Botulinum complex

The fiberoptic biosensor used in the detection of type E botulinum complex consists of five main components the sensor (7 subcomponents), the detector (3 subcomponents), the lock-in amplifier, the laser power supply and a lap-top computer. A schematic representation of all of the biosensor s components and subcomponents with illustration of the laser light path and the returning fluorescent signal is shown in Figure 1. 

The detection of type E C botulinum complex using the fiberoptic biosensor was accomplished using a two-step sandwich immunoassay. The assay involved (1) the incubation of antibody bound fibers in a solution of type E complex followed by (2) signal collection upon incubation of the resulting complex bound fibers in a fluorescently labeled antibody solution. The procedures and fundamentals involved are discussed below in detail. 

Figure 5. Gel electrophoresis of type E Clostridium botulinum complex. Lane 1 = molecular weight standards. Lane 2= type E complex 1st band = 150 kDa toxin band, 2nd band = 120 kDa NBP band.

Bacteria, viruses, and rickettsiae have similar symptom progressions in that exposure is followed by a period of reproductive growth (often nonsympto-matic) in the body. As their numbers increase, they often eventually overcome the immune system. Many produce toxins that interfere with bodily functions. Purified toxins such as botulinum toxin (produced by the Clostridium botulinum bacteria) act in a similar manner to chemical agents since, as complex chemical compounds, they do not reproduce but immediately interfere with bodily functions. However, most toxins are not absorbed through the skin, as... 

Bullens, R. W., O Hanlon, G. M., Wagner, E. etal. Complex gangliosides at the neuromuscular junction are membrane receptors for autoantibodies and botulinum neurotoxin but redundant for normal synaptic function. /. Neurosci. 22 6876-6884, 2002. 

Botulin toxin may also act as an inhalation poison. This property has been reveled for both pure and progenitor (a complex with auxiliary proteins) toxins (Park and Simpson, 2003). C. botulinum and its toxin were included onto the list of high-priority agents - in the highest A category of agents that may potentially be used in bioterrorism (Khan et al., 2000 Sobel et al., 2002). 

The outgrowth of C. botulinum requires a suitable medium, temperature, atmosphere, pH, Eh potential, and water activity. Toxin is usually only produced in optimal or close-to-optimal conditions. Nutrient demands of C. botulinum are complex, and include amino acids, B vitamins, and minerals. In broth, non-proteolytic strains of type B and F grow and produce toxin at 4°C, but in crab meat the outgrowth and toxin production occurs solely at 26°C (Alberto et al., 2003). 

East, A.K., Stacey, I.M. and Collins, M.D., Cloning and sequencing of hemagglutinin component of the botulinum neurotoxin complex encoded by Clostridium botulinum types A and B, Syst. Appl. Microbiol., 17, 306-312, 1994. 

Powder for injection (vacuum-dried) 100 units of vacuum-dried Clostridium botulinum toxin type A neurotoxin complex- (Rx)... 

Other Botulinum toxin A, neutrotoxin complex Botox Vacuum dried 100 U vial IM NA... 

Reddy, D., Lancaster, J. R., Jr., and Comforth, D. P. (1983). Nitrite inhibition of Clostridium botulinum Electron spin resonance detection of iron-nitric oxide complexes. Science 221, 769-770. 

The complex of SNAREs and SNAP25 is the target of the powerful Clostridium, botulinum, toxin, a protease that cleaves specific bonds in these proteins, preventing neurotransmission and causing the death of the organism. Because of its very high specificity for these proteins, purified botulinum toxin has served as a powerful tool for dissecting the mechanism of neurotransmitter release in vivo and in vitro. 

More important than the demonstration (107) of nitrosylation of synthetic model clusters is the demonstration (108) of nitrosylation by nitrite of the [4Fe-4S] center in vegetative cells of Clostridium botulinum. Treatment of a reduced culture of C. botulinum with nitrite in the presence of ascorbate resulted in loss of the ESR signal at g = 1.94, characteristic of the reduced iron-sulfur protein, and its replacement by a signal at g — 2.035, characteristic of [Fe(NO)2X2] complexes, most likely of the [Fe(NO)2(SR)2] type. 

Botulinum toxin A complexed with haemagglutinin is currently employed medicinally to counter involuntary facial muscle spasms, e.g. around the eye. Very small (nanogram) amounts are injected locally and result in the destruction of the acetylcholine release mechanism at the neuromuscular junction. Since new nerve junctions will gradually be formed over two months or so, the result is not permanent, and the treatment will need to be repeated. It has also been found useful in easing muscle spasticity in children with cerebral palsy. 

Chen F, Kuziemko GM, Amersdorfer P, Wong C, Marks JD et al. (1997) Antibody mapping to domains of botulinum neurotoxin serotype A in the complexed and uncomplexed forms. Infect Immun 65 1626-30... 

Chen F, Kuziemko GM, Stevens RC (1998a) Biophysical characterization of the stability of the 150-kilodalton botulinum toxin, the nontoxic component, and the 900-kilodalton botulinum toxin complex species. Infect Immun 66 2420-5... 

Reilich P, Fheodoroff K, Kern U, Mense S, Seddigh S et al. (2004) Consensus statement botulinum toxin in myofascial [corrected] pain. J Neurol 251 Suppl 1 136-8 Rickman C, Davletov B (2005) Arachidonic acid allows SNARE complex formation in the presence of Muncl8. Chem Biol 12 545-53... 

The accessory proteins of the progenitor toxin serve to enhance the stability of the toxin to ensure uptake from the gut. The NTNH component of the multimeric type A toxin complex is encoded by a single gene upstream of the neurotoxin locus while three different HA proteins have been characterized in association with BoNTs. All C. botulinum serotypes have been shown to produce neurotoxin complexes with the NTNH and HA proteins. 

---