Expression and Purification of Botulinum Neurotoxin Type ‘A’ Toxin Domain in E. coli and its Application in Detection

Botulism is a neuroparalytic disease caused by botulinum toxins type A-G. It has been listed as category ‘A’ biowarfare agent by CDC due to its extreme toxicity.. In the present study, we developed a simple method for the pro-duction and purification of recombinant toxin domain of BoNT type ‘A’ from Escherichia coli and established its application in BoNT detection. The BoNT/A LC gene was cloned in pET28b+ vector and expressed in E.coli. The recom-binant BoNT/A LC protein expression was achieved at 25°C with the induc-tion of 0.5mM IPTG at 16 h. The recombinant protein was purified under native conditions with more than 98% purity using simple affinity chroma-tography and confirmed by Mass spectrometry and Western blot analysis. Generated polyclonal antibodies and the toxin domain elicited a strong im-mune response in mice and rabbit. Optimized the Sandwich ELISA system resulted with the sensitivity of 7.5 ng/ml for the detection of BoNT/A. The limit of detection (LOD) in different food matrices were studied through spik-ing studies using rBoNT/A LC protein as antigen and achieved a detection limit ranging from7.5 ng/ml to 250 ng/ml. The developed method has the potential for the industrial production towards its application in detection, development of candidate vaccine molecule and production of neutralizing antibodies. Current study explored for detection of BoNT/A.

Botulinum Toxins and Radio Frequency for Facial Rejuvenation

Botulinum toxins type A injections are likely exceeding 10 million yearly worldwide. While it is easy to “push the plunger” to achieve some result, precise, efficacious, and safe application for functional and aesthetic enhancement is deceptively more challenging. Accurate physical assessment is critical. Advanced techniques are surprisingly more difficult, and the pharmacology of toxins must be understood. Radio frequency nerve ablation is a newer approach that requires even more knowledge, precision, and preprocedure preparation while offering longer or permanent duration and the ability to achieve real-time results. In addition, it can be combined easily with surgical procedures.

Six-Month Comparative Analysis Monitoring the Progression of the Largest Diameter of the Sweating Inhibition Halo of Different Botulinum Toxins Type-A

Background Excessive sweating is a clinical condition that can be improved with type-A botulinum toxin (BTX-A). Objectives To evaluate and compare the largest diameter of sweating inhibition halo (SIH) of 5 different commercially available BTX-A, in five different doses, in a 6-month-long clinical evaluation. Methods Twenty-five adult female volunteers were injected in the dorsal trunk area with both 100 units (100UI) and 500 units (500UI) BTX-A products, reconstituted in a ratio of 1:2.5 IU, respectively. Products were applied in five different concentrations (1:2.5U, 2:5U, 3:7.5U, 4:10U, and 5:12.5U). After 30, 60, 90, 120, 150, and 180 days, a starch-iodine test was performed to obtain the largest diameter of each SIH. Results The higher the number of units used, the larger the SIH p < 0.05 for all BTX-A. However, Botox®, Botulift®, Dysport®, and Prosigne® have pretty likewise SIH along the study, with some few differences for some doses and months between one and another. However, Xeomin® is the BTX-A with the smallest SIH, in comparison with all others, in any dose and period. Conclusions Differences were observed among all brands of BTX-As, based on dose and time after injection. Xeomin® provides the smallest SIH in comparison with others BTX-A. Level of Evidence: 2

Conservative treatment of pronational contracture of the forearm in children with cerebral palsy

Aim. We aimed to evaluate the effectiveness of conservative treatment for pronational contracture of the forearm, depending on the contracture severity. Materials and methods. This study was based on the results of the examination and treatment of children experiencing cerebral palsy with upper limb involvement. The main criterion for patient selection was the presence of pronational contracture of the forearm, both isolated and in combination with other contractures in the upper limb joints. Three patient groups were formed based on the pronounced pronation contracture of the forearm. Results and conclusions. Previous research has established that with the increase in the degree of severity of pronation contracture, the effectiveness of conservative treatment decreases in general. Conservative treatment of patients with a deficiency of active supination of the forearm of > 90° is ineffective. The use of botulinum toxins type A and RFD in groups II and III is ineffective. Conservative treatment with botulinum toxins type A achieved good results only in patients of group I. The RFD method had a longer lasting; however, there were significantly more complications. The use of botulinum toxins type A (m. pronator teres) among patients with pronational contracture of the forearm with the possibility of active supination of the forearm > 90° significantly improved the result of basic conservative treatment.