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The study evaluated the application of a novel high-pressure microbial inactivation method combining dense carbon dioxide with modified atmosphere packaging on organic fresh-cut squash (Cucurbita moschata). Approximately 4 g or 32 g of squash was packed in plastic pouches filled with CO 2 to test two different gas-to-product ratios and treated with the high-pressure method at previously optimized process conditions (45 °C, 6.0 MPa and 40 min). The products were then stored for 21 days at 4 °C and assessed for enzymatic activity, product quality, sugar content, bioaccessibility (polyphenols, DPPH antioxidant activity, and carotenoids), and sensory acceptance, with products packed in air and CO 2 serving as controls. The high-pressure treatment effectively inactivated inoculated E. coli to undetectable levels (inactivation >3.63 ± 0.53 Log CFU/g) and reduced the activity of the browning-responsible enzymes up to 50 %. During the shelf life, treated samples exhibited significantly higher scavenging activity for DPPH, ABTS, OH, O 2 - , and NO compared to non-treated samples, with minor exceptions at a high gas-to-product ratio. Additionally, treated samples showed increased levels of glucose and fructose and a comparable or higher bioaccessibility of antioxidants with respect to the products packed in air or in CO 2 . Sensory evaluation indicated that the treatment enhanced color and smell appreciation among panelists, demonstrating the potential of this method to improve both safety and quality of fresh-cut squash., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Sara Spilimbergo reports financial support was provided by Italian Ministry of Agriculture, Food Sovereignty and Forests. Krystian Marszalek reports financial support was provided by National Center for Research and Development. Zineb Benmechernene reports financial support was provided by Ministry of Higher Education and Scientific Research (Algeria). Yasin Ozdemir reports financial support was provided by General Directorate of Agricultural Research and Policies of the Ministry of Agriculture and Forestry of the Republic of Türkiye. Sara Spilimbergo reports administrative support was provided by H2020 ERA-NETs SUSFOOD2 and CORE Organic Cofunds. Simona Fabroni reports financial support was provided by Italian Ministry of University and Research. Sara Spilimbergo reports financial support was provided by Italian Ministry of Enterprises and Made in Italy. Fabio Santi reports financial support was provided by Italian Ministry of University and Research under the PNRR, Mission 4. Sara Spilimbergo has patent #IT10201700009804 issued to University of Padova. Alessandro Zambon has patent #IT10201700009804 issued to University of Padova. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

In light of the growing interest in products with reduced sugar content, there is a need to consider reducing the natural sugar concentration in juices while preserving the initial concentration of nutritional compounds. This paper reviewed the current state of knowledge related to mixing juices, membrane processes, and enzymatic processes in producing fruit juices with reduced concentrations of sugars. The limitations and challenges of these methods are also reviewed, including the losses of nutritional ingredients in membrane processes and the emergence of side products in enzymatic processes. As the existing methods have limitations, the review also identifies areas that require further improvements and technological innovations.

Identification of betacyanins in Basella alba L. and Basella alba L. var. 'Rubra' fruits was performed by low- and high-resolution mass spectrometry (LRMS and HRMS) as well as 1 H, 13 C and two-dimensional NMR which revealed hitherto completely not known betacyanin classes in the plant kingdom. Especially, the presence of unique nitrogenous acyl moieties in the structures of the pigments was ascertained by the HRMS Orbitrap detection. Except for detected polar betacyanin glycosylated derivatives, presence of a series of previously not reported pigments such as malonylated betanidin 6- O - β -glusosides with their acyl migration isomers along with the evidence of the 3''-hydroxy-butyrylated betacyanins is reported. The first complete NMR data were obtained for novel and principal acylated gomphrenins with hydroxycinnamic acids: 6'- O - E -caffeoyl-gomphrenin (malabarin), 6'- O - E -sinapoyl-gomphrenin (gandolin), 6'- O - E -4-coumaroyl-gomphrenin (globosin) and 6'- O - E -feruloyl-gomphrenin (basellin)., Competing Interests: The authors declare no conflicts of interest.

Betacyanins are a group of water-soluble red-violet compounds containing nitrogen in their structure. These are biosynthesized in red beetroot ( Beta vulgaris L.), a widely consumed vegetable that contains significant amounts of nutritious and bioactive compounds which are also found in dietary supplements. This contribution presents results of betacyanin thermal oxidation (resulting in dehydrogenation) interrelated with decarboxylation in selected acetate/phosphate buffers at pH 3-8 and at 85 °C, which may be of particular significance for formulation and performance of foods. Most of the reaction products were detected at the highest concentrations in the acidic solutions (pH 3-4). The main dehydrogenation reaction pathways were monitored by LC-DAD-MS/MS and were associated with decarboxylation of the principal extract pigments, betanin/isobetanin and neobetanin, at carbon positions C-2 and C-17. Additional reactions are accompanied by the 2,15-decarboxylation processes at different dehydrogenation levels with 15-decarboxy-betanin and 2,15-bidecarboxy-betanin, structurally elucidated by NMR analysis, as the distinct indicators of this route type. For other novel pigments detected, 2,15-bidecarboxy-xanbetanin, 2,15-bidecarboxy-xanneobetanin and 2,15,17-tridecarboxy-neobetanin, additional high resolution mass spectrometric analyses were performed and confirmed their molecular formulas.

A 49-year-old man presented with a 1-week history of abdominal pain, distension, diarrhoea and fatigue. CT of the abdomen and pelvis revealed peritonitis with no identifiable cause. Diagnostic laparoscopy was performed, which excluded gastrointestinal perforation. Peritoneal fluid tested positive for Chlamydia trachomatis and rectal swabs were positive for C. trachomatis serovars consistent with lymphogranuloma venereum (LGV). Additional blood tests also revealed a diagnosis of syphilis. This is a rare documented case of LGV peritonitis in a male without associated immunodeficiency. The patient recovered well following laparoscopic washout and a course of appropriate antibiotics., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2021. No commercial re-use. See rights and permissions. Published by BMJ.)

A 3D finite element model of the mandible dentition was developed, including 14 teeth, a periodontal ligament (PDL), and a splint made of polymethylmethacrylate (PMMA). The study considered three design options: 1-the case of splint absence; 2-the case of the splint presence installed after manufacture; and 3-the case of splint presence installed after correction (grinding) performed to ensure a uniform distribution of occlusal force between the teeth. For cases of absence and presence of splint, three measurements of the functional load were performed using the T-Scan III software and hardware complex (TekScan, Boston, MA, USA). It was found that the presence of a splint led to a decrease in the total value of the occlusive load and to a uniform distribution between all the mandible teeth. The occlusal force was considered as a static vertical force evenly distributed between the nodes belonging to the occlusive surface of the corresponding tooth for the first design option and the occlusal surface of the splint for the second and third ones, respectively. As a result of the study, it was concluded that the splint usage was effective in order to change the distribution of the functional load during the treatment of proved masticatory muscles' parafunction; the safety of using a splint for teeth and surrounding tissues under the influence of the considered functional load was shown; the potential applicability of PMMA as a structural material of a splint that had been used for the treatment of masticatory muscles' parafunction was established.

Herein, the generation of decarboxylated derivatives of gomphrenin pigments exhibiting potential health-promoting properties and the kinetics of their extraction during tea brewing from the purple flowers of Gomphrena globosa L. in aqueous and aqueous citric acid solutions were investigated. Time-dependent concentration monitoring of natural gomphrenins and their tentative identification was carried out by LC-DAD-ESI-MS/MS. The high content of acylated gomphrenins and their principal decarboxylation products, 2-, 15-, 17-decarboxy-gomphrenins, along with minor levels of their bidecarboxylated derivatives, were reported in the infusions. The identification was supported by the determination of molecular formulas of the extracted pigments by liquid chromatography coupled with high-resolution mass spectrometry (LCMS-IT-TOF). The influence of plant matrix on gomphrenins' stability and generation of their derivatives, including the extraction kinetics, was determined by studying the concentration profiles in the primary and diluted infusions. Isolated and purified acylated gomphrenins from the same plant material were used for the preliminary determination of their decarboxylated derivatives. The acylated gomphrenins were found to be more stable than nonacylated ones. Citric acid addition had a degradative influence on natural gomphrenins mainly during the longer tea brewing process (above 15 min); however, the presence of plant matrix significantly increased the stability for betacyanins' identification.

Anisotropic optical and transport properties of monolayer ReS2 fabricated by mechanical exfoliation are reported. Transient absorption measurements with different polarization configurations and sample orientations reveal that the absorption coefficient and transient absorption are both anisotropic, with maximal and minimal values occurring when the light polarization is parallel and perpendicular to the Re atomic chains, respectively. The maximal values are about a factor of 2.5 of the minimal values. By resolving the spatiotemporal dynamics of excitons, it is found that the diffusion coefficient of excitons moving along Re atomic chains is about 16 cm(2) s(-1) at room temperature, which is about a factor of three larger than those moving perpendicular to that direction. An exciton lifetime of 40 ps is also extracted. These findings establish monolayer ReS2 as an anisotropic 2D transition metal dichalcogenide., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

One issue in genome maintenance is how DNA repair proteins find lesions at rates that seem to exceed diffusion-limited search rates. We propose a phenomenon where DNA damage induces nucleosomal rearrangements which move lesions to potential rendezvous points in the chromatin structure. These rendezvous points are the dyad and the linker DNA between histones, positions in the chromatin which are more likely to be accessible by repair proteins engaged in a random search. The feasibility of this mechanism is tested by considering the statistical mechanics of DNA containing a single lesion wrapped onto the nucleosome. We consider lesions which make the DNA either more flexible or more rigid by modeling the lesion as either a decrease or an increase in the bending energy. We include this energy in a partition function model of nucleosome breathing. Our results indicate that the steady state for a breathing nucleosome will most likely position the lesion at the dyad or in the linker, depending on the energy of the lesion. A role for DNA binding proteins and chromatin remodelers is suggested based on their ability to alter the mechanical properties of the DNA and DNA-histone binding, respectively. We speculate that these positions around the nucleosome potentially serve as rendezvous points where DNA lesions may be encountered by repair proteins which may be sterically hindered from searching the rest of the nucleosomal DNA. The strength of the repositioning is strongly dependent on the structural details of the DNA lesion and the wrapping and breathing of the nucleosome. A more sophisticated evaluation of this proposed mechanism will require detailed information about breathing dynamics, the structure of partially wrapped nucleosomes, and the structural properties of damaged DNA.

An experimental and clinical study was carried out on 76 patients with implant-supported restorations in the posterior zones of the lower jaw, divided in two groups (with no signs of bone resorption around the implants and with resorption signs) and one control group. The results of the study allowed optimizing the angulation of dental implants, providing an opportunity for individual positioning of dental implant in the mandible depending on the direction of functional load.

We demonstrate a proof of concept for detecting heterogeneities and estimating lifetimes in time-correlated single-photon-counting (TCSPC) data when photon counts per molecule are low. In this approach photons are classified as either prompt or delayed according to their arrival times relative to an arbitrarily chosen time gate. Under conditions in which the maximum likelihood (ML) methods fail to distinguish between heterogeneous and homogeneous data sets, histograms of the number of prompt photons from many molecules are analyzed to identify heterogeneities, estimate the contributing fluorescence lifetimes, and determine the relative amplitudes of the fluorescence, scatter, and background components of the signal. The uncertainty of the lifetime estimate is calculated to be larger than but comparable to the uncertainty in ML estimates of single lifetime data made with similar total photon counts. Increased uncertainty and systematic errors in lifetime estimates are observed when the temporal profile of the lifetime decay is similar to either the background or scatter signals, primarily due to error in estimating the amplitudes of the various signal components. Unlike ML methods, which can fail to converge on a solution for a given molecule, this approach does not discard any data, thus reducing the potential for introducing a bias into the results., (© 2011 Society for Applied Spectroscopy)

Axiography [graphic registration of movement of a hinged (axis) condyles temporomandibular joint (ТМJ)] is an objective method of research of trajectory of the articulate movement that allows to estimate character of movements in norm and in functional disturbances of TMJ. Electronic axiographic study allowed to analyze such parametres as: quality indicators, quantity indicators (range), symmetry, synchronism of rate of movement (between left and right TMJ). The occlusion-articulation disturbances caused by a pathology of an occlusion and discoordination masseters lead chronically proceeding discoordination movements of the disk and the mandible head, as causes development of functional disturbances of TMJ.

We have previously shown that genetic variability in CNR1 is associated with low HDL dyslipidemia in a multigenerational obesity study cohort of Northern European descent (209 families, median  = 10 individuals per pedigree). In order to assess the impact of CNR1 variability on the development of dyslipidemia in the community, we genotyped this locus in all subjects with class III obesity (body mass index >40 kg/m(2)) participating in a population-based biobank of similar ancestry. Twenty-two haplotype tagging SNPs, capturing the entire CNR1 gene locus plus 15 kb upstream and 5 kb downstream, were genotyped and tested for association with clinical lipid data. This biobank contains data from 645 morbidly obese study subjects. In these subjects, a common CNR1 haplotype (H3, frequency 21.1%) is associated with fasting TG and HDL cholesterol levels (p = 0.031 for logTG; p = 0.038 for HDL-C; p = 0.00376 for log[TG/HDL-C]). The strength of this relationship increases when the data are adjusted for age, gender, body mass index, diet and physical activity. Mean TG levels were 160±70, 155±70, and 120±60 mg/dL for subjects with 0, 1, and 2 copies of the H3 haplotype. Mean HDL-C levels were 45±10, 47±10, and 48±9 mg/dL, respectively. The H3 CNR1 haplotype appears to exert a protective effect against development of obesity-related dyslipidemia.

Two complementary methods in confocal single-molecule fluorescence spectroscopy are presented to analyze conformational dynamics by Forster resonance energy transfer (FRET) measurements considering simulated and experimental data. First, an extension of photon distribution analysis (PDA) is applied to characterize conformational exchange between two or more states via global analysis of the shape of FRET peaks for different time bins. PDA accurately predicts the shape of FRET efficiency histograms in the presence of FRET fluctuations, taking into account shot noise and background contributions. Dynamic-PDA quantitatively recovers FRET efficiencies of the interconverting states and relaxation times of dynamics on the time scale of the diffusion time t(d) (typically milliseconds), with a dynamic range of the method of about +/-1 order of magnitude with respect to t(d). Correction procedures are proposed to consider the factors limiting the accuracy of dynamic-PDA, such as brightness variations, shortening of the observation time due to diffusion, and a contribution of multimolecular events. Second, an analysis procedure for multiparameter fluorescence detection is presented, where intensity-derived FRET efficiency is correlated with the fluorescence lifetime of the donor quenched by FRET. If a maximum likelihood estimator is applied to compute a mean fluorescence lifetime of mixed states, one obtains a fluorescence weighted mean lifetime. Thus a mixed state is detected by a characteristic shift of the fluorescence lifetime, which becomes longer than that expected for a single species with the same intensity-derived FRET efficiency. Analysis tools for direct visual inspection of two-dimensional diagrams of FRET efficiency versus donor lifetime are presented for the cases of static and dynamic FRET. Finally these new techniques are compared with fluorescence correlation spectroscopy.

Analysis of anisotropy in single-molecule fluorescence experiments using the probability distribution analysis (PDA) method is presented. The theory of anisotropy-PDA is an extension of the PDA theory recently developed for the analysis of Förster resonance energy transfer (FRET) signals [Antonik, M.; et al. J. Phys. Chem. B 2006, 110, 6970]. The PDA method predicts the shape of anisotropy histograms for any given expected ensemble anisotropy, signal intensity distribution, and background. Further improvements of the PDA theory allow one to work with very low photon numbers, i.e., starting from the level of background signal. Analysis of experimental and simulated data shows that PDA has the major advantage to unambiguously distinguish between shot noise broadening and broadening caused by heterogeneities in the sample. Fitting of experimental histograms yields anisotropy values of individual species, which can be directly compared with those measured in ensemble experiments. Excellent agreement between the ensemble data and the results of PDA demonstrates a good absolute accuracy of the PDA method. The precision in determination of mean values depends mainly on the total number of photons, whereas the ability of PDA to detect the presence of heterogeneities strongly depends on the time window length. In its present form PDA can be also applied to computed fluorescence parameters such as FRET efficiency and scatter-corrected fluorescence anisotropy. Extension of the PDA theory to low photon numbers makes it possible to apply PDA to dynamic systems, for which high time resolution is required. In this way PDA is developed as a sensitive tool to detect biomolecular heterogeneities in space and time.

Picosecond time-resolution fluorescence signal detection over many hours is possible using the time-correlated single photon counting (TCSPC) technique. Advanced TCSPC with clock oscillator set by the pulsed laser and data analysis provides a tool to investigate processes in single molecules on time scale from picoseconds to seconds. Optical imaging techniques combined with TCSPC allow one to study the spatial distribution of fluorescence properties in solution and on a surface. Mechanical manipulation of a single macromolecule by means of an atomic-force microscope makes it possible to detect fluorescence signal changes as a function of mechanical conformations of a fluorescent dye attached to a single DNA molecule., (Copyright 2007 Wiley-Liss, Inc.)

We establish a probability distribution analysis (PDA) method for the analysis of fluorescence resonance energy transfer (FRET) signals to determine with high precision the originating value of a shot-noise-limited signal distribution. PDA theoretical distributions are calculated explicitly including crosstalk, stochastic variations, and background and represent the minimum width that a FRET distribution must have. In this way an unambiguous distinction is made between shot-noise distributions and distributions broadened by heterogeneities. This method simultaneously and effectively extracts highly resolved information from FRET distributions. The theoretical histograms match the exact profile of histograms generated from constant transfer efficiency experimental data with a chi2 near unity. The chi2 surface suggests an ultimate level of precision with FRET of < 1% of the Förster radius. Distributions of FRET signals in donor-acceptor-labeled DNA were unambiguously identified as being broader than shot-noise variations could explain. A model describing a Gaussian distribution of distances was tested with the PDA method and demonstrated 5 A inhomogeneities due to dye motions. The capability of this method to recover quantitative information from FRET distributions has potential applications for studying molecular conformations and dynamics. Potential sources for artifacts such as acceptor photobleaching, spectrally different observation volumes, and fluctuations of the Förster radius are ruled out.

Two general strategies are introduced to identify and quantify single molecules in dilute solutions by employing a spectroscopic method for data registration and specific burst analysis, denoted multiparameter fluorescence detection (MFD). MFD uses pulsed excitation and time-correlated single-photon counting to simultaneously monitor the evolution of the eight-dimensional fluorescence information (fundamental anisotropy, fluorescence lifetime, fluorescence intensity, time, excitation spectrum, fluorescence spectrum, fluorescence quantum yield, distance between fluorophores) in real time and allows for selection of specific events for subsequent analysis. Using the multiple fluorescence dimensions, we demonstrate a dye labeling scheme of oligonucleotides, by which it is possible to identify and separate 16 different compounds in the mixture via their characteristic pattern by MFD. Such identification procedures and multiplex assays with single-molecule sensitivity may have a great impact on screening of species and events that do not lend themselves so easily to amplification, such as disease-specific proteins and their interactions.

Knowledge of the optical properties of atomic force microscopy (AFM) tips is relevant for the combination of optical and force spectroscopy. The luminescence properties of five commercial AFM tips were characterized using a combination of multiparameter fluorescence detection (MFD) and scanning confocal techniques. These include three Si3N4 tips, one silicon tip, and one high-density carbon (HDC) tip grown on top of a silicon tip. Time-decay histograms of the signal were analyzed to determine the strength of scatter, constant background, and fluorescence in the observed signal. Intensity and anisotropy images with optical resolution down to the diffraction limit were generated. The optical signal recorded from the apex of the Si3N4 tips ranged from 0.7 to 1.9 times the count rates from single Rhodamine 110 molecules under similar illumination conditions. The signal is predominantly composed of scatter and background (>85%), plus a small fluorescence component with lifetimes between 1 and 3 ns. The intensity of the recorded signal fell with increasing distance from the apex, and by 300 nm the signals fell below single-molecule levels for all Si3N4 cantilevers. Silicon cantilevers demonstrated very low count rates relative to single-molecule measurements under all conditions, and virtually no fluorescence. The high-density carbon tips also demonstrated low count rates, but the signal contained a short lifetime fluorescence component (0.7 ns). The intensity of the signals from each of the tips was geometry dependent, demonstrating the highest intensities at the edges and corners. Likewise, the anisotropy of all tip signals was observed to be geometry dependent, with the dependence varying on a case-by-case basis. The implications for using confocal illumination instead of total internal reflection are discussed.