## 4 Results and Discussion
### The Biomolecular Toxicity and Activity of Botulinum Neurooxins (BoNTs)
As mentioned, the action of BoNTs is the effect they have on specific proteins associated with nerve activation, blocking the peripheral release of acetylcholine at presynaptic cholinergic nerve endings through the breakdown of SNAP-25, a protein that is essential for and release of acetylcholine from vesicles found inside nerve endings (Table 2) [27]. Initially the BoNT heavy chain is specifically attached to nerves that use the neurotransmitter acetylcholine. Then the neuron encloses the toxin in a vesicle, which is oxidized as it moves inside the neuron. So, after entering the cytoplasm of the neuron, the light chain zinc-containing protease acts by binding to proteins that play a role in the mechanism of acetylcholine release (SNAP-25, syntaxine, and synaptobevin-VAMP). Thus, the light chain binds to SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins preventing the neuron from releasing the acetylcholine neurotransmitter. The result is that the communication of the nerve cells is blocked, leading to paralysis (Figure 1) [28]. The clinical picture and diagnosis of botulism  
It exists in six main intoxication clinical forms: (a) aimentary, (b) inhaled, (c) wound (incubation period 4-14 days), (d) infant (age 6 \(>\) months), (e) adult intestinal, and (f) iatrogenic. Several improperly made food preparations can provide botulism such as: preserves and semipreserves in oil, water, industrial or domestic brine, the products called RE.P.F.E.D. (Refrigerated Processed Food with Extended Durability), creams and  
Figure 1: Biomolecular mechanism of toxin action: BoNTs (a) have tropism for peripheral motor neurons, (b) cut the SNAP 25 protein, (c) inhibit synaptic vesicle fusion with the presynaptic membrane, (d) inhibit the release of the neurotransmitter acetylcholine at the level of the neuromuscular plate causing flacied paralysis and death from respiratory arrest. BoNTs (e.g., BoNT-A) initially are activated in the digestive system by the action of the trypsin (a serine protease) in the scission region (449-448) and are therefore absorbed by the gut. BoNTs spread in the blood until they reach the neuromuscular and cholinergic junctions, penetrate the synaptic cholinergic endings, and block the release of acetylcholine, causing flacied paralysis. The heavy chain of the toxin is important for its penetration into the axonic extremities, a condition to which the onset of paralysis is linked. Following the binding of the heavy chain to terminal axon proteins, the toxin can enter neurons via endocytosis. The binding of the heavy chain occurs with the protein receptor SV2, whose expression is increased when the synapse is most active. The light chain is able to leave the endocytic vesicles and reach the cytoplasm. The light chain of the toxin possesses protease activity. Toxin A proteolytically degrades SNAP-25 protein, a type of SNARE protein. The SNAP-25 protein is necessary for the release of neurotransmitters from terminal axons (in particular, botulinum toxin degrades SNAREs, preventing the release of neurotransmitters at the synapse level). Credits: Original figure by [PERSON]. \begin{table}
\begin{tabular}{c c} \hline \hline \multicolumn{2}{c}{**BoNT Target Actions**} \\ \hline
**Toxin Isoforms** & **Toxin Targets** \\
**(Neuro–Muscular Junction)** & **(Synapse Proteins)** \\ \hline A, C, E & SNAP-25 \\ B, D, G, F & VAMP (Synaptobrevin) \\ C & Syntaxin \\ \hline \hline \end{tabular}
\end{table} 
Table 2: The action of BoNTs in the Synaptic Fusion Snare Proteins Complex. sauces, some macrobiotic products (seitan or tofu), spices in oil, and flavored oils. Other causes are preserved products, vacuum-packed with equipment available at home, honey, and exposure to water. Symptoms of BoNTs poisoning by inhalation or ingestion usually occur with an incubation period of 12 to 36 h and in exceptional cases up to eight days (maximum time range four hours to eight days) and the onset time is dose-dependent. Thus, a higher dose means a shorter onset of symptoms [29].