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

Analysing agricultural policy reforms under transition in Bulgaria

The elaboration of an appropriate incentive system, including measures of agricultural price and trade policy has important consequences for Bulgarian agriculture. This paper offers a brief picture of the process of reform towards the establishment of a market economy in Bulgaria, as well as of the main developments in its agricultural sector. An analysis is then presented of the impact of alternative agricultural price and trade policy scenarios for the period up to 2002. The results show that the impact of price and trade policies is modest when compared to that of technological change and the increase in incomes. Adoption of the CAP will stimulate production, decrease demand and have strong adverse effects on consumers and taxpayers and wider negative effects on the competitiveness of the food industry. Thus the issue of extending financial support for structural policies rather than granting direct aid requires further investigation. © 2001 Elsevier Science Ltd. All rights reserved

Quality and Shelf-Life Modeling of Frozen Fish at Constant and Variable Temperature Conditions

The objective of this study was the investigation of the effect of variable conditions on quality parameters and the shelf life of fish during frozen storage. Three different fish products were tested, i.e., gilthead sea bream (Sparus aurata) fillets, sea bass (Dicentrarchus labrax) fillets, and yellowfin tuna (Thunnus albacares) slices stored in the range of −5 to −15 °C. The kinetic modeling of different shelf-life indices was conducted. Sensory scoring of frozen fish showed high correlation with color (L-value) and total volatile basic nitrogen (TVBN). The temperature dependence of the rates of quality degradation was expressed via the activation energy values, calculated via the Arrhenius equation, and ranged, for the tested quality indices, between 49 and 84 kJ/mol. The estimated kinetic parameters were validated at dynamic conditions and their applicability in real conditions was established, allowing for their practical application as tools for cold chain management