Peak Mass Research Articles

Role of CT-derived global longitudinal strain in identifying left ventricular dysfunction in patients with severe aortic stenosis and preserved left ventricular ejection fraction

Abstract Introduction In patients with severe aortic stenosis, LVEF <50% is a current class 1 indication for aortic valve replacement. There are however studies showing increased mortality in patients with asymptomatic severe aortic stenosis and LVEF between 50-59% [1, 2]. Global longitudinal strain (GLS) has emerged as a more sensitive predictor of systolic dysfunction than ejection fraction in a wide number of cardiac diseases [3]. Purpose The aim of our study was to analyze left ventricular global longitudinal strain (GLS) by cardiac computed tomography (CT) in patients with severe aortic stenosis. We hypothesized that a larger portion of patients with LVEF 50-59% have signs of systolic dysfunction by GLS as compared to patients with LVEF ≥60%. Methods We identified 238 patients (median age 81, 51.3% females) with severe aortic stenosis, assessed by cardiac CT before undergoing transcatheter aortic valve replacement at Uppsala University hospital between 2016-2019. Patients in atrial fibrillation was excluded due to suboptimal image quality. Left ventricular mass index (LVMi), EF and peak GLS were calculated. The patients were divided into three groups based on LVEF (Reduced LVEF <50%, Low-Normal LVEF 50-59% and Normal LVEF ≥60%) and compared with each other regarding GLS, biomarkers and background variables. The cut off for significantly reduced GLS was set at <16%, in accordance with recently proposed criteria [4]. Results As expected GLS was significantly reduced in all patients with LVEF <50%. Among patients with Low-Normal LVEF, 62.3% had significantly reduced GLS as compared to only 3.8% patients with Normal LVEF >60% (p < 0.001). Median GLS in Low-Normal LVEF was 15.5% (IQR 14.1 - 17.4) as compared to 20.2% (IQR 18.4 - 21.9) in Normal LVEF (p < 0.001). Patients with Low-Normal LVEF had a significantly higher LV mass as compared to patients with Normal LVEF (median LVMi 85.8 g/m² vs. 75.8 g/m², p < 0.001). NT-pro-BNP was significantly higher in patients with Low-Normal LVEF compared to Normal LVEF (median 1130 vs. 664 ng/l, p = 0.003). Conclusions Almost two third of patients with severe aortic stenosis and lower range of normal LVEF (50-59%) had abnormally reduced GLS, reflecting systolic LV dysfunction, which was uncommon among those with LVEF >60%. Patients with LVEF 50-59% also presented with higher LV mass index and higher levels of NT-pro-BNP than patients with normal LVEF >60%, reflecting hypertrophic response of the myocardium to pressure overload which over time may lead to progressive myocardial decompensation. LVEF in the range of 50-59% might be an early sign of LV decompensation in severe aortic stenosis.

Clean and Complete Protein Digestion with an Autolysis Resistant Trypsin for Peptide Mapping.

Peptide mapping requires cleavage of proteins in a predictable fashion so that target protein-specific peptides can be reliably identified and quantified. Trypsin, a commonly used protease in this process, can also undergo self-cleavage or autolysis, thereby reducing the effectivity and even cleavage specificity at lysine and arginine residues. Here, we report highly efficient and reproducible peptide mapping of biotherapeutic monoclonal antibodies. We highlight the properties of a homogeneous chemically modified trypsin on thermal stability, a 54% increase in melting temperature with an 84% increase in energy required for unfolding, an indication of more thermally stable trypsin, >90% retained intact mass peak area after exposure to digestion conditions confirming autolysis resistance, 10× more intensity for intact enzyme compared to trypsin of similar source and narrower molecular weight distribution with LC-MS indicative of low degradation compared to 3 other types of trypsin. Finally, we show the utility of this autolysis-resistant trypsin in characterizing biotherapeutic monoclonal antibodies consistently and reliably showing a >30% reduction in missed cleavage for a short-duration protein digestion time of 30 min compared to heterogeneously modified trypsin of a similar source.

Subhalos in Galaxy Clusters: Coherent Accretion and Internal Orbits

Subhalo dynamics in galaxy cluster host halos govern the observed distribution and properties of cluster member galaxies. We use the IllustrisTNG simulation to investigate the accretion and orbits of subhalos found in cluster-size halos. We find that the median change in the major axis direction of cluster-size host halos is approximately 80° between a ∼ 0.1 and the present day. We identify coherent regions in the angular distribution of subhalo accretion, and ∼68% of accreted subhalos enter their host halo through ∼38% of the surface area at the virial radius. The majority of galaxy clusters in the sample have ∼2 such coherent regions. We further measure angular orbits of subhalos with respect to the host major axis and use a clustering algorithm to identify distinct orbit modes with varying oscillation timescales. The orbit modes correlate with subhalo accretion conditions. Subhalos in orbit modes with shorter oscillations tend to have lower peak masses and accretion directions somewhat more aligned with the major axis. One orbit mode, exhibiting the least oscillatory behavior, largely consists of subhalos that accrete near the plane perpendicular to the host halo major axis. Our findings are consistent with expectations from inflow from major filament structures and internal dynamical friction: most subhalos accrete through coherent regions, and more massive subhalos experience fewer orbits after accretion. Our work offers a unique quantification of subhalo dynamics that can be connected to how the intracluster medium strips and quenches cluster galaxies.

Halogen Tailoring of Platinum Electrocatalyst with High CO Tolerance for Methanol Oxidation Reaction.

The catalytic activity of platinum for CO oxidation depends on the interaction of electron donation and back-donation at the platinum center. Here we demonstrate that the platinum bromine nanoparticles with electron-rich properties on bromine bonded with sp-C in graphdiyne (PtBr NPs/Br-GDY), which is formed by bromine ligand and constitutes an electrocatalyst with a high CO-resistant for methanol oxidation reaction (MOR). The catalyst showed peak mass activity for MOR as high as 10.4 A mgPt -1, which is 20.8 times higher than the 20 % Pt/C. The catalyst also showed robust long-term stability with slight current density decay after 100 hours at 35 mA cm-2. Structural characterization, experimental, and theoretical studies show that the electron donation from bromine makes the surface of platinum catalysts highly electron-rich, and can strengthen the adsorption of CO as well as enhance π back-donation of Pt to weaken the C-O bond to facilitate CO electrooxidation and enhance catalytic performance during MOR. The results highlight the importance of electron-rich structure among active sites in Pt-halogen catalysts and provide detailed insights into the new mechanism of CO electrooxidation to overcome CO poisoning at the Pt center on an orbital level.

Low-temperature treatment optimization for diesel-contaminated kaolin: Mutual impacts of generated pyrolytic carbon and particle agglomeration

Efficiency improvement in low-temperature treatment for diesel-contaminated sites is urgent because changes in soil properties and the generation of new substance during the remediation process can influence the duration and energy utilization. This paper focuses on low-temperature treatment optimization based on the mutual impacts of pyrolytic carbon and kaolin aggregation. Results reveal that the peak mass loss rate occurred between 100–150°C, with minimal loss beyond 200°C. Samples thermally treated at 150–200°C exhibited darker colors, indicating pyrolytic carbon formation, corroborated by x-ray photoelectron spectroscopy (XPS), fourier transform infrared spectroscopy (FTIR), and three-dimensional fluorescence spectrum (3D-EEM) analyses. Additionally, diesel contamination influenced the fractal dimension of aggregates by influencing adhesion forces (<10000 mg/kg) and forming liquid bridges (≥10000 mg/kg) in untreated kaolin, resulting in an initial increase and subsequent fall in fractal dimension with increasing concentration. Decline rates of pollutant gas concentration were closely correlated with fractal dimension changes under thermal conditions due to pollutant volatilization and pyrolytic carbon formation. Based on the consistency between fractal dimension and decline rate, two critical remediation concentrations (C0) and temperatures (T0) indices were identified to optimize the low-temperature remediation.

Brain gene expression reveals pathways underlying nocturnal migratory restlessness

Migration is one of the most extreme and energy demanding life history strategies to have evolved in the animal kingdom. In birds, champions of long-distance migrations, the seasonal emergence of the migratory phenotype is characterised by rapid physiological and metabolic remodelling, including substantial accumulation of fat stores and increases in nocturnality. The molecular underpinnings and brain adaptations to seasonal migrations remain poorly understood. Here, we exposed Common quails (Coturnix coturnix) to controlled changes in day length to simulate southward autumn migration, and then blocked the photoperiod until birds entered the non-migratory wintering phase. We first performed de novo RNA-Sequencing from selected brain samples (hypothalamus) collected from birds at a standardised time at night, either in a migratory state (when restlessness was highest and at their body mass peak), or in a non-migratory state and conducted differential gene expression and functional pathways analyses. We found that the migratory state was associated with up-regulation of a few, yet functionally well defined, gene expression networks implicated in fat trafficking, protein and carbohydrate metabolism. Further analyses that focused on candidate genes (apolipoprotein H or APOH, lysosomal associated membrane protein-2 or LAMP2) from samples collected during the day or night across the entire study population suggested differences in the expression of these genes depending on the time of the day with the largest expression levels being found in the migratory birds sampled at night. We also found that expression of APOH was positively associated with levels of nocturnal activity in the migratory birds; such an association was absent within the non-migratory birds. Our results provide novel experimental evidence revealing that hypothalamic changes in expression of apolipoprotein pathways, which regulate the circulatory transport of lipids, are likely key regulatory activators of nocturnal migratory movements. Our study paves the way for performing deeper functional investigations on seasonal molecular remodelling underlying the development of the migratory phenotype.

An Expedited Qualitative Profiling of Free Amino Acids in Plant Tissues Using Liquid Chromatography-Mass Spectrometry (LC-MS) in Conjunction With MS-DIAL.

The estimation of relative levels of amino acids is crucial for understanding various biological processes in plants, including photosynthesis, stress tolerance, and the uptake and translocation of nutrients. A wide range of liquid chromatography (LC; HPLC/UHPLC)-based methods is available for measuring the quantity of amino acids in plants. Additionally, the coupling of LC with mass spectrometry (MS) significantly enhanced the robustness of existing chromatographic methods used for amino acid quantification. However, accurate annotation and integration of mass peaks can be challenging for plant biologists with limited experience in analyzing MS data, especially in studies involving large datasets with multiple treatments and/or replicates. Further, there are instances when the experiment demands an overall view of the amino acids profile rather than focusing on absolute quantification. The present protocol provides a detailed LC-MS method for obtaining a qualitative amino acids profile using MS-DIAL, a versatile and user-friendly program for processing MS data. Free amino acids were extracted from the leaves of control and Tomato leaf curl Palampur virus (ToLCPalV)-infected Nicotiana benthamiana plants. Extracted amino acids were derivatized and separated using UHPLC-QTOF, with each amino acid subsequently identified by aligning mass data with a custom text library created in MS-DIAL. Further, MS-DIAL was employed for internal standard-based normalization to obtain a qualitative profile of 15 amino acids in control and virus-infected plants. The outlined method aims to simplify the processing of MS data to quickly assess any modulation in amino acid levels in plants with a higher degree of confidence.

Crystallographic Dependence of Field Evaporation Energy Barrier in Metals Using Field Evaporation Energy Loss Spectroscopy Mapping.

Atom probe tomography data are composed of a list of coordinates of the reconstructed atoms in the probed volume. The elemental identity of each atom is derived from time-of-flight mass spectrometry, with no local chemical information readily available. In this study, we use a data processing technique referred to as field evaporation energy loss spectroscopy (FEELS), which analyzes the tails of mass peaks. FEELS was used to extract critical energetic parameters that are related to the activation energy for atoms to escape from the surface under intense electrostatic field and dependent of the path followed by the departing atoms. We focused our study on pure face-centered cubic metals. We demonstrate that the energetic parameters can be mapped in two-dimensional with nanometric resolution. A dependence on the considered crystallographic planes is observed, with sets of planes of low Miller indices showing a lower sensitivity to the field. The temperature is also an important parameter in particular for aluminum, which we attribute to an energetic transition between two paths of field evaporation between 25 and 60 K close to (002) pole. This paper shows that the information that can be retrieved from the measured energy loss of surface atoms is important both experimentally and theoretically.

Current concept of osteoporosis diagnosis and treatment in children

Osteoporosis (OP) is a disease characterized by a decrease in bone strength, leading to an increased risk of fractures. Bone fragility in children may be due to genetic disorders, chronic underlying conditions or taking medications that negatively affect bone metabolism. The article considers the modern classification of OP in children, approaches to its diagnosis, the role of dual-energy X-ray absorptiometry for diagnosis.The main goal of OP prevention and treatment is to achieve a higher peak of bone mass, improve bone microarchitectonics, and, as in adults, reduce the risk of fractures, prevent skeletal deformities, improve mobility, independence and quality of life. Recommendations for lifestyle changes are presented, including adequate calcium intake and vitamin D, taking antiresorptive drugs, as well as prospective management of children with OP.

Mass and Mobility of Ions Produced by Radioactive Sources and Corona Discharges.

Positive and negative ions produced by radioactive sources and corona discharges in gases find a number of applications, including charging aerosol particles prior to their measurement by electrical and/or electrical mobility techniques. The degree to which these ions can charge aerosol particles depends on their mobility and mass; properties that are strongly affected by the composition of the carrier gas and the impurities that it contains. We show that when the purity of the carrier gas is increased, the mobility of both positive and negative ions increases by more than 50%, whereas the respective masses reduce by more than 50%. In most cases, the dominant positive species is N4+, whereas NO2- and NO3- prevail for the negative polarity. Differences in ion mobility and mass resulting from the two ionization methods (i.e., radioactive source and corona discharges) remain limited. When volatile methyl siloxanes (VMS) are introduced deliberately to the gas, the mobility of the cations decreases by 39% and their mass increases by 385%, while the dominant mobility and mass peaks of the negative ions remains almost unaffected. Interestingly, introduction of VMS also leads to consistent and reproducible positive ion properties across all variations of the experiments, which can be especially relevant for charging aerosol particles in a reproducible manner. Taken together, the new measurements we report in this paper corroborate prior knowledge that the composition and purity of the carrier gas strongly influence the properties of positive and negative ions generated in aerosol neutralizers, and provide new evidence regarding their evolution in the presence of impurities.

Supermassive Primordial Black Holes for Nano-Hertz Gravitational Waves and High-redshift JWST Galaxies

Recently, observational hints for supermassive black holes have been accumulating, prompting the question: Can primordial black holes (PBHs) be supermassive, particularly with masses M ≳ 109 M ⊙? A supercritical bubble, containing an inflating baby universe, that nucleated during inflation can evolve into a PBH in our observable universe. We find that when the inflaton slowly transitions past a neighboring vacuum, the nucleation rate of supercritical bubbles inevitably peaks, leading to a mass distribution of multiverse PBHs with a peak mass up to M ≳ 1011 M ⊙. Thus, our mechanism naturally provides a primordial origin for supermassive black holes.

Physicochemical Characterization of Religious Burning Aerosols in Lhasa on the Qinghai‐Tibet Plateau

AbstractReligious burning (RB) has been identified as a major source of atmospheric aerosols on the Qinghai‐Tibet Plateau. However, there is limited understanding of the detailed chemical composition, size distribution, and optical properties of RB aerosols in this region. To characterize these important aerosol properties, ambient PM2.5 and size resolved aerosols from RB emissions in Lhasa were collected during summer 2019. Organic functional group (OFG) and inorganic ion composition was measured using Fourier transform infrared spectroscopy and ion chromatography, respectively. The ambient PM2.5 was dominated by organic components, with the OFG concentrations significantly higher during religious events, reflecting the substantial impact of RB emissions on local air quality. The RB aerosols were characterized by high fractions of alkane (34%), hydroxyl (29%), and carboxylic acid (13%) groups, with peak mass in the accumulation mode (0.56–1.00 μm). The high abundance of hydroxyl group and the size distribution pattern suggested that the RB aerosols were formed from volatilization of fuel materials followed by unaltered condensation, a process that may be unique to the low‐temperature, low‐oxygen burning in the scattered burners at the temples. The absorption coefficient of RB aerosols showed similar size distribution to the mass size distribution, but the absorption Ångström exponent displayed the lowest value in the 0.56–1.00 μm size mode. This specific size distribution aligned with the mass fraction of carboxylic acids and mirrored the mass proportion of alkanes, suggesting that smaller and larger particles were enriched with substances that have higher light‐absorbing capabilities.

Classification of Latilactobacillus sakei subspecies based on MALDI-TOF MS protein profiles using machine learning models.

Latilactobacillus sakei plays a significant role in the realm of food bacteria. One particular subspecies of L. sakei is employed as a protective agent during food fermentation, whereas another strain is responsible for food spoilage. Hence, it is crucial to precisely differentiate between the two subspecies of L. sakei. In this study, machine learning models based on protein mass peaks were developed for the first time to distinguish L. sakei subspecies. Furthermore, the efficacy of three commonly used machine learning algorithms for microbial classification was evaluated. Our results provide the foundation for future research on developing machine learning models for the classification of microbial species or subspecies. In addition, the developed model can be used in the food industry to monitor L. sakei subspecies in fermented foods in a time- and cost-effective method for food quality and safety.

Experimental and numerical investigation on the combustion behavior of densified wood: Differences among wood species and impact of char oxidation

Densified wood (DW) is a novel structural material with promissing application potential in construction industry due to its excellent mechanical performance. However, it could also be a disastrous fuel contributing to the development of fires. This work experimentally and numerically investigated the combustion behavior of DW. The differences among wood species (fir, pine and oak) and their char oxidation impacts are emphasized. The experimental results indicate that DW has a 44.7 K lower pyrolysis peak temperature on average, a 32.2 % lower peak mass loss rate, and a 8 % higher residual mass ratio than natural wood (NW) in N2 due to the delignification process. The hemicellulose and cellulose pyrolysis are endothermic in N2 but exothermic in air. Wood pyrolysis has an additional char oxidation reaction in air. The higher density of DW leads to longer times for ignition and flaming combustion. DW presents a higher heat release rate and average 4.7 MJ kg−1 higher effective heat of combustion of char. Two parallel pyrolysis reaction schemes based on ambiance are proposed, which well-predict the microscale characterizations but fail to represent the combustion behavior. An optimized mode was developed by adding a continuous char oxidation reaction to the N2 reaction scheme, which best predicts the combustion behaviors.

Aqueous Synthesis of Au10Pt1 Nanorods Decorated with MnO2 Nanosheets for the Enhanced Electrocatalytic Oxidation of Methanol.

Developing novel catalysts with high activity and high stability for the methanol oxidation reaction (MOR) is of great importance for the ever-broader applications of methanol fuel cells. Herein, we present a facile technique for synthesizing Au10Pt1@MnO2 catalysts using a wet chemical method and investigate their catalytic performance for the MOR. Notably, the Au10Pt1@MnO2-M composite demonstrated a significantly high peak mass activity of 15.52 A mg(Pt)-1, which is 35.3, 57.5, and 21.9 times greater than those of the Pt/C (0.44 A mg(Pt)-1), Pd/C (0.27 A mg(Pt)-1), and Au10Pt1 (0.71 A mg(Pt)-1) catalysts, respectively. Comparative analysis with commercial Pt/C and Pd/C catalysts, as well as Au10Pt1 HSNRs, revealed that the Au10Pt1@MnO2-M composite exhibited the lowest initial potential, the highest peak current density, and superior CO anti-poisoning capability. The results demonstrate that the introduction of MnO2 nanosheets, with excellent oxidation capability, not only significantly increases the reactive sites, but also promotes the reaction kinetics of the catalyst. Furthermore, the high surface area of the MnO2 nanosheets facilitates charge transfer and induces modifications in the electronic structure of the composite. This research provides a straightforward and effective strategy for the design of efficient electrocatalytic nanostructures for MOR applications.

Simultaneous determination of sixteen phthalic acid esters (PAEs) in soil and evaluation of matrix effect using a QuEChERS/GC/MS-internal standard method.

Phthalic acid esters (PAEs) are emerging pollutants that need to be analyzed precisely. Chromatography-based determination of PAE content in soils are frequently affected by matrix effect, which may limit the quantification of different kinds of PAEs from different types of soil. Here we optimized a QuEChERS protocol combined with gas chromatography-mass spectrometry (GC-MS) for simultaneous determination of 16 PAEs in different soils. PAEs in different type of soils (fluvo-aquic soil, red soil, and black soil) were extracted with acetonitrile followed by GC-MS detection based on quantitative ion internal standard method. All 16 PAEs showed excellent linear relationships with mass peak areas (R2 > 0.99). The limits of detection (LOD) and limits of quantitation (LOQ) of all the samples were in the range of 0.91-66.97µg/kg and 2.7-200.9µg/kg, respectively. The accurate test at 0.5, 0.1, and 1.0mg/kg spiking level recorded recovery rate between 80.11% and 100.99% with relative standard deviations (RSDs) ranging from 0.37 to 8.50% in tested matrices. No significant matrix effect was observed for most tested PAEs. This is a simple method with high sensitivity and strong stability, which is suitable and reproducible for quantifying large number of PAEs in different types of soil.

Platinum-Ruthenium Bimetallic Nanoparticle Catalysts Synthesized Via Direct Joule Heating for Methanol Fuel Cells.

Platinum-Ruthenium (PtRu) bimetallic nanoparticles are promising catalysts for methanol oxidation reaction (MOR) required by direct methanol fuel cells. However, existing catalyst synthesis methods have difficulty controlling their composition and structures. Here, a direct Joule heating method to yield highly active and stable PtRu catalysts for MOR is shown. The optimized Joule heating condition at 1000°C over 50 microseconds produces uniform PtRu nanoparticles (6.32wt.% Pt and 2.97 wt% Ru) with an average size of 2.0 ± 0.5 nanometers supported on carbon black substrates. They have a large electrochemically active surface area (ECSA) of 239 m2 g-1 and a high ECSA normalized specific activity of 0.295mA cm-2. They demonstrate a peak mass activity of 705.9mA mgPt -1 for MOR, 2.8 times that of commercial 20 wt.% platinum/carbon catalysts, and much superior to PtRu catalysts obtained by standard hydrothermal synthesis. Theoretical calculation results indicate that the superior catalytic activity can be attributed to modified Pt sites in PtRu nanoparticles, enabling strong methanol adsorption and weak carbon monoxide binding. Further, the PtRu catalyst demonstrates excellent stability in two-electrode methanol fuel cell tests with 85.3% current density retention and minimum Pt surface oxidation after 24 h.

Numerical Investigation of the Effect of Intake Closing Timing on Flow Field and Combustion Process in an Elliptical Rotary Engine

Abstract The aim of this research is to investigate the effect of intake closing timing (ICT) on the flow field and combustion process in elliptical rotary engines. The model that can accurately describe the working process of the elliptical rotary engine was established, five kinds of ICTs were designed, and the influence of ICT on the flow field and combustion process was studied. The results show that the advance of the ICT can increase the intake mass flowrate and reduce the back flowrate, the volumetric efficiency is 86.1% at a 145-deg crank angle (°CA) before top dead center (BTDC), which is 7.6% higher than 125 °CA BTDC. The advance of the ICT improves the consumption speed, makes the combustion reaction more intense, and shortens the combustion time. When the ICT is 145 °CA BTDC, the crank angle when the burned mass fraction is 90% (CA90) is 19.4 °CA earlier than 125 °CA BTDC, the peak mass of hydroxy in a cylinder is 41.6% higher, and the peak pressure in a cylinder is 25.9% higher. With the advance of the ICT, the pressure and heat release in the cylinder are significantly increased, the peak temperature in the cylinder is increased, the rate of carbon monoxide generation is accelerated, and the mass of nitrogen oxide emission is significantly increased. However, advancing the ICT cannot improve the indicated thermal efficiency of the elliptical rotary engine. This analysis provides a comprehensive understanding of the ICT of elliptical rotary engines.

On the Synthesis, Characterization and Applications of Para-nitro Aniline-based Reactive Dye

The dye was synthesized by nitration of aniline to para-nitro aniline, diazotization and coupling reactions. The melting point of the para-nitro aniline was taken to yield 147ºC. The result from the GC-MS shows that the compound with the highest relative abundance of 100% showed a molecular ion peak of 129.4 compounds which correspond to lower relative abundance. Also from the GC-MS result, the two peaks of masses 57.5 g and 83.4 g respectively correspond to fragments of the samples. Ultra-violet spectrophotometer was used to determine the absorption of the dye. The para-nitro aniline dye as synthesized showed the highest absorbance of 2.214 at 220 nanometers. The dye was applied to pure cotton and cellulose. The percentage shift of the dye toward the fabric i.e. percentage exhaustion of the dye was calculated to yield 73% which shows that the reactive dye has high exhaustion value. Fastness test was carried out on the dyed fabric to characterize the material’s color resistance to fading and running. These tests are heat, light and wash fastness which is rated 1-5 according to Ukponwam and Odilora (2000) where 1 shows poor fastness and 5 very good fastness. The wash, heat and light fastness of 4, 5 and 5 respectively shows that the dyed fabric has an excellent fastness to washing, heating and sun drying.

Molecular analysis of recombinant collagenase from Bacillus siamensis strain Z1: Gene Cloning, expression and in-silico characterization

This study focuses on gene cloning, expression, biochemical and analytical characterization along with structural and functional characterization of collagenase followed with molecular docking, dynamics study and Molecular Mechanics Poisson-Boltzmann Surface Area (MMPBSA). The collagenase gene identified from the genome of the novel collagenase Bacillus siamensis strain Z1 is introduced into E.coli DH5α, subsequently expressed in E.coli BL21 (DE3) withisopropyl β −d – 1 − thiogalactopyranoside(IPTG) induction and further affinity purified yielding in ∼ 89.4 kDa recombinant collagenase which demonstrated alkali characteristics and thermostability determined by thermodynamic parameters. Recombinant collagenase revealed good stability when exposed to diverse biochemical components. The recombinant collagenase identity was confirmed through matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) showing specific mass peaks and via N-terminal sequence analysis as MTAVNQTISK. Moreover, the concluded N-terminal amino acid sequence from Edman degradation displayed significant resemblance. The structural and functional analysisof recombinant collagenase was analysed by Circular dichroism (CD), Proton nuclear magnetic resonance (1H NMR) spectrometry and Thermogravimetric analysis (TGA). The recombinant collagenase also showed gelatin liquefaction ability. The collagenase gene sequence is also assessed for structural and functional characterizations by using various computational tools and revealed its classification in U32 family peptidase. A Grand average of hydropathicity index (GRAVY) score of −0.295 and instability index of 37.22 was obtained. Homology model of collagenase gene was generated by employing SWISS-MODEL and structure analysis by Ramachandran plot. Molecular docking of modelled collagenase with four different substrates was carried out by PyRx and Autodocking. Highest docking score of −12.7 kcal/mol was obtained for Alaska pollock hydroxyproline containing marine collagen peptide (APHCP). Subsequently, Molecular dynamics and simulations for highest score docked complex was assessed using GROningenMAchine for Chemical Simulations (GROMACS).