Evaluation of bidirectional causal association between cardiovascular diseases and pneumonia: A Mendelian randomization study.

AIMS: Observational evidence suggests a bidirectional relationship between cardiovascular diseases (CVDs) and pneumonia. However, the causality between CVDs and pneumonia remains undetermined. Thus, we aimed to investigate the bidirectional causality between CVDs and pneumonia using Mendelian randomization (MR) analysis. METHODS: Global genetic correlation analysis and bidirectional two-sample MR analysis were performed to infer the genetic correlation and causality between CVDs and pneumonia by using genome-wide association study (GWAS) summary data from GWAS meta-analysis study, FinnGen or UK Biobank consortium. Post-hoc power calculation was conducted to assess the power for detecting the causality. RESULTS: The linkage disequilibrium score regression analysis suggested a positive significant genetic correlation between CVDs and pneumonia. In the MR analysis, only genetically predicted ischemic stroke was causally associated with any pneumonia (odds ratio [OR]: 1.119, 95% confidence interval [CI]: 1.031-1.393), bacterial pneumonia (OR: 1.251, 95% CI: 1.032-1.516), and pneumococcal pneumonia (OR: 1.308, 95% CI: 1.093-1.565), but the causality was attenuated to non-significance after adjusting for deep venous thrombosis. However, the causal effects of pneumonia on CVDs were not detected. Post-hoc power calculations supported strong power (more than 80%) to detect the causality. CONCLUSIONS: Ischemic stroke is causally associated with an increased risk of pneumonia, but there is no evidence for the causal effect of pneumonia on CVDs. Our findings have important implications as they provide further support for the thrombosis risk screening as a strategy to reduce the incidence of pneumonia in patients with ischemic stroke.

This Mendelian randomization analysis aimed to investigate the bidirectional causality between cardiovascular diseases and pneumonia. Our findings support the causal association of ischemic stroke on pneumonia, but indicate no evidence for the causal effects of pneumonia on cardiovascular diseases. The causal association of ischemic stroke on pneumonia was revealed to rely on deep venous thrombosis, which provided further support for the thrombosis risk screening as a strategy to reduce the incidence of pneumonia in patients with ischemic stroke.

Cotyledon peeling method for passion fruit protoplasts: a versatile cell system for transient gene expression in passion fruit (Passiflora edulis).

Passion fruit (Passiflora edulis) is a perennial evergreen vine that grows mainly in tropical and subtropical regions due to its nutritional, medicinal and ornamental values. However, the molecular biology study of passion fruit is extremely hindered by the lack of an easy and efficient method for transformation. The protoplast transformation system plays a vital role in plant regeneration, gene function analysis and genome editing. Here, we present a new method ('Cotyledon Peeling Method') for simple and efficient passion fruit protoplast isolation using cotyledon as the source tissue. A high yield (2.3 × 107 protoplasts per gram of fresh tissues) and viability (76%) of protoplasts were obtained upon incubation in the enzyme solution [1% (w/v) cellulase R10, 0.25% (w/v) macerozyme R10, 0.4 M mannitol, 10 mM CaCl2, 20 mM KCl, 20 mM MES and 0.1% (w/v) BSA, pH 5.7] for 2 hours. In addition, we achieved high transfection efficiency of 83% via the polyethylene glycol (PEG)-mediated transformation with a green fluorescent protein (GFP)-tagged plasmid upon optimization. The crucial factors affecting transformation efficiency were optimized as follows: 3 µg of plasmid DNA, 5 min transfection time, PEG concentration at 40% and protoplast density of 100 × 104 cells/ml. Furthermore, the established protoplast system was successfully applied for subcellular localization analysis of multiple fluorescent organelle markers and protein-protein interaction study. Taken together, we report a simple and efficient passion fruit protoplast isolation and transformation system, and demonstrate its usage in transient gene expression for the first time in passion fruit. The protoplast system would provide essential support for various passion fruit biology studies, including genome editing, gene function analysis and whole plant regeneration.

Structures of sperm flagellar doublet microtubules expand the genetic spectrum of male infertility.

Sperm motility is crucial for successful fertilization. Highly decorated doublet microtubules (DMTs) form the sperm tail skeleton, which propels the movement of spermatozoa. Using cryo-electron microscopy (cryo-EM) and artificial intelligence (AI)-based modeling, we determined the structures of mouse and human sperm DMTs and built an atomic model of the 48-nm repeat of the mouse sperm DMT. Our analysis revealed 47 DMT-associated proteins, including 45 microtubule inner proteins (MIPs). We identified 10 sperm-specific MIPs, including seven classes of Tektin5 in the lumen of the A tubule and FAM166 family members that bind the intra-tubulin interfaces. Interestingly, the human sperm DMT lacks some MIPs compared with the mouse sperm DMT. We also discovered variants in 10 distinct MIPs associated with a subtype of asthenozoospermia characterized by impaired sperm motility without evident morphological abnormalities. Our study highlights the conservation and tissue/species specificity of DMTs and expands the genetic spectrum of male infertility.

A Zinc Finger Motif in the P1 N Terminus, Highly Conserved in a Subset of Potyviruses, Is Associated with the Host Range and Fitness of Telosma Mosaic Virus.

P1 is the first protein translated from the genomes of most viruses in the family Potyviridae, and it contains a C-terminal serine-protease domain that cis-cleaves the junction between P1 and HCPro in most cases. Intriguingly, P1 is the most divergent among all mature viral factors, and its roles during viral infection are still far from understood. In this study, we found that telosma mosaic virus (TelMV, genus Potyvirus) in passion fruit, unlike TelMV isolates present in other hosts, has two stretches at the P1 N terminus, named N1 and N2, with N1 harboring a Zn finger motif. Further analysis revealed that at least 14 different potyviruses, mostly belonging to the bean common mosaic virus subgroup, encode a domain equivalent to N1. Using the newly developed TelMV infectious cDNA clones from passion fruit, we demonstrated that N1, but not N2, is crucial for viral infection in both Nicotiana benthamiana and passion fruit. The regulatory effects of N1 domain on P1 cis cleavage, as well as the accumulation and RNA silencing suppression (RSS) activity of its cognate HCPro, were comprehensively investigated. We found that N1 deletion decreases HCPro abundance at the posttranslational level, likely by impairing P1 cis cleavage, thus reducing HCPro-mediated RSS activity. Remarkably, disruption of the Zn finger motif in N1 did not impair P1 cis cleavage and HCPro accumulation but severely debilitated TelMV fitness. Therefore, our results suggest that the Zn finger motif in P1s plays a critical role in viral infection that is independent of P1 protease activity and self-release, as well as HCPro accumulation and silencing suppression. IMPORTANCE Viruses belonging to the family Potyviridae represent the largest group of plant-infecting RNA viruses, including a variety of agriculturally and economically important viral pathogens. Like all picorna-like viruses, potyvirids employ polyprotein processing as the gene expression strategy. P1, the first protein translated from most potyvirid genomes, is the most variable viral factor and has attracted great scientific interest. Here, we defined a Zn finger motif-encompassing domain (N1) at the N terminus of P1 among diverse potyviruses phylogenetically related to bean common mosaic virus. Using TelMV as a model virus, we demonstrated that the N1 domain is key for viral infection, as it is involved both in regulating the abundance of its cognate HCPro and in an as-yet-undefined key function unrelated to protease processing and RNA silencing suppression. These results advance our knowledge of the hypervariable potyvirid P1s and highlight the importance for infection of a previously unstudied Zn finger domain at the P1 N terminus.

Structure of a TOC-TIC supercomplex spanning two chloroplast envelope membranes.

The TOC and TIC complexes are essential translocons that facilitate the import of the nuclear genome-encoded preproteins across the two envelope membranes of chloroplast, but their exact molecular identities and assembly remain unclear. Here, we report a cryoelectron microscopy structure of TOC-TIC supercomplex from Chlamydomonas, containing a total of 14 identified components. The preprotein-conducting pore of TOC is a hybrid ß-barrel co-assembled by Toc120 and Toc75, while the potential translocation path of TIC is formed by transmembrane helices from Tic20 and YlmG, rather than a classic model of Tic110. A rigid intermembrane space (IMS) scaffold bridges two chloroplast membranes, and a large hydrophilic cleft on the IMS scaffold connects TOC and TIC, forming a pathway for preprotein translocation. Our study provides structural insights into the TOC-TIC supercomplex composition, assembly, and preprotein translocation mechanism, and lays a foundation to interpret the evolutionary conservation and diversity of this fundamental translocon machinery.

Investigation of transverse crack spacing in an asphalt pavement with a semi-rigid base.

Transverse cracking is a serious problem for semi-rigid base asphalt pavement. The shrinkage of the base course and the surface course, as well as reflective cracks, are key factors for transverse cracking in asphalt pavement. Crack spacing can directly reflect the degree of transverse cracking in pavements. Therefore, this study aims to calculate the transverse crack spacing and discuss its affecting factors. To this end, a calculation model of transverse crack spacing for the semi-rigid base asphalt pavement was first established. Then, the transverse crack spacings of different composite structures were calculated, and the influences of the shrinkage coefficient, the structural layer thickness, and the pavement tensile strength on transverse crack spacing were expounded. Finally, the transverse crack spacing of the pavement after the appearance of the reflective was calculated. The results show that the lower lime and fly ash content and skeleton gap gradation can be adopted during the design of the base course. Meanwhile, the lower lime and fly ash content in the macadam base, the skeleton gap gradation and asphalt concrete with a larger particle size in the surface layer can be used during the design of surface layer. In addition, the transverse crack spacing of the semi-rigid base asphalt pavement could be increased by reducing the shrinkage coefficient, increasing the thicknesses of the surface course and the base course, and improving the tensile strength of the pavement. After the appearance of reflective cracks, the transverse crack spacing of the surface layer ranged between 32.8 m and 66.5 m. 15fp-AC25, 15fp-AC20, 15df-AC25, and 17fp-AC25 were found to be the best semi-rigid base asphalt pavement structures to reduce transverse cracking. Finally, transverse cracking in pavement composite structures under different bonding conditions needs to be analyzed in the follow-up work.

Effects of Tung Oil Composite Regenerating Agent on Rheological Properties and Microstructures of Reclaimed Asphalt Binder.

The single light oil regenerating agent has certain limitations on the performance recovery of aged asphalt. In this study, tung oil, dioctyl phthalate (DOP), C9 petroleum resin, and organic montmorillonite (OMMT) were used to prepare the composite regenerating agent, and its optimal mix proportion was determined by the orthogonal experimental design. The rheological properties and anti-aging performance of reclaimed asphalt were studied by the dynamic shear rheometer (DSR) and bending beam rheometer (BBR); and the Fourier transform infrared (FTIR) spectrometer, gel permeation chromatography (GPC), and scanning electron microscope (SEM) were adopted to explore its microstructure, morphology, and mechanism of action. The results show that with the addition of tung oil composite regenerating agent, the rheological properties of aged asphalt can be effectively recovered, even better than that of base asphalt. By using the complex modulus aging index (CMAI) and phase angle aging index (PMAI) it is found that the anti-aging performance of reclaimed asphalt is better than that of base asphalt. With the optimal content of the tung oil composite regenerating agent, the contents of characteristic functional groups and macromolecular asphaltenes in the aged asphalt can be reduced, indicating that the composite regenerating agent is beneficial to the dispersion and dissolution of polar substances in the aged asphalt. After aging, a large number of wrinkles appear on the surface of the asphalt. However, the addition of the tung oil composite regenerating agent can make the asphalt surface smooth, which indicates that the tung oil composite regenerating agent can restore the microstructure and morphology of aged asphalt to a certain extent.

Architecture of the human NALCN channelosome.

NALCN regulates the resting membrane potential by mediating the Na+ leak current in neurons, and it functions as a channelosome in complex with FAM155A, UNC79, and UNC80. Dysfunction of the NALCN channelosome causes a broad range of neurological and developmental diseases called NALCN channelopathies in humans. How the auxiliary subunits, especially the two large components UNC79 and UNC80, assemble with NALCN and regulate its function remains unclear. Here we report an overall architecture of the human NALCN channelosome. UNC79 and UNC80 each adopt an S-shape super-helical structure consisting of HEAT and armadillo repeats, forming a super-coiled heterodimeric assembly in the cytoplasmic side, which may provide a scaffold for the binding of other potential modulators of the channelosome. The UNC79-UNC80 assembly specifically associates with the NALCN-FAM155A subcomplex through the intracellular II-III linker of NALCN. Disruptions of the interaction interfaces between UNC79 and UNC80, and between the II-III linker of NALCN and the UNC79-UNC80 assembly, significantly reduce the NALCN-mediated currents in HEK293T system, suggesting the importance of the UNC79-UNC80 assembly in regulating channelosome function. Cross-linking mass spectrometry analysis identified an additional calmodulin (CaM) bound in the carboxyl-terminal domain of NALCN. Our study thus provides a structural basis for understanding the unique assembly mechanism and functional regulation of the NALCN channelosome, and also provides an opportunity for the interpretation of many disease-related mutations in UNC80.

NiCo2O4 spinel for efficient toluene oxidation: The effect of crystal plane and solvent.

Spinel oxides, e.g., NiCo2O4, is a promising catalyst for the catalytic oxidation of toluene. Understanding and designing versatile NiCo2O4 spinel is important for low-temperature toluene oxidation. Here, we investigated the surface-characteristic-dependent degradation activity of NiCo2O4 crystals through experiment and characterization. NiCo2O4 nanosheet using ethanol as solvent (named E--NiCo2O4) exposing {110} crystal planes exhibited the lowest temperature toluene oxidation. The T99 of toluene conversion was 256 °C, which is much lower than that of NiCo2O4 nanosheet using ethylene glycol as solvent (named EG--NiCo2O4), NiCo2O4 octahedron (named O--NiCo2O4) and NiCo2O4 truncated octahedron (named TO--NiCo2O4). Characterization using various techniques such as XRD, TEM, BET, XPS, H2-TPR and CO2-TPD showed that Co3+ and surface adsorbed oxygen (Osur) enriched surface, excellent redox properties and effective diffusion of the reaction product reasonably explain the enhancement in catalytic activity over the E--NiCo2O4. The research reveals that the effect of specific crystal planes and solvent was the key factor to govern the activity of low-temperature toluene oxidation.

Preparation of metal-doped Cu-Mn/HTS-1 catalysts and their mechanisms in efficient degradation of toluene.

In order to study their synergistic catalytic effects in toluene degradation, CuMn2O4/HTS-1 (HTS-1 was a titanium silicon molecular sieve), Cu0.7Mn2Y0.3Ox/HTS-1 and Cu0.7Mn2Ce0.3Ox/HTS-1 catalysts were prepared by the impregnation method. The textural properties, redox properties and acidity of the catalysts were characterized by X-ray diffractometer (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), H2 temperature-programmed reduction (H2-TPR), X-ray photoelectron spectroscopy (XPS), frustrated total internal reflection (FT-IR), ammonium temperature-programmed desorption (NH3-TPD) and pyridine adsorption internal reflection (Py-IR) measurements. The potential roles of Lewis acid sites (activating dioxygen) were discussed, and the experimental results indicated that the most efficient route for toluene degradation over Cu0.7Mn2Ce0.3Ox/HTS-1 (toluene conversion rate of 90% (T99)=295°C) was ascribed to regulation of the synergistic effects of redox properties (activating molecular toluene) and Lewis acid sites (activating dioxygen). The Mars-Van-Krevelen (MVK) model was adopted to describe the reaction process of toluene oxidation, which gave an in-depth view into the toluene degradation over CuMn2O4/HTS-1, Cu0.7Mn2Y0.3Ox/HTS-1 and Cu0.7Mn2Ce0.3Ox/HTS-1. In addition, the synergistic effects between redox properties and Lewis acid sites were studied in detail.

Process and mechanism of toluene oxidation using Cu1-yMn2CeyOx/sepiolite prepared by the co-precipitation method.

To achieve efficient degradation of toluene, a series of Cu1-yMn2CeyOx/sepiolite catalysts (y = 0.1, 0.2, and 0.3) with different Cu1-yMn2CeyOx loadings (10%, 20%, and 30%) were prepared via the co-precipitation method. The structure-activity and surficial elemental species of Cu1-yMn2CeyOx/sepiolite were characterized by XRD, TEM, SEM, BET, ICP-MS and XPS. The catalytic activity of the catalysts was tested in the oxidation reaction of toluene, results showed that 20%Cu0.8Mn2Ce0.2Ox/sepiolite remains able to remove toluene completely with high efficiency at a temperature of 289 °C. Two kinetic models have been selected and tested to describe the oxidation of toluene, the Mars-van krevele (MVK) model provided a good fit (R2 ≥ 0.99). And the optimal relation of the surface oxidation activation energy (26.074 kJ mol-1) and surface reduction activation energy (23.591 kJ mol-1) were calculated.