Quantitative Isothermal Amplification on Paper Membranes using Amplification Nucleation Site Analysis

Quantitative nucleic acid amplification tests (qNAATs) are critical in treating infectious diseases, such as in HIV viral load monitoring or SARS-CoV-2 testing, in which viral load indicates viral suppression or infectivity. Quantitative PCR is the gold standard tool for qNAATs; however, there is a need to develop point-of-care (POC) qNAATs to manage infectious diseases in outpatient clinics, low- and middle-income countries, and the home. Isothermal amplification methods are an emerging tool for POC NAATs as an alternative to traditional PCR-based workflows. Previous works have focused on relating isothermal amplification bulk fluorescence signals to input copies of target nucleic acids for sample quantification with limited success. In this work, we show that recombinase polymerase amplification (RPA) reactions on paper membranes exhibit discrete fluorescent amplification nucleation sites. We demonstrate that the number of nucleation sites can be used to quantify HIV-1 DNA and RNA in less than 20 minutes. An image-analysis algorithm quantifies nucleation sites and determines the input nucleic acid copies in the range of 67-3,000 copies per reaction. We demonstrate a mobile phone-based system for image capture and onboard processing, illustrating that this method may be used at the point-of-care for qNAATs with minimal instrumentation.

The Impact of Nano Clay on Normal and High-Performance Concrete Characteristics: A Review

The use of nano clay to improve the qualities of construction materials and engineering applications has attracted a lot of discussion in recent years. This review article summarizes the influence of nano clay as a cement substitute and supplement on the performance of conventional and high-performance concrete. The addition of nano clay to high performance concrete revealed an increase in compressive and flexural strength, as well as durability attributes such as resistance to elevated temperatures and sulfate attack, while simultaneously decreasing porosity, permeability, and water absorption. This enhancement is a result of nano clay’s roles as nano reinforcements, nanofillers, nucleation sites, and reactive pozzolans, which promote hydration and increase material characteristics.

High-yield synthesis of CsPbBr3 nanoparticles: Diphenylphosphine as a reducing agent and its effect in Pb-seeding nucleation and growth

Based on the reported nucleation mechanisms for CsPbX3 and II-VI/IV-VI quantum dots, CsPbBr3 nanoparticles with a high reaction-yield, up to 393% mass-increment, were synthesized by the hot-injection method. The introduction of diphenylphosphine (DPP) as a reducing agent improved nanoparticle nucleation and growth, giving out evidence for Pb-seeding in CsPbBr3 nanoparticles formation. Additionally, a clear influence of the DPP in a CsPbBr3-Cs4PbBr6 incomplete phase transformation was observed, marked by the appearance of several PbBr2 nanoparticles, indicating the need for an improved ratio between the stabilizing agents and the precursors, due to the increased number of nucleation sites produced by the DPP. The resulting CsPbBr3 nanoparticles showed high quality, as they displayed 70%-90% photoluminescence quantum yield (PLQY), narrow size distribution with an average nanoparticle size of ~10 nm and the characteristic cubic morphology reported in previous works. This increment in CsPbBr3 nanoparticles’ reaction yield will contribute to making them a more attractive option for different optoelectronic applications.

Structure and Magnetic Properties of Thermodynamically Predicted Rapidly Quenched Fe85-xCuxB15 Alloys

In this work, based on the thermodynamic prediction, the comprehensive studies of the influence of Cu for Fe substitution on the crystal structure and magnetic properties of the rapidly quenched Fe85B15 alloy in the ribbon form are performed. Using thermodynamic calculations, the parabolic shape dependence of the ΔGamoprh with a minimum value at 0.6% of Cu was predicted. The ΔGamoprh from the Cu content dependence shape is also asymmetric, and, for Cu = 0% and Cu = 1.5%, the same ΔGamoprh value is observed. The heat treatment optimization process of all alloys showed that the least lossy (with a minimum value of core power losses) is the nanocomposite state of nanocrystals immersed in an amorphous matrix obtained by annealing in the temperature range of 300–330 °C for 20 min. The minimum value of core power losses P10/50 (core power losses at 1T@50Hz) of optimally annealed Fe85-xCuxB15 x = 0,0.6,1.2% alloys come from completely different crystallization states of nanocomposite materials, but it strongly correlates with Cu content and, thus, a number of nucleation sites. The TEM observations showed that, for the Cu-free alloy, the least lossy crystal structure is related to 2–3 nm short-ordered clusters; for the Cu = 0.6% alloy, only the limited value of several α-Fe nanograins are found, while for the Cu-rich alloy with Cu = 1.2%, the average diameter of nanograins is about 26 nm, and they are randomly distributed in the amorphous matrix. The only high number of nucleation sites in the Cu = 1.2% alloy allows for a sufficient level of grains’ coarsening of the α-Fe phase that strongly enhances the ferromagnetic exchange between the α-Fe nanocrystals, which is clearly seen with the increasing value of saturation induction up to 1.7T. The air-annealing process tested on studied alloys for optimal annealing conditions proves the possibility of its use for this type of material.

The metadynamic recrystallization behavior of ultrahigh-strength stainless steel: effects of precipitates and shear bands

The metadynamic recrystallization behavior of Cr-Co-Ni-Mo ultrahigh-strength martensitic stainless steel was studied in a double-pass isothermal compression test, and a metadynamic recrystallization kinetics model for softening was established. The results showed that the metadynamic recrystallization softening rate of the steel not only depended on the deformation temperature and strain rate but was also related to the dynamic precipitation and the local shear bands in the steel. When the deformation temperature was below 1050 °C, the dynamically precipitated M6C carbides pinned the grain boundaries and hindered metadynamic recrystallization. When the steel was deformed at a deformation temperature of 1000~1050 °C and a strain rate of 1.0~5.0 s-1, a large number of local shear bands were generated. The local shear bands increased the number of nucleation sites for dynamic recrystallization and enhanced the softening rate of metadynamic recrystallization.

Microcrystalline cellulose effects on the rheology of mixed oleogels structured with candelilla wax and saturated fat

The structuration processes of mixed oleogels produced with candelilla wax (CW, 0 or 3%), fully hydrogenated soybean oil (FH, 5-15%), and microcrystalline cellulose (MC, 0-9%) were studied to define their rheological effects. During the cooling CW crystals performed as nucleation sites for FH. The elastic modulus (G’) of oleogels with FH and 3% CW were more than two orders of magnitude higher than those produced with 0% CW. Adding MC to the oleogels increased slightly the G’. Independently of the amount of MC, oleogels structured with increasing amounts of FH and 0% CW showed the elastic properties scaling of colloidal gels. This behavior was lost by adding 3% CW, implying that in mixed FH-CW oleogels, the CW crystal network dominated the oleogel rheology. The flow point and the mechanical reversibility of oleogels and commercial butter (CB) was also determined. CB showed flow points at 44 and 59% strain and mechanical reversibility values of 29 and 35% of G’ measured in a pre-shear step. Adding MC to oleogels structured with FH and 0% CW increased their flow point (37.2%) near those of CB. This effect was not produced in mixed FH-3% CW oleogels. The mechanical recovery of oleogels produced with FH, MC, and 0% CW tend to decrease as the FH content increased. CW and MC did not show a simple concentration–effect relationship for the mechanical recovery. Nonetheless, oleogels structured with 3% CW and 10% FH and 6-9% MC showed mechanical recovery (~60%) close to that of CB.

Multiwalled carbon nanotubes synthesis from methane using a stainless steel foils as a catalyst

In this study, a thin stainless-steel foil was used as a catalyst for carbon nanotubes (CNTs) using methane as a carbon source via the chemical vapor deposition (CVD) method. Our results revealed that pre-treatment step of the catalyst plays an important role in CNT formation. In our experiments, a catalyst pre-treatment temperature of 850 oC have been found to facilitate the surface roughness and provide more active nucleation sites for CNTs formation. Multiwalled CNTs with 6 layers, their diameters of 10 – 20 nm and their length of app. 300 nm were grown. This finding might lead to a process for improving the quality of MWCNTs grown on steel foil as catalyst.

Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis

Background Microbially induced calcite precipitation (MICP) is an ancient property of bacteria, which has recently gained considerable attention for biotechnological applications. It occurs as a by-product of bacterial metabolism and involves a combination of chemical changes in the extracellular environment, e.g. pH increase, and presence of nucleation sites on the cell surface or extracellular substances produced by the bacteria. However, the molecular mechanisms underpinning MICP and the interplay between the contributing factors remain poorly understood, thus placing barriers to the full biotechnological and synthetic biology exploitation of bacterial biomineralisation. Results In this study, we adopted a bottom-up approach of systematically engineering Bacillus subtilis, which has no detectable intrinsic MICP activity, for biomineralisation. We showed that heterologous production of urease can induce MICP by local increases in extracellular pH, and this can be enhanced by co-expression of urease accessory genes for urea and nickel uptake, depending on environmental conditions. MICP can be strongly enhanced by biofilm-promoting conditions, which appeared to be mainly driven by production of exopolysaccharide, while the protein component of the biofilm matrix was dispensable. Attempts to modulate the cell surface charge of B. subtilis had surprisingly minor effects, and our results suggest this organism may intrinsically have a very negative cell surface, potentially predisposing it for MICP activity. Conclusions Our findings give insights into the molecular mechanisms driving MICP in an application-relevant chassis organism and the genetic elements that can be used to engineer de novo or enhanced biomineralisation. This study also highlights mutual influences between the genetic drivers and the chemical composition of the surrounding environment in determining the speed, spatial distribution and resulting mineral crystals of MICP. Taken together, these data pave the way for future rational design of synthetic precipitator strains optimised for specific applications.