Development of a Rapid Fluorescent Diagnostic System for Early Detection of the Highly Pathogenic Avian Influenza H5 Clade 2.3.4.4 Viruses in Chicken Stool

Rapid diagnosis is essential for the control and prevention of H5 highly pathogenic avian influenza viruses (HPAIVs). However, highly sensitive and rapid diagnostic systems have shown limited performance due to specific antibody scarcity. In this study, two novel specific monoclonal antibodies (mAbs) for clade 2.3.4.4 H5Nx viruses were developed by using an immunogen from a reversed genetic influenza virus (RGV). These mAbs were combined with fluorescence europium nanoparticles and an optimized lysis buffer, which were further used for developing a fluorescent immunochromatographic rapid strip test (FICT) for early detection of H5Nx influenza viruses on chicken stool samples. The result indicates that the limit of detection (LoD) of the developed FICT was 40 HAU/mL for detection of HPAIV H5 clade 2.3.4.4b in spiked chicken stool samples, which corresponded to 4.78 × 10⁴ RNA copies as obtained from real-time polymerase chain reaction (RT-PCR). An experimental challenge of chicken with H5N6 HPAIV is lethal for chicken three days post-infection (DPI). Interestingly, our FICT could detect H5N6 in stool samples at 2 DPI earlier, with 100% relative sensitivity in comparison with RT-PCR, and it showed 50% higher sensitivity than the traditional colloidal gold-based rapid diagnostic test using the same mAbs pair. In conclusion, our rapid diagnostic method can be utilized for the early detection of H5Nx 2.3.4.4 HPAIVs in avian fecal samples from poultry farms or for influenza surveillance in wild migratory birds.     

Treatment of Mouse Sperm with a Non-Catalytic Mutant of PLA2G10 Reveals That PLA2G10 Improves In Vitro Fertilization through Both Its Enzymatic Activity and as Ligand of PLA2R1

The group X secreted phospholipase A2 (PLA2G10) is present at high levels in mouse sperm acrosome. The enzyme is secreted during capacitation and amplifies the acrosome reaction and its own secretion via an autocrine loop. PLA2G10 also improves the rate of fertilization. In in vitro fertilization (IVF) experiments, sperm from Pla2g10-deficient mice produces fewer two-cell embryos, and the absence of PLA2G10 is rescued by adding recombinant enzymes. Moreover, wild-type (WT) sperm treated with recombinant PLA2G10 produces more two-cell embryos. The effects of PLA2G10 on mouse fertility are inhibited by sPLA₂ inhibitors and rescued by products of the enzymatic reaction such as free fatty acids, suggesting a role of catalytic activity. However, PLA2G10 also binds to mouse PLA2R1, which may play a role in fertility. To determine the relative contribution of enzymatic activity and PLA2R1 binding in the profertility effect of PLA2G10, we tested H48Q-PLA2G10, a catalytically-inactive mutant of PLA2G10 with low enzymatic activity but high binding properties to PLA2R1. Its effect was tested in various mouse strains, including Pla2r1-deficient mice. H48Q-PLA2G10 did not trigger the acrosome reaction but was as potent as WT-PLA2G10 to improve IVF in inbred C57Bl/6 mice; however, this was not the case in OF1 outbred mice. Using gametes from these mouse strains, the effect of H48Q-PLA2G10 appeared dependent on both spermatozoa and oocytes. Moreover, sperm from C57Bl/6 Pla2r1-deficient mice were less fertile and lowered the profertility effects of H48Q-PLA2G10, which were completely suppressed when sperm and oocytes were collected from Pla2r1-deficient mice. Conversely, the effect of WT-PLA2G10 was not or less sensitive to the absence of PLA2R1, suggesting that the effect of PLA2G10 is polymodal and complex, acting both as an enzyme and a ligand of PLA2R1. This study shows that the action of PLA2G10 on gametes is complex and can simultaneously activate the catalytic pathway and the PLA2R1-dependent receptor pathway. This work also shows for the first time that PLA2G10 binding to gametes’ PLA2R1 participates in fertilization optimization.     

Absence of CCR2 Promotes Proliferation of Alveolar Macrophages That Control Lung Inflammation in Acute Respiratory Distress Syndrome in Mice

Acute respiratory distress syndrome (ARDS) consists of uncontrolled inflammation that causes hypoxemia and reduced lung compliance. Since it is a complex process, not all details have been elucidated yet. In a well-controlled experimental murine model of lipopolysaccharide (LPS)-induced ARDS, the activity and viability of macrophages and neutrophils dictate the beginning and end phases of lung inflammation. C-C chemokine receptor type 2 (CCR2) is a critical chemokine receptor that mediates monocyte/macrophage activation and recruitment to the tissues. Here, we used CCR2-deficient mice to explore mechanisms that control lung inflammation in LPS-induced ARDS. CCR2^−/− mice presented higher total numbers of pulmonary leukocytes at the peak of inflammation as compared to CCR2^+/+ mice, mainly by enhanced influx of neutrophils, whereas we observed two to six-fold lower monocyte or interstitial macrophage numbers in the CCR2^−/−. Nevertheless, the time needed to control the inflammation was comparable between CCR2^+/+ and CCR2^−/−. Interestingly, CCR2^−/− mice presented higher numbers and increased proliferative rates of alveolar macrophages from day 3, with a more pronounced M2 profile, associated with transforming growth factor (TGF)-β and C-C chemokine ligand (CCL)22 production, decreased inducible nitric oxide synthase (Nos2), interleukin (IL)-1β and IL-12b mRNA expression and increased mannose receptor type 1 (Mrc1) mRNA and CD206 protein expression. Depletion of alveolar macrophages significantly delayed recovery from the inflammatory insult. Thus, our work shows that the lower number of infiltrating monocytes in CCR2^−/− is partially compensated by increased proliferation of resident alveolar macrophages during the inflammation control of experimental ARDS.     

Identification and Characterization of Malate Dehydrogenases in Tomato (Solanum lycopersicum L.)

Malate dehydrogenase, which facilitates the reversible conversion of malate to oxaloacetate, is essential for energy balance, plant growth, and cold and salt tolerance. However, the genome-wide study of the MDH family has not yet been carried out in tomato (Solanum lycopersicum L.). In this study, 12 MDH genes were identified from the S. lycopersicum genome and renamed according to their chromosomal location. The tomato MDH genes were split into five groups based on phylogenetic analysis and the genes that clustered together showed similar lengths, and structures, and conserved motifs in the encoded proteins. From the 12 tomato MDH genes on the chromosomes, three pairs of segmental duplication events involving four genes were found. Each pair of genes had a Ka/Ks ratio < 1, indicating that the MDH gene family of tomato was purified during evolution. Gene expression analysis exhibited that tomato MDHs were differentially expressed in different tissues, at various stages of fruit development, and differentially regulated in response to abiotic stresses. Molecular docking of four highly expressed MDHs revealed their substrate and co-factor specificity in the reversible conversion process of malate to oxaloacetate. Further, co-localization of tomato MDH genes with quantitative trait loci (QTL) of salt stress-related phenotypes revealed their broader functions in salt stress tolerance. This study lays the foundation for functional analysis of MDH genes and genetic improvement in tomato.     

Mechanical Properties of Highly Deformable Elastomeric Gyroids for Multifunctional Capacitors

Triply periodic minimal surface lattices have mechanical properties that derive from the unit cell geometry and the base material. Through computation software like nTopology and Abaqus, these geometries are used to tune nonlinear stress–strain curves not readily achievable with solid materials alone and to change the compliance by two orders of magnitude compared to the constituent material. In this study, four elastomeric TPMS gyroids undergo large deformation compression and tension testing to investigate the impact of the structure's geometry on the mechanical properties. Among all the samples, the modulus at strain ε$=0.05$ varies by over one order of magnitude (7.7–293.4 kPa from FEA under compression). These lattices are promising candidates for designing multifunctional systems that can perform multiple tasks simultaneously by leveraging the geometry's large surface area to volume ratio. For example, the architectural functionality of the lattice to bear loads and store mechanical energy along with the larger surface area for energy storage is combined. A compliant double-gyroid capacitor that can simultaneously achieve three functions is demonstrated: load bearing, energy storage, and sensing.     

CO2 utilization in Eastern Canada: Sources,sites, and grid effects

Achieving net zero carbon dioxide (CO₂) emissions will require the cessation of fossil fuel emissions into the atmosphere, yet the need for ‘fuel’ and energy storage will remain. One solution could be a carbon-based fuel system where CO₂ of biogenic origin is converted to fuels using hydrogen generated by electrolysis powered by renewable energy sources. Methane has value as an initial target given its prevalence in biogas, use in home heating and in electricity generation. Sources of CO₂ in Eastern Canada are dominated by the iron and steel, cement, and aluminium industries, all of which have biogenic fuel options. Collecting all of the potentially biogenic CO₂ would displace 75% of current natural gas use and require a 50% increase in generating capacity. Initial efforts could site a carbon capture, utilization, and storage facility near Montreal, QC, with other large-scale facilities near Hamilton, ON, and Lac St-Jean, QC. These facilities would be grid connected and expected to operate ~6200 h annually. The most high-frequency electrolysis events would be 10 h of run time and 2 h of idle time. These periods would peak during the equinox months and be at a minimum during the winter solstice. These operational assumptions will all be subject to the increased variability caused by anthropogenic climate change and increased renewable generation on the grid. A closed-loop carbon-based fuel system would require an equivalent price of $250 per tonne CO₂.     

Mechanical and microstructural investigation of dissimilar joints of Al-Cu and Cu-Al metals using nanosecond laser

Laser beam welding (LBW) has found wide applications in several fields, including electronics, aerospace, and automobile industries, owing to its inherent capabilities over existing welding techniques. Dissimilar laser welding is one of the challenging fabrication processes. However, joining of different materials poses challenging issues because material properties of each material interacts to give rise to hybrid system performance. In present study weld of Al-Cu and Cu-Al has been performed using Ytterbium pulsed fibre laser with a maximum laser power of 300 W. The scanning electron microscope (SEM), and energy dispersive x-ray spectroscopy (EDS) has been employed to investigate the microstructural and chemical composition of the welded joints. The EDS analysis showed the presence of possible intermetallic compounds (IMCs) like AlCu and Cu2Al in the fusion zone. The Al-Cu and Cu-Al welded joints had maximum shear strength of 295 N and 84 N, respectively. The results showed the efficacy of Al-Cu joints over Cu-Al joints. The study further demonstrated that the Al-Cu weld had a better microstructure with fewer pores and cracks than the Cu-Al welds by varying the laser powers.

     

Effect of climatic factors on farm income of rain-fed and irrigated farming households in Vietnam

The paper provides new evidence of the effect of temperature and precipitation on crop profit of farmers applying different farming systems across different seasons as well as geographical regions in Vietnam for the period 2004–2016. The result finds that the effect of fluctuation in temperature and rainfall on the crop profit of farmers is not uniform across the three regions (North, Central and South) and also considerable variation across rain-fed and irrigated farms. For the rain-fed farming system, (i) rainfall in the dry season has a nonlinearly (⋂) association with the profit, but rainfall in the rainy season has no effect, and (ii) dry season temperature and rainy season temperature have, respectively, ⋂-shaped and U-shaped relationship with the profit. For the irrigated farming systems, the temperatures in the dry season and rainy season have a U-shaped relationship with the profit. The findings show that the irrigated farming systems have been effective in the long term in the face of climate change. Therefore, in preparation for adaptation to climate change, the Vietnamese agricultural sector needs a complete irrigation system at both farm and regional levels.