Search

Technical and methodological advances in single-particle cryo-electron microscopy (cryo-EM) have expanded the technique into a resolution regime that was previously only attainable by X-ray crystallography. Although single-particle cryo-EM has proven to be a useful technique for determining the structures of biomedically relevant molecules at near-atomic resolution, nearly 98% of the structures resolved to better than 4 Å resolution have been determined using 300 keV transmission electron microscopes (TEMs). We demonstrate that it is possible to obtain cryo-EM reconstructions of macromolecular complexes at a range of sizes to better than 3 Å resolution using a 200 keV TEM. These structures are of sufficient quality to unambiguously assign amino acid rotameric conformations and identify ordered water molecules, features previously thought only to be resolvable using TEMs operating at 300 keV.

The accidental leakage of long-distance pressurized oil pipelines is a major area of risk, capable of causing extensive damage to human health and environment. However, the complexity of the leaking process, with its complex boundary conditions, leads to difficulty in calculating the leakage volume. In this study, the leaking process is divided into 4 stages based on the strength of transient pressure. 3 models are established to calculate the leaking flowrate and volume. First, a negative pressure wave propagation attenuation model is applied to calculate the sizes of orifices. Second, a transient oil leaking model, consisting of continuity, momentum conservation, energy conservation and orifice flow equations, is built to calculate the leakage volume. Third, a steady-state oil leaking model is employed to calculate the leakage after valves and pumps shut down. Moreover, sensitive factors that affect the leak coefficient of orifices and volume are analyzed respectively to determine the most influential one. To validate the numerical simulation, two types of leakage test with different sizes of leakage holes were conducted from Sinopec product pipelines. More validations were carried out by applying commercial software to supplement the experimental insufficiency. Thus, the leaking process under different leaking conditions are described and analyzed.

SUMMARY Understanding how broadly neutralizing antibodies (bnAbs) to HIV envelope (Env) develop during natural infection can help guide the rational design of an HIV vaccine. Here, we described a bnAb lineage targeting the Env V2 apex and the Ab-Env co-evolution that led to development of neutralization breadth. The lineage Abs bore an anionic heavy chain complementarity-determining region 3 (CDRH3) of 25 amino acids, among the shortest known for this class of Abs, and achieved breadth with only 10% nucleotide somatic hypermutation and no insertions or deletions. The data suggested a role for Env glycoform heterogeneity in the activation of the lineage germ-line B cell. Finally, we showed that localized diversity at key V2 epitope residues drove bnAb maturation toward breadth, mirroring the Env evolution pattern described for another donor who developed V2-apex targeting bnAbs. Overall, these findings suggest potential strategies for vaccine approaches based on germline-targeting and serial immunogen design., In Brief Understanding the molecular basis of HIV Env-specific broadly neutralizing antibodies (bnAbs) development is key for vaccine design. Landais et al. find that glycan heterogeneity played a role in the activation of V2 apex PCT64 bnAbs precursor and that viral evolution was similar to CAP256, another donor with V2 apex bnAbs.