Differential Scanning Calorimetry (DSC) has been used in the past to study the thermal unfolding of many different viruses. Here we present the first DSC analysis of rabies virus. We show that non-inactivated, purified rabies virus unfolds cooperatively in two events centered at approximately 62 and 73 °C. Beta-propiolactone (BPL) treatment does not alter significantly viral unfolding behavior, indicating that viral inactivation does not alter protein structure significantly. The first unfolding event was absent in bromelain treated samples, causing an elimination of the G-protein ectodomain, suggesting that this event corresponds to G-protein unfolding. This hypothesis was confirmed by the observation that this first event was shifted to higher temperatures in the presence of three monoclonal, G-protein specific antibodies. We show that dithiothreitol treatment of the virus abolishes the first unfolding event, indicating that the reduction of G-protein disulfide bonds causes dramatic alterations to protein structure. Inactivated virus samples heated up to 70 °C also showed abolished recognition of conformational G-protein specific antibodies by Surface Plasmon Resonance analysis. The sharpness of unfolding transitions and the low standard deviations of the Tm values as derived from multiple analysis offers the possibility of using this analytical tool for efficient monitoring of the vaccine production process and lot to lot consistency.

Recently, there has been a noticeable increase in the applications of composite mixtures containing molten salt and solid oxide for thermal energy conversion and storage systems. This highlights that thermal conductivity of the composites are central for the purpose of designing and devising processes. Measuring the thermal conductivity of molten samples at elevated temperatures remains challenging. In this study, the possibility to use heat flux differential scanning calorimetry (DSC) to measure the thermal conductivity of molten samples at elevated temperatures is reported for the first time. The thermal conductivity of composite mixtures containing eutectic (Li,Na)2CO3 with and without selected solid oxides at ~675 °C was determined by using the proposed DSC approach. This mixture is a candidate for high temperature waste heat conversion to electric energy. In the DSC measurement program, steps with repeated thermal cycles between 410 and 515 °C were included to limit the effect of the interface thermal contact resistance. The determined values 0.826 ± 0.001, and 0.077 ± 0.004 W m-1K-1 for the carbonate mixtures with and without solid MgO were found to match the reliable analysis at similar conditions.

Thermal denaturation of Escherichia coli maltodextrin glucosidase was studied by differential scanning calorimetry, circular dichroism (230 nm), and UV-absorption measurements (340 nm), which were respectively used to monitor heat absorption, conformational unfolding, and the production of solution turbidity. The denaturation was irreversible, and the thermal transition recorded at scan rates of 0.5-1.5 K/min was significantly scan-rate dependent, indicating that the thermal denaturation was kinetically controlled. The absence of a protein-concentration effect on the thermal transition indicated that the denaturation was rate-limited by a mono-molecular process. From the analysis of the calorimetric thermograms, a one-step irreversible model well represented the thermal denaturation of the protein. The calorimetrically observed thermal transitions showed excellent coincidence with the turbidity transitions monitored by UV-absorption as well as with the unfolding transitions monitored by circular dichroism. The thermal denaturation of the protein was thus rate-limited by conformational unfolding, which was followed by a rapid irreversible formation of aggregates that produced the solution turbidity. It is thus important to note that the absence of the protein-concentration effect on the irreversible thermal denaturation does not necessarily means the absence of protein aggregation itself. The turbidity measurements together with differential scanning calorimetry in the irreversible thermal denaturation of the protein provided a very effective approach for understanding the mechanisms of the irreversible denaturation. The Arrhenius-equation parameters obtained from analysis of the thermal denaturation were compared with those of other proteins that have been reported to show the one-step irreversible thermal denaturation. Maltodextrin glucosidase had sufficiently high kinetic stability with a half-life of 68 days at a physiological temperature (37°C).

Starch is one of the main carbohydrates in food; it is formed by two polysaccharides: amylose and amylopectin. The granule size of starch varies with different botanical origins and ranges from less than 1 μm to more than 100 μm. Some physicochemical and functional properties vary with the size of the granule, which makes it of great interest to find an efficient and accurate size-based separation method. In this study, the full-feed depletion mode of split-flow thin cell fractionation (FFD-SF) was employed for a size-based fractionation of two types of starch granules (corn and potato) on a large scale. The fractionation efficiency (FE) of fraction-a for corn and potato granules was 98.4 and 99.4%, respectively. The FFD-SF fractions were analyzed using optical microscopy (OM) and gravitational field-flow fractionation (GrFFF). The respective size distribution results were in close agreement for the corn starch fractions, while they were slightly different for the potato starch fractions. The thermal properties of FFD-SF fractions were analyzed, and the results for the potato starch showed that the peak temperature of gelatinization (Tp) slightly decreases as the size of the granules increases. Additionally, the enthalpy of gelatinization (ΔH) increases when the granule size increases and shows negative correlation with the gelatinization range (ΔT).

DSC is used to determine thermally-induced conformational changes of biomolecules within a blood plasma sample. Recent research has indicated that DSC curves (or thermograms) may have different characteristics based on disease status and, thus, may be useful as a monitoring and diagnostic tool for some diseases. Since thermograms are curves measured over a range of temperature values, they are considered functional data. In this paper we apply functional data analysis techniques to analyze differential scanning calorimetry (DSC) data from individuals from the Lupus Family Registry and Repository (LFRR). The aim was to assess the effect of lupus disease status as well as additional covariates on the thermogram profiles, and use FD analysis methods to create models for classifying lupus vs. control patients on the basis of the thermogram curves.

Novel silicone-based nanocomposites with varied elastic properties were prepared by blending standard polydimethylsiloxane (PDMS) with a lower viscosity component (hydroxyl-terminated PDMS) and integrating a graphene nanoplatelets (GNP) filler modified by strands of deoxyribonucleic acid (DNA). The curing behavior of these nanocomposites was studied by dynamic and isothermal differential scanning calorimetry. The activation energies of the polymerization reactions were determined using the Kissinger method and two model-free isoconversional approaches, the Ozawa-Flynn-Wall and the Kissinger-Akahira-Sunose methods. Results show that the complex trend of the curing behavior can be described using the isoconversional methods, unveiling lower activation energies for the nanocomposites with standard PDMS matrices. The role of the DNA modification of graphene on the curing behavior is also demonstrated. The curing reactions of the nanocomposites with the PDMS matrix are favored by the presence of the GNP-DNA filler. PDMS/PDMS-OH blends generate softer nanocomposites with hardness and reduced elastic modulus that can be tuned by varying the amount of the filler.

Dioctadecyldimethylammonium bromide (DODAB) is a double chain cationic lipid, which assembles as bilayer structures in aqueous solution. The precise structures formed depend on, e.g., lipid concentration and temperature. We here combine differential scanning calorimetry (DSC) and X-ray scattering (SAXS and WAXS) to investigate the thermal and structural behavior of up to 120 mM DODAB in water within the temperature range 1-70 °C. Below 1 mM, this system is dominated by unilamellar vesicles (ULVs). Between 1 and 65 mM, ULVs and multilamellar structures (MLSs) co-exist, while above 65 mM, the MLSs are the preferred structure. Depending on temperature, DSC and X-ray data show that the vesicles can be either in the subgel (SG), gel, or liquid crystalline (LC) state, while the MLSs (with lattice distance d = 36.7 Å) consist of interdigitated lamellae in the SG state, and ULVs in the LC state (no Bragg peak). Critical temperatures related to the thermal transitions of these bilayer structures obtained in the heating and cooling modes are reported, together with the corresponding transition enthalpies.

The aim of this study was to conduct thermal characterization of sesame seeds and oils from various geographical origins (Ethiopia, India, Nigeria, Sudan, Turkey), different method of extraction (hexane and cold-pressing), and different types of derived products (halva and tahini). Thermal characterization was investigated using differential scanning calorimetry (DSC), which showed that origin of the seeds has no influence on the melting profile of sesame oil (peak temperature and enthalpy). Method of extraction (hexane and cold-pressing) influenced the peak temperatures of the resulting oils (p ≤ 0.05). The addition of 20% of palm olein to pure sesame oil influenced the significant changes in thermodynamic parameters such as peak temperature (Tm2), which was lowered from −5.89 °C to −4.99 °C, peak half width (T1/2), elevated from 3.01 °C to 4.52 °C, and the percentage of first peak area (% peak 1) lowered from 87.9 to 73.2% (p ≤ 0.05). The PCA method enabled to distinguish authentic and adulterated sesame oils of various origins. There were no significant differences in thermal properties among the products (halva, tahini) and the authentic sesame oil (p > 0.05). The obtained results showed DSC feasibility to characterize sesame oil and sesame products in terms of authenticity.

The purpose of the present paper was to assess the ability of five newly designed and synthesized meloxicam analogues to interact with phospholipid bilayers. Calorimetric and fluorescence spectroscopic measurements revealed that, depending on the details of the chemical structure, the studied compounds penetrated bilayers and affected mainly their polar/apolar regions, closer to the surface of the model membrane. The influence of meloxicam analogues on the thermotropic properties of DPPC bilayers was clearly visible because these compounds reduced the temperature and cooperativity of the main phospholipid phase transition. Additionally, the studied compounds quenched the fluorescence of prodan to a higher extent than laurdan, what pointed to a more pronounced interaction with membrane segments close to its surface. We presume that a more pronounced intercalation of the studied compounds into the phospholipid bilayer may be related to the presence of the molecule of a two-carbon aliphatic linker with a carbonyl group and fluorine substituent/trifluoromethyl group (compounds PR25 and PR49) or the three-carbon linker together with the trifluoromethyl group (PR50). Moreover, computational investigations of the ADMET properties have shown that the new meloxicam analogues are characterized by beneficial expected physicochemical parameters, so we may presume that they will have a good bioavailability after an oral administration.

Lab-scale investigations on the processing of small powder volumes are of special importance for applications in additive manufacturing (AM) techniques. Due to the technological importance of high-silicon electrical steel, and the increasing need for optimal near-net-shape AM processing, the aim of this study was to investigate the thermal behavior of a high-alloy Fe-Si powder for AM. An Fe-6.5wt%Si spherical powder was characterized using chemical, metallographic, and thermal analyses. Before thermal processing, the surface oxidation of the as-received powder particles was observed by metallography and confirmed by microanalysis (FE-SEM/EDS). The melting, as well as the solidification behavior of the powder, was evaluated using differential scanning calorimetry (DSC). Due to the remelting of the powder, a significant loss of silicon occurred. The morphology and microstructure analyses of the solidified Fe-6.5wt%Si revealed the formation of needle-shaped eutectics in a ferrite matrix. The presence of a high-temperature phase of silica was confirmed by the Scheil-Gulliver solidification model for the ternary model Fe-6.5wt%Si-1.0wt%O alloy. In contrast, for the binary model Fe-6.5wt%Si alloy, thermodynamic calculations predict the solidification exclusively with the precipitation of b.c.c. ferrite. The presence of high-temperature eutectics of silica in the microstructure is a significant weakness for the efficiency of the magnetization processes of soft magnetic materials from the Fe-Si alloy system.

Early detection of lung cancer (LC) significantly increases the likelihood of successful treatment and improves LC survival rates. Currently, screening (mainly low-dose CT scans) is recommended for individuals at high risk. However, the recent increase in the number of LC cases unrelated to the well-known risk factors, and the high false-positive rate of low-dose CT, indicate a need to develop new, non-invasive methods for LC detection. Therefore, we evaluated the use of differential scanning calorimetry (DSC) for LC patients' diagnosis and predicted survival. Additionally, by applying mass spectrometry, we investigated whether changes in O- and N-glycosylation of plasma proteins could be an underlying mechanism responsible for observed differences in DSC curves of LC and control subjects. Our results indicate selected DSC curve features could be useful for differentiation of LC patients from controls with some capable of distinction between subtypes and stages of LC. DSC curve features also correlate with LC patients' overall/progression free survival. Moreover, the development of classification models combining patients' DSC curves with selected plasma protein glycosylation levels that changed in the presence of LC could improve the sensitivity and specificity of the detection of LC. With further optimization and development of the classification method, DSC could provide an accurate, non-invasive, radiation-free strategy for LC screening and diagnosis.

Novel soluble liquid tin(ii) n-butoxide (Sn(OnC4H9)2), tin(ii) n-hexoxide (Sn(OnC6H13)2), and tin(ii) n-octoxide (Sn(OnC8H17)2) initiators were synthesized for use as coordination-insertion initiators in the bulk ring-opening polymerization (ROP) of l-lactide (LLA). In order to compare their efficiencies with the more commonly used tin(ii) 2-ethylhexanoate (stannous octoate, Sn(Oct)2) and conventional tin(ii) octoate/n-alcohol (SnOct2/nROH) initiating systems, kinetic parameters derived from monomer conversion data were obtained from non-isothermal differential scanning calorimetry (DSC). In this work, the three non-isothermal DSC kinetic approaches including dynamic (Kissinger, Flynn-Wall, and Ozawa); isoconversional (Friedman, Kissinger-Akahira-Sunose (KAS) and Ozawa-Flynn-Wall (OFW)); and Borchardt and Daniels (B/D) methods of data analysis were compared. The kinetic results showed that, under the same conditions, the rate of polymerization for the 7 initiators/initiating systems was in the order of liquid Sn(OnC4H9)2 > Sn(Oct)2/nC4H9OH > Sn(Oct)2 ≅ liquid Sn(OnC6H13)2 > Sn(Oct)2/nC6H13OH ≅ liquid Sn(OnC8H17)2 > Sn(Oct)2/nC8H17OH. The lowest activation energies (E a = 52, 59, and 56 kJ mol-1 for the Kissinger, Flynn-Wall, and Ozawa dynamic methods; E a = 53-60, 55-58, and 60-62 kJ mol-1 for the Friedman, KAS, and OFW isoconversional methods; and E a = 76-84 kJ mol-1 for the B/D) were found in the polymerizations using the novel liquid Sn(OnC4H9)2 as the initiator, thereby showing it to be the most efficient initiator in the ROP of l-lactide.

Differential scanning calorimetry (DSC) has emerged as a helpful technique both to characterize drug delivery systems and to study their interactions with bio-membranes. In this work, we compared idebenone (IDE)-loaded solid lipid nanoparticle (SLN) interactions with bio-membranes assessed by DSC with previous in vitro skin penetration data to evaluate the feasibility of predicting IDE skin penetration using DSC analyses. In vitro interactions experiments were performed using multi-lamellar liposomes as a model of bio-membrane. Enthalpy changes (ΔH) and transition temperature (Tm) were assessed during nine repeated DSC scans to evaluate IDE-loaded SLN⁻bio-membrane interactions over time. Analyzing ΔH and Tm values for each scan, we observed that the difference of ΔH and Tm values between the first and the last scan seemed to be related to SLN ability to locate IDE in the epidermis and in the stratum corneum, respectively. Therefore, the results of this study suggest the possibility of qualitatively predicting in vitro IDE skin penetration from IDE-loaded SLN utilizing the calorimetric parameters obtained from interaction experiments between the carriers under investigation and a model of bio-membrane.

In this work, we explore the ability to generate well-defined poly(butyl methacrylate) (PBMA) nanostructures by "in situ" polymerization of butyl methacrylate monomer (BMA). PBMA nanostructures of high and low aspect ratios have been successfully obtained through the free radical polymerization (FRP) of a BMA monomer in anodic aluminum oxide (AAO) nanoreactors of suitable size. A polymerization kinetics process has been followed by differential scanning calorimetry (DSC) and proton Nuclear Magnetic Resonance spectroscopy (1H-NMR).The determination of the kinetics of polymerization through DSC is based on a quick and direct analysis of the exothermic polymerization process, whereas the analysis through 1H-NMR also allows the unambiguous chemical analysis of the resulting polymer. When compared to bulk polymerization, both techniques demonstrate confinement effects. Moreover, DSC and 1H-NMR analysis give the same kinetics results and show a gel-effect in all the cases. The number average molecular weight (Mn) of the PBMA obtained in AAO of 60-300 nm are between 30·103-175·103 g/mol. Even if the Mn value is lower with respect to that obtained in bulk polymerization, it is high enough to maintain the polymer properties. As determined by SEM morphological characterization, once extracted from the AAO nanoreactor, the polymer nanostructures show controlled homogeneous aspect/size all throughout the length of nanopillar over a surface area of few cm2. The Young's modulus of low aspect ratio PBMA nanopillars determined by AFM gives a value of 3.1 ± 1.1 MPa. In this work, a 100% of PBMA polymer nanostructures are obtained from a BMA monomer in AAO templates through a quick double process: 30 min of monomer immersion at room temperature and 90 min of polymerization reaction at 60 °C. While the same nanostructures are obtained by polymer infiltration of PBMA at 200 °C in about 6 h, polymerization conditions are much softer than those corresponding to the polymer infiltration process. Furthermore, the 1H-NMR technique has been consolidated as a tool for studying the kinetics of the copolymerization reactions in confinement and the determination of monomer reactivity ratios.

Some of the best nucleating agents in nature are ice-nucleating proteins, which boost ice growth better than any other material. They can induce immersion freezing of supercooled water only a few degrees below 0 °C. An open question is whether this ability also extends to the deposition mode, i.e., to water vapor. In this work, we used three proteins, apoferritin, InaZ (ice nucleation active protein Z), and myoglobin, of which the first two are classified as ice-nucleating proteins for the immersion freezing mode. We studied the ice nucleation ability of these proteins by differential scanning calorimetry (immersion freezing) and by environmental scanning electron microscopy (deposition freezing). Our data show that InaZ crystallizes water directly from the vapor phase, while apoferritin first condenses water in the supercooled state, and subsequently crystallizes it, just as myoglobin, which is unable to nucleate ice.

Coproporphyrin ferrochelatases (CpfCs, EC 4.99.1.9) insert ferrous iron into coproporphyrin III yielding coproheme. CpfCs are utilized by prokaryotic, mainly monoderm (Gram-positive) bacteria within the recently detected coproporphyrin-dependent (CPD) heme biosynthesis pathway. Here, we present a comprehensive study on CpfC from Listeria monocytogenes (LmCpfC) including the first crystal structure of a coproheme-bound CpfC. Comparison of crystal structures of apo-LmCpfC and coproheme-LmCpfC allowed identification of structural rearrangements and of amino acids involved in tetrapyrrole macrocycle and Fe2+ binding. Differential scanning calorimetry of apo-, coproporphyrin III-, and coproheme-LmCpfC underline the pronounced noncovalent interaction of both coproporphyrin and coproheme with the protein (ΔTm  = 11 °C compared to apo-LmCpfC), which includes the propionates (p2, p4, p6, p7) and the amino acids Arg29, Arg45, Tyr46, Ser53, and Tyr124. Furthermore, the thermodynamics and kinetics of coproporphyrin III and coproheme binding to apo-LmCpfC is presented as well as the kinetics of insertion of ferrous iron into coproporphyrin III-LmCpfC that immediately leads to formation of ferric coproheme-LmCpfC (kcat /KM  = 4.7 × 105  m-1 ·s-1 ). We compare the crystal structure of coproheme-LmCpfC with available structures of CpfCs with artificial tetrapyrrole macrocycles and discuss our data on substrate binding, iron insertion and substrate release in the context of the CPD heme biosynthesis pathway. ENZYME: EC 4.99.1.9 DATABASE: pdb-codes of structural data in this work: 6RWV, 6SV3.

Calcium electroporation is a novel anti-cancer treatment investigated in clinical trials. We explored cell sensitivity to calcium electroporation and electroporation with bleomycin, using viability assays at different time and temperature points, as well as heat calorimetry, lipidomics, and flow cytometry. Three cell lines: HT29 (colon cancer), MDA-MB231 (breast cancer), and HDF-n (normal fibroblasts) were investigated for; (a) cell survival dependent on time of addition of drug relative to electroporation (1.2 kV/cm, 8 pulses, 99 µs, 1 Hz), at different temperatures (37 °C, 27 °C, 17 °C); (b) heat capacity profiles obtained by differential scanning calorimetry without added calcium; (c) lipid composition by mass spectrometry; (d) phosphatidylserine in the plasma membrane outer leaflet using flow cytometry. Temperature as well as time of drug administration affected treatment efficacy in HT29 and HDF-n cells, but not MDA-MB231 cells. Interestingly the HT29 cell line displayed a higher phase transition temperature (approximately 20 °C) versus 14 °C (HDF-n) and 15 °C (MDA-MB231). Furthermore the HT29 cell membranes had a higher ratio of ethers to esters, and a higher expression of phosphatidylserine in the outer leaflet. In conclusion, lipid composition and heat capacity of the membrane might influence permeabilisation of cells and thereby the effect of calcium electroporation and electrochemotherapy.

Intraoperative magnetic resonance imaging (MRI) has been proposed as a method to optimize intracerebral targeting and for tracking infusate distribution in gene therapy trials for nervous system disorders. We thus investigated possible effects of two MRI contrast agents, gadoteridol (Gd) and galbumin (Gab), on the distribution and levels of transgene expression in the rat striatum and their effect on integrity and stability of recombinant adeno-associated virus (rAAV) particles. MRI studies showed that contrast agent distribution did not predict rAAV distribution. However, green fluorescent protein (GFP) immunoreactivity revealed an increase in distribution of rAAV5-GFP, but not rAAV2-GFP, in the presence of Gd when compared with viral vector injected alone. In contrast, Gab increased the distribution of rAAV2-GFP not rAAV5-GFP. These observations pointed to a direct effect of infused contrast agent on the rAAV particles. Negative-stain electron microscopy (EM), DNAase treatment, and differential scanning calorimetry (DSC) were used to monitor rAAV2 and rAAV5 particle integrity and stability following contrast agent incubation. EMs of rAAV2-GFP and rAAV5-GFP particles pretreated with Gd appear morphologically similar to the untreated sample; however, Gab treatment resulted in surface morphology changes and aggregation. A compromise of particle integrity was suggested by sensitivity of the packaged genome to DNAase treatment following Gab incubation but not Gd for both vectors. However, neither agent significantly affected particle stability when analyzed by DSC. An increase in T m was observed for AAV2 in lactated Ringer's buffer. These results thus highlight potential interactions between MRI contrast agents and AAV that might affect vector distribution and stability, as well as the stabilizing effect of lactated Ringer's solution on AAV2.

The fabrication of nanocomposite films and fibers based on cellulose nanocrystals (P-tCNCs) and a thermoplastic polyurethane (PU) elastomer is reported. High-aspect-ratio P-tCNCs were isolated from tunicates using phosphoric acid hydrolysis, which is a process that affords nanocrystals displaying high thermal stability. Nanocomposites were produced by solvent casting (films) or melt-mixing in a twin-screw extruder and subsequent melt-spinning (fibers). The processing protocols were found to affect the orientation of both PU hard segments and the P-tCNCs within the PU matrix and therefore the mechanical properties. While the films were isotropic, both the polymer matrix and the P-tCNCs proved to be aligned along the fiber direction in the fibers, as shown using SAXS/WAXS, angle-dependent Raman spectroscopy, and birefringence analysis. Tensile tests reveal that fibers and films, at similar P-tCNC contents, display Young's moduli and strain-at-break that are within the same order of magnitude, but the stress-at-break was found to be ten-times higher for fibers, conferring them a superior toughness over films.

The purpose of this study is to assess water-polymer interaction in synthesized starch-derived superabsorbent polymer (S-SAP) for the treatment of solid waste sludge. While S-SAP for solid waste sludge treatment is still rare, it offers a lower cost for the safe disposal of sludge into the environment and recycling of treated solid as crop fertilizer. For that to be possible, the water-polymer interaction on S-SAP must first be fully comprehended. In this study, the S-SAP was prepared through graft polymerization of poly (methacrylic acid-co-sodium methacrylate) on the starch backbone. By analyzing the amylose unit, it was possible to avoid the complexity of polymer networks when considering S-SAP using molecular dynamics (MD) simulations and density functional theory (DFT). Through the simulations, formation of hydrogen bonding between starch and water on the H06 of amylose was assessed for its flexibility and less steric hindrance. Meanwhile, water penetration into S-SAP was recorded by the specific radial distribution function (RDF) of atom-molecule interaction in the amylose. The experimental evaluation of S-SAP correlated with high water capacity by measuring up to 500% of distilled water within 80 min and more than 195% of the water from solid waste sludge for 7 days. In addition, the S-SAP swelling showed a notable performance of a 77 g/g swelling ratio within 160 min, while a water retention test showed that S-SAP was capable of retaining more than 50% of the absorbed water within 5 h of heating at 60 °C. The water retention of S-SAP adheres to pseudo-second-order kinetics for chemisorption reactions. Therefore, the prepared S-SAP might have potential applications as a natural superabsorbent, especially for the development of sludge water removal technology.