RESUMEN
We study how individual decisions are affected by those of other members of the society. We use the vaccine against COVID-19 as a case study and empirically estimate the magnitude of three key forces: Herding, Social Norms, and Free-riding. We find that Free-riding is dominated by the other two forces, and that Social Norms are a key driver of behavior. There is, however, substantial heterogeneity and systematic differences between people by demographics and their political preferences.
RESUMEN
The pursuit of fully autotrophic nitrogen removal via the anaerobic ammonium oxidation (anammox) pathway has led to an increased interest in carbon removal technologies, particularly the A-stage of the adsorption/bio-oxidation (A/B) process. The high-rate operation of the A-stage and lack of automatic process control often results in wide variations of chemical oxygen demand (COD) removal that can ultimately impact nitrogen removal in the downstream B-stage process. This study evaluated the use dissolved oxygen (DO) and mixed liquor suspended solids (MLSS) based automatic control strategies through the use of in situ on-line sensors in the A-stage of an A/B pilot study. The objective of using these control strategies was to reduce the variability of COD removal by the A-stage and thus the variability of the effluent C/N. The use of cascade DO control in the A-stage did not impact COD removal at the conditions tested in this study, likely because the bulk DO concentration (>0.5 mg/L) was maintained above the half saturation coefficient of heterotrophic organisms for DO. MLSS-based solids retention time (SRT) control, where MLSS was used as a surrogate for SRT, did not significantly reduce the effluent C/N variability but it was able to reduce COD removal variation in the A-stage by 90%.
Asunto(s)
Análisis de la Demanda Biológica de Oxígeno , Carbono/metabolismo , Nitrógeno/metabolismo , Oxígeno/metabolismo , Eliminación de Residuos Líquidos/instrumentación , Reactores Biológicos , Oxidación-Reducción , Proyectos Piloto , Eliminación de Residuos Líquidos/métodosRESUMEN
BACKGROUND: HIV-1 entry into host cells is mediated by interactions between the virus envelope glycoprotein (gp120/gp41) and host-cell receptors. N-glycans represent approximately 50% of the molecular mass of gp120 and serve as potential antigenic determinants and/or as a shield against immune recognition. We previously reported that N-glycosylation of recombinant gp120 varied, depending on the producer cells, and the glycosylation variability affected gp120 recognition by serum antibodies from persons infected with HIV-1 subtype B. However, the impact of gp120 differential glycosylation on recognition by broadly neutralizing monoclonal antibodies or by polyclonal antibodies of individuals infected with other HIV-1 subtypes is unknown. METHODS: Recombinant multimerizing gp120 antigens were expressed in different cells, HEK 293T, T-cell, rhabdomyosarcoma, hepatocellular carcinoma, and Chinese hamster ovary cell lines. Binding of broadly neutralizing monoclonal antibodies and polyclonal antibodies from sera of subtype A/C HIV-1-infected subjects with individual gp120 glycoforms was assessed by ELISA. In addition, immunodetection was performed using Western and dot blot assays. Recombinant gp120 glycoforms were tested for inhibition of infection of reporter cells by SF162 and YU.2 Env-pseudotyped R5 viruses. RESULTS: We demonstrated, using ELISA, that gp120 glycans sterically adjacent to the V3 loop only moderately contribute to differential recognition of a short apex motif GPGRA and GPGR by monoclonal antibodies F425 B4e8 and 447-52D, respectively. The binding of antibodies recognizing longer peptide motifs overlapping with GPGR epitope (268 D4, 257 D4, 19b) was significantly altered. Recognition of gp120 glycoforms by monoclonal antibodies specific for other than V3-loop epitopes was significantly affected by cell types used for gp120 expression. These epitopes included CD4-binding site (VRC03, VRC01, b12), discontinuous epitope involving V1/V2 loop with the associated glycans (PG9, PG16), and an epitope including V3-base-, N332 oligomannose-, and surrounding glycans-containing epitope (PGT 121). Moreover, the different gp120 glycoforms variably inhibited HIV-1 infection of reporter cells. CONCLUSION: Our data support the hypothesis that the glycosylation machinery of different cells shapes gp120 glycosylation and, consequently, impacts envelope recognition by specific antibodies as well as the interaction of HIV-1 gp120 with cellular receptors. These findings underscore the importance of selection of appropriately glycosylated HIV-1 envelope as a vaccine antigen.
RESUMEN
Human immunodeficiency virus type 1 (HIV-1) entry is mediated by the interaction between a variably glycosylated envelope glycoprotein (gp120) and host-cell receptors. Approximately half of the molecular mass of gp120 is contributed by N-glycans, which serve as potential epitopes and may shield gp120 from immune recognition. The role of gp120 glycans in the host immune response to HIV-1 has not been comprehensively studied at the molecular level. We developed a new approach to characterize cell-specific gp120 glycosylation, the regulation of glycosylation, and the effect of variable glycosylation on antibody reactivity. A model oligomeric gp120 was expressed in different cell types, including cell lines that represent host-infected cells or cells used to produce gp120 for vaccination purposes. N-Glycosylation of gp120 varied, depending on the cell type used for its expression and the metabolic manipulation during expression. The resultant glycosylation included changes in the ratio of high-mannose to complex N-glycans, terminal decoration, and branching. Differential glycosylation of gp120 affected envelope recognition by polyclonal antibodies from the sera of HIV-1-infected subjects. These results indicate that gp120 glycans contribute to antibody reactivity and should be considered in HIV-1 vaccine design.
Asunto(s)
Anticuerpos/inmunología , Proteína gp120 de Envoltorio del VIH/inmunología , Proteína gp120 de Envoltorio del VIH/metabolismo , Síndrome de Inmunodeficiencia Adquirida/inmunología , Especificidad de Anticuerpos , Línea Celular , Línea Celular Tumoral , ADN Complementario/genética , Ensayo de Inmunoadsorción Enzimática/métodos , Glicósido Hidrolasas/metabolismo , Glicosilación , Proteína gp120 de Envoltorio del VIH/genética , Proteína gp120 de Envoltorio del VIH/aislamiento & purificación , Seropositividad para VIH/inmunología , Seropositividad para VIH/metabolismo , VIH-1/inmunología , VIH-1/metabolismo , Células Hep G2/metabolismo , Humanos , Células Jurkat/metabolismo , Manosa/metabolismo , Lectina de Unión a Manosa/genética , Espectrometría de Masas , Oligosacáridos/química , Oligosacáridos/aislamiento & purificación , Plásmidos , Polisacáridos/química , Polisacáridos/aislamiento & purificaciónRESUMEN
The specificities of 65 compounds reported to be relatively specific inhibitors of protein kinases have been profiled against a panel of 70-80 protein kinases. On the basis of this information, the effects of compounds that we have studied in cells and other data in the literature, we recommend the use of the following small-molecule inhibitors: SB 203580/SB202190 and BIRB 0796 to be used in parallel to assess the physiological roles of p38 MAPK (mitogen-activated protein kinase) isoforms, PI-103 and wortmannin to be used in parallel to inhibit phosphatidylinositol (phosphoinositide) 3-kinases, PP1 or PP2 to be used in parallel with Src-I1 (Src inhibitor-1) to inhibit Src family members; PD 184352 or PD 0325901 to inhibit MKK1 (MAPK kinase-1) or MKK1 plus MKK5, Akt-I-1/2 to inhibit the activation of PKB (protein kinase B/Akt), rapamycin to inhibit TORC1 [mTOR (mammalian target of rapamycin)-raptor (regulatory associated protein of mTOR) complex], CT 99021 to inhibit GSK3 (glycogen synthase kinase 3), BI-D1870 and SL0101 or FMK (fluoromethylketone) to be used in parallel to inhibit RSK (ribosomal S6 kinase), D4476 to inhibit CK1 (casein kinase 1), VX680 to inhibit Aurora kinases, and roscovitine as a pan-CDK (cyclin-dependent kinase) inhibitor. We have also identified harmine as a potent and specific inhibitor of DYRK1A (dual-specificity tyrosine-phosphorylated and -regulated kinase 1A) in vitro. The results have further emphasized the need for considerable caution in using small-molecule inhibitors of protein kinases to assess the physiological roles of these enzymes. Despite being used widely, many of the compounds that we analysed were too non-specific for useful conclusions to be made, other than to exclude the involvement of particular protein kinases in cellular processes.