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The membrane proximal region (MPR, residues 649–683) and transmembrane domain (TMD, residues 684–705) of the gp41 subunit of HIV-1’s envelope protein are highly conserved and are important in viral mucosal transmission, virus attachment and membrane fusion with target cells. Several structures of the trimeric membrane proximal external region (residues 662–683) of MPR have been reported at the atomic level; however, the atomic structure of the TMD still remains unknown. To elucidate the structure of both MPR and TMD, we expressed the region spanning both domains, MPR-TM (residues 649–705), in Escherichia coli as a fusion protein with maltose binding protein (MBP). MPR-TM was initially fused to the C-terminus of MBP via a 42 aa-long linker containing a TEV protease recognition site (MBP-linker-MPR-TM).
Biophysical characterization indicated that the purified MBP-linker-MPR-TM protein was a monodisperse and stable candidate for crystallization. However, crystals of the MBP-linker-MPR-TM protein could not be obtained in extensive crystallization screens. It is possible that the 42 residue-long linker between MBP and MPR-TM was interfering with crystal formation. To test this hypothesis, the 42 residue-long linker was replaced with three alanine residues. The fusion protein, MBP-AAA-MPR-TM, was similarly purified and characterized. Significantly, both the MBP-linker-MPR-TM and MBP-AAA-MPR-TM proteins strongly interacted with broadly neutralizing monoclonal antibodies 2F5 and 4E10. With epitopes accessible to the broadly neutralizing antibodies, these MBP/MPR-TM recombinant proteins may be in immunologically relevant conformations that mimic a pre-hairpin intermediate of gp41.
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The positive impacts of yoga on stress, pain, and chronic disease has recently led to the integration of yoga as part of physical therapy (PT) treatment. Due to the lack of training for PTs related to yoga, there is currently a need to provide knowledge and education about how to safely and easily implement therapeutic yoga (TY) as a complementary treatment approach.
Objective:
The purpose of this study was to assess the readiness of PTs (those who do not currently prescribe TY to patients) to integrate TY into treatment, and secondly, the feasibility (i.e., acceptability, demand, and practicality) of a 5-week online TY training to improve the readiness of PT’s to utilize TY in their practice.
Methods:
Licensed Physical Therapist’s (n=103) were recruited nationally through social media and email. Eligible and consented participants were asked to register in a 5-week online TY training course, Readiness for Integrating Yoga Therapeutics into Rehabilitation for PTs (intervention). PTs perceptions of TY and the role of safety and confidence in prescribing TY to patients were measured at baseline and post-intervention using a customized survey. Feasibility outcomes were measured after completion of the 5-week online training course with a survey. Feasibility was measured with acceptability, demand, and practicality. Our benchmarks included: (1) at least 70% of PTs would find the course acceptable, (2) at least 60% would finish the course (i.e., demand) and (3) there would be significant improvements in PTs perceptions of TY.
Results:
A total of 95 licensed PTs registered in the 5-week online TY training course, with 60 PTs (63%) completing the intervention and surveys. Of the PTs who completed the 5-week online training course, most PTs felt they were not ready (n=19/60, 31.7%) or somewhat ready (n=25/60, 41.7%) to integrate TY prior to taking the online training. Over half of PTs thought the online training was acceptable (n= 50/60, 83.3%) and finished the course (n=60/95, 63%). There were significant improvements in personal readiness to prescribe TY, safety prescribing TY, confidence to prescribe TY, current understanding/knowledge of TY and feeling adequately trained and educated to use some form of TY techniques with patients.
Conclusion:
Findings suggest a 5-week online TY training course is feasible in improving PTs readiness to prescribe TY, safety prescribing TY, confidence to prescribe TY, current understanding/knowledge of TY and feeling adequately trained and educated to use some form of TY techniques with patients. Future studies are proposed to test the effectiveness of TY training and education opportunities with PTs to further advance the adoption of TY into PT practice.