Methods: This is a community-based study conducted among adults between 2010 and 2011 among a rural population in Raub, Pahang, Malaysia. Blood pressure was measured after 5 min of rest. Measurement was done twice and the average was recorded. Cardiovascular risk perception score (CvRPS) was derived using the Modified Risk and Health Behavior Questionnaire. Higher CvRPS indicates the respondent perceives a poorer prognostic outlook.
Results: A total of 383 respondents who have hypertension participated in this study. The mean age of respondents was 62±10.6 years; men 63.1±9.6 years, women 61.2±11.1 years (p>0.05). Among hypertensives, those who were not on medication had significantly lower CvRPS compared with those who were on medications (115.9±22.1vs 120.9±23.5, p=0.036); those who were not aware of their hypertensive status had significantly lower CvRPS compared with respondents who were aware about their hypertension (116.7±22.5vs 121.7±21.3, p=0.029) and those with uncontrolled hypertension had significantly lower CvRPS compared with those whose blood pressure was controlled (118.2±22.2vs 128.8±25.8, p=0.009).
Conclusions: Our study shows that respondents who were not on medications, unaware of their hypertension status and those who had uncontrolled hypertension tended to underestimate (lower CvRPS) their risk for CVD. Improving their CvPRS through a concerted health education may lead to better therapeutic behaviour and outcomes.
RESULTS: A novel approach of utilizing an in-trans protein surface display system of Lactobacillus plantarum carrying and displaying combination of Mycobacterium tuberculosis subunit epitope antigens (Ag85B, CFP-10, ESAT-6, Rv0475 and Rv2031c) fused with LysM anchor motif designated as ACERL was constructed, cloned and expressed in Esherichia coli Rossetta expression host. Subsequently the binding capability of ACERL to the cell wall of L. plantarum was examined via the immunofluorescence microscopy and whole cell ELISA where successful attachment and consistent stability of cell wall binding up to 4 days was determined. The immunization of the developed vaccine of L. plantarum surface displaying ACERL (Lp ACERL) via the oral route was studied in mice for its immunogenicity effects. Lp ACERL immunization was able to invoke significant immune responses that favor the Th1 type cytokine response of IFN-γ, IL-12 and IL-2 as indicated by the outcome from the cytokine profiling of spleen, lung, gastrointestinal tract (GIT), and the re-stimulation of the splenocytes from the immunized mice. Co-administration of an adjuvant consisting of Lactococcus lactis secreting mouse IL-12 (LcIL-12) with Lp ACERL was also investigated. It was shown that the addition of LcIL-12 was able to further generate significant Th1 type cytokines immune responses, similar or better than that of Lp ACERL alone which can be observed from the cytokine profiling of the immunized mice's spleen, lung and GIT.
CONCLUSIONS: This study represents a proof of concept in the development of L. plantarum as a carrier for a non-genetically modified organism (GMO) tuberculosis vaccine, which may be the strategy in the future for tuberculosis vaccine development.
RESULTS: Expression of TB antigen-LysM fusion genes was conducted in Escherichia coli, but this resulted in IBs deposition in contrast to the expression of TB antigens only. This suggested that LysM fusion significantly altered solubility of the TB antigens produced in E. coli. The non-denaturing NLS technique was used and optimized to successfully solubilize and purify ~ 55% of the recombinant cell wall-anchoring TB antigen from the IBs. Functionality of the recovered protein was analyzed via immunofluorescence microscopy and whole cell ELISA which showed successful and stable cell wall binding to L. plantarum (up to 5 days).
CONCLUSION: The presented NLS purification strategy enables an efficient and rapid method for obtaining higher yields of soluble cell wall-anchoring Mycobacterium tuberculosis antigens-LysM fusion proteins from IBs in E. coli.
RESULTS: Xylanase was successfully expressed in Lactococcus lactis. Recombinant xylanase fused to either signal peptide Usp45 or Spk1 showed halo zone on Remazol Brilliant Blue-Xylan plates. This indicated that the xylanase was successfully secreted from the cell. The culture supernatants of strains secreting the xylanase with help of the Spk1 and Usp45 signal peptides contained 49.7 U/ml and 34.4 U/ml of xylanase activity, respectively.
CONCLUSION: Although Usp45 is the most commonly used signal peptide when secreting heterologous proteins in Lactococcus lactis, this study shows that Spk1 isolated from Pediococcus pentosaceus was superior to Usp45 in regard to xylanase protein secretion.