides cosmetically. Author applicated this toxin not only for facial rejuvenation but also for improving for masseter muscle hypertrophy, facial asymmetries after orthognathic surgery, dysmorphis caused by facial nerve paralysis, TMD and habitual TMJ luxation. Author report clinical results with literature review.]]>
Blepharospasm ; Botulinum Toxins ; Botulinum Toxins, Type A ; Clostridium botulinum ; Facial Asymmetry ; Facial Nerve ; Hyperhidrosis ; Hypertrophy ; Masseter Muscle ; Orthognathic Surgery ; Paralysis ; Rejuvenation ; Strabismus ; Temporomandibular Joint

Designed a pair of primers through modifying N-terminal bases (5bps) of gene after ATG but not changing amino acid, and amplified a smaller mutated gene sequnce (468bp) containing two protective antigenic determinants from pBlue-BoNTaHc, N-terminal codon of mutated gene fragment is changed from low to high frenqence in E. coli. Mutated gene was ligated into pGEM-T vector and sequenced, then, cloned into a expression plasmid pBV220. As a result, cloned gene was expressed in insoluble form by temperature inducing (from 30 degrees C to 42 degrees C) in E. coli. Expression product is 40% of total proteins and is of specific binding activity to antibody in ELISA. The successful modification and high level expression of protective fragment of botulinum neurotoxin serotype A(BoNTaHc468) gene is conducive to further study on antitoxin and vaccine.
Bacterial Vaccines ; immunology ; Botulinum Toxins, Type A ; genetics ; immunology ; Clostridium botulinum type A ; immunology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis

A completely synthetic gene encoding the He domain of Clostridium botulinum neurotoxin serotype A (AHc, 1287 bp, 429 aa, -50 kD) was constructed with oligonucleotides. After expressed in Escherichia coli, soluble product AHc was gained and verified by SDS-PAGE and Western blot analysis. The expressive level of recombinant AHc in E. coli was very high (36%-53% of soluble total proteins) and the purified yield was more than 30 mg/L by one-step purification. Then, the purified AHc was used to vaccinate Balb/c mice, which developed a strong and specific immune response as expected following administration of AHe protein via the subcutaneous route. Results from BoNT/A neutralization assay showed that the serum from mice vaccinated with AHc contained high titer protective antibody. These results showed that the soluble, stable and high-levelly expressive AHc not only could be produced by the prokaryotic expression system built in our lab, but also owned strong immunogenicity to prepare antitoxin for treatment and as sub-unit candidate vaccine for prophylaxis against botulinum toxin serotype A.
Animals ; Antibodies, Bacterial ; blood ; Bacterial Vaccines ; genetics ; immunology ; Botulinum Toxins, Type A ; biosynthesis ; genetics ; immunology ; Botulism ; immunology ; prevention & control ; Clostridium botulinum type A ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; T-Lymphocytes ; immunology ; Vaccines, DNA ; genetics ; immunology