Expression, Purification and Characterization of GMZ2'.10C, a Complex Disulphide-Bonded Fusion Protein Vaccine Candidate against the Asexual and Sexual Life-Stages of the Malaria-Causing Plasmodium falciparum Parasite

PURPOSE: Production and characterization of a chimeric fusion protein (GMZ2'.10C) which combines epitopes of key malaria parasite antigens: glutamate-rich protein (GLURP), merozoite surface protein 3 (MSP3), and the highly disulphide bonded Pfs48/45 (10C). GMZ2'.10C is a potential candidate for a multi-stage malaria vaccine that targets both transmission and asexual life-cycle stages of the parasite. METHODS: GMZ2'.10C was produced in Lactococcus lactis and purified using either an immunoaffinity purification (IP) or a conventional purification (CP) method. Protein purity and stability was analysed by RP-HPLC, SEC-HPLC, 2-site ELISA, gel-electrophoresis and Western blotting. Structural characterization (mass analysis, peptide mapping and cysteine connectivity mapping) was performed by LC-MS/MS. RESULTS: CP-GMZ2'.10C resulted in similar purity, yield, structure and stability as compared to IP-GMZ2'.10C. CP-GMZ2'.10C and IP-GMZ2'.10C both elicited a high titer of transmission blocking (TB) antibodies in rodents. The intricate disulphide-bond connectivity of C-terminus Pfs48/45 was analysed by tandem mass spectrometry and was established for GMZ2'.10C and two reference fusion proteins encompassing similar parts of Pfs48/45. CONCLUSION: GMZ2'.10C, combining GMZ2' and correctly-folded Pfs48/45 can be produced by the Lactoccus lactis P170 based expression system in purity and quality for pharmaceutical development and elicit high level of TB antibodies. The cysteine connectivity for the 10C region of Pfs48/45 was revealed experimentally, providing an important guideline for employing the Pfs48/45 antigen in vaccine design

Aspects of parenting and family functioning associated with obesity in adolescent refugees and migrants from African backgrounds living in Australia

Objective: To investigate the relationship between parenting style and family functioning, and BMI, among adolescent migrants and refugees from African countries.Method: A total of 104 parents and their adolescent offspring completed questionnaires assessing the variables of interest, and anthropometric data were collected from them.Results: Parents reported higher maternal involvement, lower levels of paternal involvement, higher levels of positive parenting, greater use of ‘other’ (non-corporal) discipline styles, greater satisfaction and better communication than did their offspring. Parents also reported greater family cohesion and flexibility than adolescents, and saw their families as more enmeshed and more rigid. Parenting style and family functioning were not strong predictors of BMI according to either report. For adolescents, inconsistent discipline and lack of parental supervision accounted for significant variance in BMI.Conclusions: Inconsistent discipline and lack of parental supervision may be related to adolescent BMI. Further studies with larger samples are required to confirm these relationships with the view to informing obesity prevention programs for this target population.Implications: Prevention or intervention programs aimed at reducing the prevalence of overweight and obesity need to consider parenting style in their design