Isothermal Molecular Dynamics Simulations Research Articles

Insight into the interaction of serum albumin with antihypertensive peptide Val-Ala-Pro from bovine casein hydrolysate based on the biolayer interferometry, multi-spectroscopic analysis and computational evaluation

Food-derived angiotensin-converting enzyme inhibitory peptide (ACEIP) has an effect in supportive therapeutic on hypertension. Bovine serum albumin (BSA) as a model transporter protein to explore the interaction mechanisms with casein-hydrolyzed ACEIP Val-Ala-Pro (VAP) by multi-spectroscopic, biolayer interferometry (BLI), isothermal titration calorimetry (ITC), molecular docking, and molecular dynamics simulations. Multi-spectroscopic analysis showed that the non-covalent complexes formed by VAP and BSA resulted in decreased hydrophobicity and α-helix contents on BSA, revealing the unfolding of the BSA structure. BLI revealed the reversible binding process of BSA to VAP. ITC confirmed that the combination of VAP to BSA was a spontaneous process mainly driven by entropy. Molecular docking and molecular dynamic simulations showed that VAP was primarily bound in site II of BSA by hydrogen bonding, hydrophobic interactions, van der Waals force, and electrostatic force. This study provides a systematic method to reveal the structure–activity relationship of ACEIPs.

The AD3 locus of synaptotagmin-1 C2 domains modulates domain stability

Synaptotagmin-1 (syt1) functions as the Ca2+-dependent sensor that triggers the rapid and synchronous release of neurotransmitters from neurotransmitter-containing vesicles during neuronal exocytosis. The syt1 protein has two homologous tandem C2 domains that interact with phospholipids in a Ca2+-dependent manner. Despite the crucial role of syt1 in exocytosis, the precise interactions between Ca2+, syt1, and phospholipids are not fully understood. In a study involving recessive lethal mutations in the syt1 gene, a specific mutation named AD3 was generated in Drosophila syt1, resulting in a significant reduction in Ca2+-dependent exocytosis. Further investigation revealed that the AD3 mutation was a missense mutation located in a conserved consensus sequence within the C2B domain of Drosophila syt1. However, the biophysical impact of the AD3 mutation had not been analyzed. Our study uses x-ray crystallography, isothermal titration calorimetry, thermodynamic analysis, and molecular dynamics simulation to show that the primary defect caused by the AD3 mutation in the syt1 protein is reduced thermodynamic stability. This instability alters the population of Ca2+-receptive states, leading to two major consequences: decreased affinity for calcium ions and compromised stabilization of the domain normally enhanced by Ca2+. We conclude that this conserved residue acts as a structural constraint, delimiting the movement of loop 3 within the pocket and ultimately influencing the affinity of the calcium ion binding with the C2 domain.

Destabilization of the D2 domain of Thermotoga maritima arginine binding protein induced by guanidinium thiocyanate and its counteraction by stabilizing agents.

D2 is a structural and cooperative domain of Thermotoga maritima Arginine Binding Protein, that possesses a remarkable conformational stability, with a denaturation temperature of 102.6°C, at pH 7.4. The addition of potassium thiocyanate causes a significant decrease in the D2 denaturation temperature. The interactions of thiocyanate ions with D2 have been studied by means of isothermal titration calorimetry measurements and molecular dynamics simulations. It emerged that: (a) 20-30 thiocyanate ions interact with the D2 surface and are present in its first solvation shell; (b) each of them makes several contacts with protein groups, both polar and nonpolar ones. The addition of guanidinium thiocyanate causes a marked destabilization of the D2 native state, because both the ions are denaturing agents. However, on adding to the solution containing D2 and guanidinium thiocyanate a stabilizing agent, such as TMAO, sucrose or sodium sulfate, a significant increase in denaturation temperature occurs. The present results confirm that counteraction is a general phenomenon for globular proteins.

Preliminary exploration of the anti-ovarian cancer activity of peptides derived from bovine bone collagen hydrolysate and its related mechanisms

Ovarian cancer, a malignant tumor that poses a significant threat to women's health, has seen a rise in incidence, prompting the urgent need for more effective treatment. This study primarily aimed to explore the potential of bovine collagen peptides in inhibiting ovarian cancer. The investigation in this study began with the identification of 268 peptide sequences through LC-MS/MS, followed by a screening process using molecular docking techniques to identify potential peptides capable of binding to EGFR. Subsequently, a series of experiments were performed, demonstrating the inhibitory effects of the peptide GPAGADGDRGEAGPAGPAGPAGPR on the proliferation of ovarian cancer cells. Transcriptomic analysis further revealed that this peptide can regulate cholesterol metabolism in ovarian cancer cells. Finally, a combination of time-resolved fluorescence resonance energy transfer, isothermal titration calorimetry, molecular docking, and molecular dynamics simulations were utilized to validate the ability of this peptide to bind to the epidermal growth factor receptor (EGFR) and impede the binding of epidermal growth factor (EGF) and EGFR.

Preferential binding of cucurbit[7]uril toward 2-amino-4-chlorophenol in chlorzoxazone

Chlorzoxazone (CLZ) is a centrally acting skeletal muscle relaxant synthesized from 2-amino-4-chlorophenol (ACP). However, due to the toxic side effects of ACP, its concentration in CLZ formulations is strictly regulated (0.5%). In this study, we investigate the preferential binding of cucurbit[7]uril (CB7) toward CLZ and ACP using 1H NMR, isothermal titration calorimetry (ITC) and molecular dynamics (MD) simulations, to explore ​possible future uses of CB7, either for purifying CLZ from its toxic impurity (ACP) or in sensing ACP in CLZ. ITC experiments confirm the formation of 1:1 complexes, with favorable enthalpic and entropic contributions and binding constants of 1.1 × 106 and 4.9 × 104M−1 for ACP and CLZ, respectively, while 1H NMR and MD simulations reveal the guests in the complexes are included within the cavity of CB7. Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA) results further confirmed the favorability of CB7 toward ACP. Additional MD simulations reveal that CB6 and CB8 preferentially bind CLZ and ACP, respectively, with CB6 forming inclusion and exclusion complexes with CLZ and ACP, respectively, while CB8 forms stable 1:1 and 1:2 inclusion complexes with each guest.

Entropy Drives Interpolymer Association in Water: Insights into Molecular Mechanisms.

Interpolymer association in aqueous solutions is essential for many industrial processes, new materials design, and the biochemistry of life. However, our understanding of the association mechanism is limited. Classical theories do not provide molecular details, creating a need for detailed mechanistic insights. This work consolidates previous literature with complementary isothermal titration calorimetry (ITC) measurements and molecular dynamics (MD) simulations to investigate molecular mechanisms to provide such insights. The large body of ITC data shows that intermolecular bonds, such as ionic or hydrogen bonds, cannot drive association. Instead, polymer association is entropy-driven due to the reorganization of water and ions. We propose a unifying entropy-driven association mechanism by generalizing previously suggested polyion association principles to include nonionic polymers, here termed polydipoles. In this mechanism, complementary charge densities of the polymers are the common denominators of association, for both polyions and polydipoles. The association of the polymers results mainly from two processes: charge exchange and amphiphilic association. MD simulations indicate that the amphiphilic assembly alone is enough for the initial association. Our proposed mechanism is a step toward a molecular understanding of the formation of complexes between synthetic and biological polymers under ambient or biological conditions.

Active sparse Bayesian committee machine potential for isothermal-isobaric molecular dynamics simulations.

Recent advancements in machine learning potentials (MLPs) have significantly impacted the fields of chemistry, physics, and biology by enabling large-scale first-principles simulations. Among different machine learning approaches, kernel-based MLPs distinguish themselves through their ability to handle small datasets, quantify uncertainties, and minimize over-fitting. Nevertheless, their extensive computational requirements present considerable challenges. To alleviate these, sparsification methods have been developed, aiming to reduce computational scaling without compromising accuracy. In the context of isothermal and isobaric ML molecular dynamics (MD) simulations, achieving precise pressure estimation is crucial for reproducing reliable system behavior under constant pressure. Despite progress, sparse kernel MLPs struggle with precise pressure prediction. Here, we introduce a virial kernel function that significantly enhances the pressure estimation accuracy of MLPs. Additionally, we propose the active sparse Bayesian committee machine (BCM) potential, an on-the-fly MLP architecture that aggregates local sparse Gaussian process regression (SGPR) MLPs. The sparse BCM potential overcomes the steep computational scaling with the kernel size, and a predefined restriction on the size of kernel allows for fast and efficient on-the-fly training. Our advancements facilitate accurate and computationally efficient machine learning-enhanced MD (MLMD) simulations across diverse systems, including ice-liquid coexisting phases, Li10Ge(PS6)2 lithium solid electrolyte, and high-pressure liquid boron nitride.

Insights into the molecular mechanism of dextran sulfate inhibiting DNA digestion by pepsin: a crucial role of sulfate group on the binding to DNA

Sulfate polysaccharides can inhibit DNA digestion in simulated gastric juice in vitro, which is important for regulating dietary nucleic acids metabolism, but the mechanism of inhibition is unclear. This study used dextran sulfate (DS) with different sulfate groups and molecular weights to explore the effect of DS on DNA digestion. Molecular interactions between DS and DNA were investigated by biolayer interferometry (BLI), isothermal titration calorimetry (ITC) and molecular dynamics simulations. Results indicated that DS with higher molecular weight and sulfate group content showed stronger inhibitory effect of DNA digestion. ITC results showed that the combined Kd value of DNA and DS was about 2.53 mM. The main reason for inhibition of DNA digestion is that the formation of hydrogen bonds between the sulfate group of DS and DNA bases hinders the binding of DNA to pepsin. This finding will facilitate new strategies for nucleic acid metabolism and oral drug delivery. Communicated by Ramaswamy H. Sarma

Gallic acid inhibits filament formation and promotes the disassembly of superoxide dismutase 1, a protein involved in the pathogenesis of amyotrophic lateral sclerosis

Superoxide dismutase 1 (SOD1) forms amyloid-like filaments in motor neurons, contributing to the pathogenesis of Amyotrophic Lateral Sclerosis (ALS) and leading to neuronal cell death. Earlier studies identified Epigallocatechin gallate (EGCG) as an effective inhibitor of SOD1 filaments. This study explored the potential of natural compounds to inhibit SOD1 filament formation as an ALS treatment strategy. Our investigation revealed that gallic acid, a soluble constituent of tannic acid and a hydrolysis product of EGCG, disrupts SOD1 filament formation, similar to EGCG. Furthermore, gallic acid demonstrated the ability to dismantle pre-existing SOD1 protein filaments. The study unveiled the inhibitory potential of red wine, a dietary source of tannic acid, and showed that tannic acid, when treated with tannase from lactobacilli, can enhance the inhibition against SOD1 filament formation. The findings were validated using isothermal titration calorimetry (ITC) and molecular dynamics (MD) simulations, suggesting that gallic acid can bind to both the soluble and filamentous forms of SOD1 protein. This interaction could elucidate the mechanism by which SOD1 filament formation is inhibited. In conclusion, the study unveils the molecular mechanism by which gallic acid inhibits the formation of SOD1 filaments. Our results suggest that gallic acid, a hydrolysis product of red wine, may be employed as a functional food ingredient to mitigate neurodegenerative diseases caused by protein filaments, such as ALS.

Discovery of anti-allergic components in Guomingkang Formula using sensitive HEMT biochips coupled with in vitro and in vivo validation

BackgroundAllergic rhinitis (AR) is a prevalent allergic disease, which seriously affects the sufferers’ life quality and increases the socioeconomic burden. Guominkang (GMK), a well-known prescription for AR treatment, showed satisfactory effects; while its anti-allergic components remain to be disclosed. AlGaN/GaN HEMT biochip is more sensitive and cost-effective than other binding equipments, indicating its great potential for screening of active ingredients from herbal medicines. MethodsAR mouse models were first established to test the anti-allergic effect of GMK and discover the ingredients absorbed into blood by ultra-high performance liquid chromatography-mass spectra (UHPLC-MS). Then, novel Syk/Lyn/Fyn-functionalized high electron mobility transistor (HEMT) biochips with high sensitivity and specificity were constructed and applied to screen the active components. Finally, the results from HEMT biochips screening were validated via in silico (molecular docking and molecular dynamics simulation), in vitro (RBL-2H3 cells), and in vivo (PCA mice model) assays. ResultsGMK showed a potent therapeutic effect on AR mice, and fifteen components were identified from the medicated plasma. Furthermore, hamaudol was firstly found to selectively inhibit the Syk and Lyn, and emodin was to selectively inhibit Lyn, which were further confirmed by isothermal titration calorimetry, molecular docking, and molecular dynamics simulation analyses. Suppression of the activation of FcεRI-MAPK signals might be the possible mechanism of the anti-allergic effect of hamaudol. ConclusionsThe targets of emodin and hamaudol were discovered by HEMT biochips for the first time. This study provided a novel and effective strategy to discover active components in a complex herbal formula by using AlGaN/GaN HEMT biochips.

Polypropylene fiber grafted calcium alginate with mesoporous silica for adsorption of Bisphenol A and Pb2+

Polypropylene grafted calcium alginate with mesoporous silica (PP-g-CaAlg@SiO2) for adsorbing Bisphenol A (BPA) and Pb2+ was prepared by calcium chloride (CaCl2) crosslinking and hydrochloric acid solution treatment. The PP-g-CaAlg@SiO2 was characterized by SEM, TEM, BET, XRD, FTIR and TG. PP-g-CaAlg@SiO2 exhibited excellent adsorption capacity for BPA and Pb2+, because the formation of reticulated nanorod structure increased its specific surface area. Subsequently, the adsorption behaviours of BPA and Pb2+, including adsorption isotherms and adsorption kinetics, were investigated. Afterward, isothermal titration calorimetry (ITC) and molecular dynamics (MD) simulation were performed to explore the adsorption mechanism. The results indicated that hydrogen bonding played the leading role in the adsorption of BPA, while the bonding of Pb2+ to carboxyl group binding sites was the focus of Pb2+ adsorption. In addition, the adsorption capacity of PP-g-CaAlg@SiO2 was stable over 10 cycles.

Supramolecular Study of the Interactions between Malvidin-3-O-Glucoside and Wine Phenolic Compounds: Influence on Color.

Supramolecular study of the interactions between the major wine anthocyanin, malvidin-3-O-glucoside (Mv3G) and different wine phenolic compounds (quercetin 3-O-β-glucopyranoside (QG), caffeic acid, (-)-epicatechin, (+)-catechin, and gallic acid) has been performed at two different molar ratios (1:1 and 1:2) in acidic medium where flavylium cation predominates (pH ≤ 2). Color variations have been evaluated by differential colorimetry using CIELAB color space. These studies have been complemented with isothermal titration calorimetry assays and molecular dynamics simulations. The color of Mv3G flavylium cation is modified by the interaction with QG toward more bluish and intense colors. Interaction constants between the anthocyanin and the different phenolic compounds were obtained, ranging from 9.72 × 108 M-1 for QG to 1.50 × 102 M-1 for catechin. Hydrophobic interactions and H-bonds are the main driving forces in the pigment/copigment aggregation, except for the interactions where caffeic acid is involved, in which hydrophobic interactions acquire greater preponderance.

Design, Synthesis, and Characterization of a Novel 2'-5'-Linked Amikacin-Binding Aptamer: An Experimental and MD Simulation Study.

To extend the approach of using RNA aptamers as transient protective groups for the synthesis of novel small-molecule drug derivatives from the existing aminoglycosides, we incorporated 2'-5' phosphodiester backbone modification in a structurally known neomycin RNA aptamer and studied the binding of a series of aminoglycosides using isothermal calorimetry (ITC) and molecular dynamics (MD) simulation. Experimental characterization of amikacin, a commercially available and widely used aminoglycoside for treating bacterial infections, shows that the aptamer A1 with a 2'-5' linkage between G15 and U16 exhibits a sevenfold increase in binding affinity with a lower binding energy compared to the native aptamer. Molecular dynamics (MD) simulation studies rationalize that this noncanonical linkage generates a narrower binding pocket by creating a superspiral RNA helical structure, which improves the ligand's fit in the binding pocket. These results provide new insights into applying 2'-5' linkages to diversify functional RNA aptamers as noncovalent protective groups in the synthesis of aminoglycoside derivatives, which can be further extended to other current drug molecules and complex natural compounds to make new pools of drug candidates more efficiently.

Chlorine-resistance, reswelling after drying and molecule/ions separation properties of carboxyl multi-walled carbon nanotubes/calcium alginate composite hydrogel membrane

In practice sodium hypochlorite (NaClO) is often used to clean biological contamination from membrane surfaces. It is worth thinking about how to balance the mechanical strength and chlorine resistance of calcium alginate (CaAlg) hydrogel membrane during cleaning. In this study, carboxyl multi-walled carbon nanotubes/calcium alginate (CMWCNT/CaAlg) composite hydrogel membrane with chlorine-resistance ability was prepared. Performance of CMWCNT/CaAlg membrane in various aspects, including morphology and chlorine resistance, were characterized. In addition, vacuum drying and reswelling method enhanced the mechanical strength of the membrane. Moreover, the interactions mechanism between CMWCNT and NaAlg were investigated by isothermal titration calorimetry (ITC) and molecular dynamics (MD) simulation. Subsequently, the factors influencing membrane separation efficiency towards dye molecule/salt ions were analyzed. It turns out that the composite membrane could achieve nearly 100% high efficiency in the rejection of dye molecules with large molecular weight (Mw > 700 Da), while the rejection of salt was below 8%. Our findings demonstrated CMWCNT/CaAlg composite membrane not only has excellent anti-fouling and chlorine resistance, but also has a very broad application prospect in the separation of dye and salt.

Encapsulation of phenolics in β-lactoglobulin: Stability, antioxidant activity, and inhibition of advanced glycation end products

β-Lactoglobulin (β-LG) is a whey protein with multiple binding sites for bioactive phenolics. Encapsulation of bioactive components in β-LG could enhance their activity and stability. In this study, vanillic acid (VA), ferulic acid (FA), and quercetin (QT) were encapsulated in β-LG. The resulting complex was studied by fluorescence spectroscopy, isothermal titration calorimetry, molecular docking, and molecular dynamics simulations. The β-LG–tri-ligand complex was successfully formed via an exothermic binding process when the ligands were added in the sequence of FA, QT, and VA, or QT, FA, and VA. The stabilities of the bioactive components in the complex were greatly improved by encapsulation. Molecular dynamics simulations confirmed that the binding of ligand molecules could increase the stability of the β-LG structure and made it more rigid/inflexible. The antioxidant activity and inhibition activity of the β-LG–tri-ligand complex against advanced glycation end products were higher than those of a mixture of FA, QT, and VA. The β-LG–tri-ligand complex shows great potential in functional food applications.

On interactions in binary mixtures used in solvent extraction: Insights from combined Isothermal Titration Calorimetry experiments and Molecular Dynamics simulations

Despite its importance, the description of liquid–liquid extraction is made difficult because the link between the experimental methods and the interactions present in the solution is not direct due to the many organisation phenomena that can occur. We propose a new methodology combining microcalorimetric measurements and molecular modelling to better describe the nature and strength of these interactions in these complex fluids by focusing on extractant-diluent mixtures (N,N-dialkylamides + n-alkanes). Enthalpies of dilution were obtained by microcalorimetric measurements at 298 K for each n-dodecane and n-heptane with DEHiBA and for n-dodecane with DEHBA. These results have been compared, thanks to a new method that we present here, to molecular dynamics simulations carried out on these same systems. We especially analyzed the influence of the alkyl branching of the extracting molecules and the role of the diluent. Globally, the excess enthalpy is such that these mixtures are energetically not very different from regular solutions but the detailed balance shows an important attraction between the extractants, which can lead to the formation of macromolecular entities favouring extraction. Calorimetry appears to be a method of choice for linking experiments and molecular modelling in these complex liquid mixtures, and experimental developments could benefit from using it more often, especially to validate force-fields.

Quantum polyamorphism in compressed distinguishable helium-4.

We demonstrate that two amorphous solid states can exist in 4He consisting of distinguishable Boltzmann atoms under compressed conditions. The isothermal compression of normal or supercritical fluid 4He was conducted at 3-25K using the isobaric-isothermal path integral centroid molecular dynamics simulation. The compression of fluid first produced the low-dispersion amorphous (LDA) state possessing modest extension of atomic necklaces. Further isothermal compression up to the order of 10 kbar to 1 Mbar or an isobaric cooling of LDA induced the transition to the high-dispersion amorphous (HDA) state. The HDA was characterized by long quantum wavelengths of atoms extended over several Angstroms and the promotion of atomic residual diffusion. They were related to the quantum tunneling of atoms bestriding the potential saddle points in this glass. The change in pressure or temperature induced the LDA-HDA transition reversibly with hysteresis, while it resembled the coil-globule transition of classical polymers. The HDA had lower kinetic and higher Gibbs free energies than the LDA at close temperature. The HDA was absent at T ≥ 13 K, while the LDA-HDA transition pressure significantly decreased with lowering temperature. The LDA and HDA correspond to the trapped and tunneling regimes proposed by Markland et al. [J. Chem. Phys. 136, 074511 (2012)], respectively. The same reentrant behavior as they found was observed for the expansion factor of the quantum wavelength as well as for atomic diffusivity.

Crystallisation competition between cubic and hexagonal ice structures: molecular-dynamics insight

ABSTRACT Under ambient pressure, two different ice polytypes of ice, cubic (Ic ) and hexagonal (Ih ) can be crystallised. Here, we elucidate the mechanism and kinetics of the phase transition of cubic ice with no stacking disorder to a perfect hexagonal ice at various temperatures, via molecular simulation. The study was performed through non-isothermal and isothermal molecular-dynamics simulations; these conditions were enforced on an equilibrated Ic /LDA/Ih system. Different results have been drawn from analysis in terms of local density, annealing kinetics, and phase transition. The evolution from cubic to hexagonal ice becomes prevalent at higher temperatures during the performed non-isothermal studies; to this end, a model-free kinetic study was proposed to address this phenomenon’s kinetic properties over a characteristic temperature range. Ice crystallisation is also studied in solid/liquid systems, where cubic or hexagonal ice was in contact with liquid water at sub-zero temperatures. It was revealed that water molecules have a propensity to follow and crystalise in the existing ice structure rather than forming a new interface.

Structural Basis of Inhibition of the Pioneer Transcription Factor NF-Y by Suramin.

NF-Y is a transcription factor (TF) comprising three subunits (NF-YA, NF-YB, NF-YC) that binds with high specificity to the CCAAT sequence, a widespread regulatory element in gene promoters of prosurvival, cell-cycle-promoting, and metabolic genes. Tumor cells undergo “metabolic rewiring” through overexpression of genes involved in such pathways, many of which are under NF-Y control. In addition, NF-YA appears to be overexpressed in many tumor types. Thus, limiting NF-Y activity may represent a desirable anti-cancer strategy, which is an ongoing field of research. With virtual-screening docking simulations on a library of pharmacologically active compounds, we identified suramin as a potential NF-Y inhibitor. We focused on suramin given its high water-solubility that is an important factor for in vitro testing, since NF-Y is sensitive to DMSO. By electrophoretic mobility shift assays (EMSA), isothermal titration calorimetry (ITC), STD NMR, X-ray crystallography, and molecular dynamics (MD) simulations, we showed that suramin binds to the histone fold domains (HFDs) of NF-Y, preventing DNA-binding. Our analyses, provide atomic-level detail on the interaction between suramin and NF-Y and reveal a region of the protein, nearby the suramin-binding site and poorly conserved in other HFD-containing TFs, that may represent a promising starting point for rational design of more specific and potent inhibitors with potential therapeutic applications.

Unsupervised Learning-Based Multiscale Model of Thermochemistry in 1,3,5-Trinitro-1,3,5-triazinane (RDX).

The response of high-energy-density materials to thermal or mechanical insults involves coupled thermal, mechanical, and chemical processes with disparate temporal and spatial scales that no single model can capture. Therefore, we developed a multiscale model for 1,3,5-trinitro-1,3,5-triazinane, RDX, where a continuum description is informed by reactive and nonreactive molecular dynamics (MD) simulations to describe chemical reactions and thermal transport. Reactive MD simulations under homogeneous isothermal and adiabatic conditions are used to develop a reduced-order chemical kinetics model. Coarse graining is done using unsupervised learning via non-negative matrix factorization. Importantly, the components resulting from the analysis can be interpreted as reactants, intermediates, and products, which allows us to write kinetics equations for their evolution. The kinetics parameters are obtained from isothermal MD simulations over a wide temperature range, 1200-3000 K, and the heat evolved is calibrated from adiabatic simulations. We validate the continuum model against MD simulations by comparing the evolution of a cylindrical hotspot 10 nm in diameter. We find excellent agreement in the time evolution of the hotspot temperature fields both in cases where quenching is observed and at higher temperatures for which the hotspot transitions into a deflagration wave. The validated continuum model is then used to assess the criticality of hotspots involving scales beyond the reach of atomistic simulations that are relevant to detonation initiation.