Thermal Stability of the Human Immunodeficiency Virus Type 1 (HIV-1) Receptors, CD4 and CXCR4, Reconstituted in Proteoliposomes

BACKGROUND: The entry of human immunodeficiency virus (HIV-1) into host cells involves the interaction of the viral exterior envelope glycoprotein, gp120, and receptors on the target cell. The HIV-1 receptors are CD4 and one of two chemokine receptors, CCR5 or CXCR4. METHODOLOGY/PRINCIPAL FINDINGS: We created proteoliposomes that contain CD4, the primary HIV-1 receptor, and one of the coreceptors, CXCR4. Antibodies against CD4 and CXCR4 specifically bound the proteoliposomes. CXCL12, the natural ligand for CXCR4, and the small-molecule CXCR4 antagonist, AMD3100, bound the proteoliposomes with affinities close to those associated with the binding of these molecules to cells expressing CXCR4 and CD4. The HIV-1 gp120 exterior envelope glycoprotein bound tightly to proteoliposomes expressing only CD4 and, in the presence of soluble CD4, bound weakly to proteoliposomes expressing only CXCR4. The thermal stability of CD4 and CXCR4 inserted into liposomes was examined. Thermal denaturation of CXCR4 followed second-order kinetics, with an activation energy (E(a)) of 269 kJ/mol (64.3 kcal/mol) and an inactivation temperature (T(i)) of 56°C. Thermal inactivation of CD4 exhibited a reaction order of 1.3, an E(a) of 278 kJ/mol (66.5 kcal/mol), and a T(i) of 52.2°C. The second-order denaturation kinetics of CXCR4 is unusual among G protein-coupled receptors, and may result from dimeric interactions between CXCR4 molecules. CONCLUSIONS/SIGNIFICANCE: Our studies with proteoliposomes containing the native HIV-1 receptors allowed an examination of the binding of biologically important ligands and revealed the higher-order denaturation kinetics of these receptors. CD4/CXCR4-proteoliposomes may be useful for the study of virus-target cell interactions and for the identification of inhibitors

Omics-based molecular techniques in oral pathology centred cancer: Prospect and challenges in Africa

: The completion of the human genome project and the accomplished milestones in the human proteome project; as well as the progress made so far in computational bioinformatics and “big data” processing have contributed immensely to individualized/personalized medicine in the developed world.At the dawn of precision medicine, various omics-based therapies and bioengineering can now be applied accurately for the diagnosis, prognosis, treatment, and risk stratifcation of cancer in a manner that was hitherto not thought possible. The widespread introduction of genomics and other omics-based approaches into the postgraduate training curriculum of diverse medical and dental specialties, including pathology has improved the profciency of practitioners in the use of novel molecular signatures in patient management. In addition, intricate details about disease disparity among diferent human populations are beginning to emerge. This would facilitate the use of tailor-made novel theranostic methods based on emerging molecular evidences

Environmental measures of physical activity supports: Perception versus reality

Background Perceptions of the environment and physical activity have been associated using survey methods, yet little is known about the validity of environmental surveys. In this study, perceptions of the environment at neighborhood and community levels were assessed (1) to determine validity by comparing respondent perceptions to objective measures and (2) to determine test–retest reliability of the survey. Methods A telephone survey was administered to a stratified sample of Sumter County, South Carolina adults. Respondents’ home addresses were mapped using a geographic information system (GIS) (n =1112). As an indicator of validity, kappa statistics were used to measure agreement between perceptions and objective measures identified at neighborhood and community levels using GIS. A second survey in an independent sample (n=408) assessed test–retest reliability. Results When assessing perceptions of environmental and physical activity in a defined geographic area, validity and reliability for neighborhood survey items were κ=−0.02 to 0.37 and RHO=0.42 to 0.74, and for community survey items were κ=−0.07 to 0.25 and RHO=0.28 to 0.56. Conclusions Although causality between perception of access and safety and actual physical activity level cannot be assumed, those meeting national physical activity guidelines or reporting some physical activity demonstrated greatest agreement with access to recreation facilities, while those not meeting the guidelines demonstrated greater agreement with safety of recreation facilities. Factors such as distance and behavior may explain differences in perceptions at neighborhood and community levels. Using local environments with short distances in survey methods improves validity and reliability of results