Effect of High Hydrostatic Pressure on the Extractability and Bioaccessibility of Carotenoids and Their Esters from Papaya (Carica papaya L.) and Its Impact on Tissue Microstructure

High hydrostatic pressure (HHP) is a non-thermal technology widely used in the industry to extend food shelf-life and it has been proven to enhance the extractability of secondary metabolites, such as carotenoids, in plant foods. In this study, fresh-cut papaya pulp of varieties (Sweet Mary, Alicia and Eksotika) from the Canary Islands (Spain) were submitted to the HHP process (pressure: 100, 350 and 600 MPa; time: come-up time (CUT) and 5 min) to evaluate, for the first time, individual carotenoid and carotenoid ester extractability and to assess their bioaccessibility using an in vitro simulated gastrointestinal digestion assay, following the standardized INFOGEST® methodology. In addition, changes in papaya pulp microstructure after HHP treatments and during the different phases of the in vitro digestion were evaluated with optical light microscopy. HPLC-DAD (LC-MS/MS (APCI+)) analyses revealed that HHP treatments increased the carotenoid content, obtaining the highest extractability in pulp of the Sweet Mary papaya variety treated at 350 MPa during 5 min (4469 ± 124 μg/100 g fresh weight) which was an increase of 269% in respect to the HHP-untreated control sample. The highest carotenoid extraction value within each papaya variety among all HHP treatments was observed for (all-E)-lycopene, in a range of 98–1302 μg/100 g fresh weight (23–344%). Light micrographs of HHP-treated pulps showed many microstructural changes associated to carotenoid release related to the observed increase in their content. Carotenoids and carotenoid esters of papaya pulp submitted to in vitro digestion showed great stability; however, their bioaccessibility was very low due to the low content of fatty acids in papaya pulp necessary for the micellarization process. Further studies will be required to improve papaya carotenoid and carotenoid ester bioaccessibility.

Long-Term Corrosion Testing of Zy-4 in a LiOH Solution under High Pressure and Temperature Conditions

The fuel cladding is one of the most important structural components for maintaining the integrity of a fuel channel and for safely exploitation of a nuclear power plant. The corrosion behavior of a fuel cladding material, Zy-4, under high pressure and temperatures conditions, was analyzed in a static isothermal autoclave under simulated primary water conditions—a LiOH solution at 310 °C and 10 MPa for up to 3024 h. After this, the oxides grown on the Zy-4 sample surface were characterized using electrochemical measurements, gravimetric analysis, metallographic analysis, SEM and XPS. The maximum oxide thicknesses evaluated by gravimetric and SEM measurements were in good agreement; both values were around 1.2 µm. The optical light microscopy (OLM) investigations identified the presence of small hydrides uniformly distributed horizontally across the alloy. EIS impedance spectra showed an increase in the oxide impedance for the samples oxidized for a long time. EIS plots has the best fit with an equivalent circuit which illustrated an oxide model that has two oxide layers: an inner oxide layer and outer layer. The EIS results showed that the inner layer was a barrier layer, and the outer layer was a porous layer. Potentiodynamic polarization results demonstrated superior corrosion resistance of the samples tested for longer periods of time. By XPS measurements we identified all five oxidation states of zirconium: Zr0 located at 178.5 eV; Zr4+ at 182.8 eV; and the three suboxides, Zr+, Zr2+ and Zr3+ at 179.7, 180.8 and 181.8 eV, respectively. The determination of Vickers microhardness completed the investigation.

Mesoscale Process Modeling of a Thick Pultruded Composite with Variability in Fiber Volume Fraction

Pultruded fiber-reinforced polymer composites are susceptible to microstructural nonuniformity such as variability in fiber volume fraction (Vf), which can have a profound effect on process-induced residual stress. Until now, this effect of non-uniform Vf distribution has been hardly addressed in the process models. In the present study, we characterized the Vf distribution and accompanying nonuniformity in a unidirectional fiber-reinforced pultruded profile using optical light microscopy. The identified nonuniformity in Vf was subsequently implemented in a mesoscale thermal–chemical–mechanical process model, developed explicitly for the pultrusion process. In our process model, the constitutive material behavior was defined locally with respect to the corresponding fiber volume fraction value in different-sized representative volume elements. The effect of nonuniformity on the temperature and cure degree evolution, and residual stress was analyzed in depth. The results show that the nonuniformity in fiber volume fraction across the cross-section increased the absolute magnitude of the predicted residual stress, leading to a more scattered residual stress distribution. The observed Vf gradient promotes tensile residual stress at the core and compressive residual stress at the outer regions. Consequently, it is concluded that it is essential to take the effects of nonuniformity in fiber distribution into account for residual stress estimations, and the proposed numerical framework was found to be an efficient tool to study this aspect.

Criterion validity of ultrasound in the identification of calcium pyrophosphate crystal deposits at the knee: an OMERACT ultrasound study

ObjectiveTo evaluate the discriminatory ability of ultrasound in calcium pyrophosphate deposition disease (CPPD), using microscopic analysis of menisci and knee hyaline cartilage (HC) as reference standard.MethodsConsecutive patients scheduled for knee replacement surgery, due to osteoarthritis (OA), were enrolled. Each patient underwent ultrasound examination of the menisci and HC of the knee, scoring each site for presence/absence of CPPD. Ultrasound signs of inflammation (effusion, synovial proliferation and power Doppler) were assessed semiquantitatively (0–3). The menisci and condyles, retrieved during surgery, were examined microscopically by optical light microscopy and by compensated polarised microscopy. CPPs were scored as present/absent in six different samples from the surface and from the internal part of menisci and cartilage. Ultrasound and microscopic analysis were performed by different operators, blinded to each other’s findings.Results11 researchers from seven countries participated in the study. Of 101 enrolled patients, 68 were included in the analysis. In 38 patients, the surgical specimens were insufficient. The overall diagnostic accuracy of ultrasound for CPPD was of 75%—sensitivity of 91% (range 71%–87% in single sites) and specificity of 59% (range 68%–92%). The best sensitivity and specificity were obtained by assessing in combination by ultrasound the medial meniscus and the medial condyle HC (88% and 76%, respectively). No differences were found between patients with and without CPPD regarding ultrasound signs of inflammation.ConclusionUltrasound demonstrated to be an accurate tool for discriminating CPPD. No differences were found between patents with OA alone and CPPD plus OA regarding inflammation.

AB0060 STING AND PROINFLAMMATORY CYTOKINES IN SYNOVIAL FLUID OF PATIENTS WITH DIFFERENT ARTHRITIDES

Background:STING (stimulator of interferon genes) is a cytosolic protein that is found in endoplasmic reticulum (ER) membrane, mitochondria and mitochondria-associated membranes. Although it is well established that STING plays an important role in innate immune responses, its potential involvement in rheumatic disease processes remains to be clarified (1).Objectives:The aims of this study were to evaluate the levels of STING and its relationship with local inflammation in the synovial fluid (SF) of patients with psoriatic arthritis (PsA), rheumatoid arthritis (RA), gout, calcium pyrophosphate (CPP) crystal-induced arthritis (CPP-IA), osteoarthritis (OA) and OA with CPP crystals (OA+CPP).Methods:SF was collected from the knees of 60 untreated patients: 10 with PsA, 10 with RA, 10 with gout, 10 with CPP-IA, 10 with OA and 10 with OA+CPP. SF was examined under optical light microscopy. White cell count (WBC) and the polymorphonuclear cell (PMN) percentage were determined in SF according to standard procedures. SF IL-8, IL-6, IL-1β and extra- and intra-cellular STING levels were assayed by ELISA.Results:The levels of WBC were higher in SFs from gouty patients (27.7±20.56 103/mm3), while OA and OA+CPP patients showed the lowest WBC count (0.34±0.3 103/mm3, 0.3±0.28 103/mm3). SFs from inflammatory arthritis contained elevated percentages of PMN (gout: 85.5±10.86%, CPP-IA: 84±11.31%, RA: 80.33±8.14%, PsA: 42.6±35.97%). Extracellular STING was determined in OA (440±413.31 pg/ml), OA+CPP (225±205.06 pg/ml) and CPP-IA (475±7.07 pg/ml) SF, while was not detectable in RA, PsA and gout. Intracellular STING levels were similar and the highest in SFs from gout (96.4±35.44 pg/ml) and RA (90.64±23.13 pg/ml), while remained under detection limit only in SFs from PsA. SF concentration of IL-6 was lower in OA (354.87±377.56 pg/ml) and OA+CPP (389.56±104.14 pg/ml) as compared with inflammatory arthritides (PsA: 3807.14±489.86 pg/ml; RA: 17354±2334.87 pg/ml; gout: 19359±84.85 pg/ml; CPP-IA: 20389.56±104.14 pg/ml). The patients with gout and RA had the highest levels of IL-8 (2159.54±347.09 pg/ml; 2039.6±97.74 pg/ml) and IL-1β (35.93±20.46 pg/ml; 44.36±23.16 pg/ml), while OA showed the lowest concentrations (IL-8: 23.21±11.32 pg/ml; IL-1β: 0.47±0.13 pg/ml). In the total group of patients, we found a negative correlation between extracellular STING and IL-8 (r=-0.53; p=0.004) and IL-1β (r=-0.47; p=0.012). There was a positive correlation between intracellular STING and IL-8 (r=0.54; p=0.017), IL-1β (r=0.77; p<0.001) and IL-6 (r=0.69 p=0.009).Conclusion:This study is the first to define the presence of STING in SF of different arthritides. The high levels of extracellular STING in OA, OA+CPP and CPP-IA SFs may be due to the activation of factors that reduce its interaction with the ER. The effect of downregulating factors in PsA might explain the low concentration of intracellular STING in these patients.References:[1]Barber GN. STING: infection, inflammation and cancer. Nat Rev Immunol 2015;15:760-70.Disclosure of Interests:Anna Scanu: None declared, Roberto Luisetto: None declared, Francesca Oliviero: None declared, Paola Galozzi: None declared, Augusta Ortolan: None declared, Mariagrazia Lorenzin: None declared, Mara Felicetti: None declared, Andrea Doria Consultant of: GSK, Pfizer, Abbvie, Novartis, Ely Lilly, Speakers bureau: UCB pharma, GSK, Pfizer, Janssen, Abbvie, Novartis, Ely Lilly, BMS, Roberta Ramonda Speakers bureau: Novartis, Celgene, Janssen, Pfizer, Abbvie, Lilly

Development of an artificial intelligence-based assessment model for prediction of embryo viability using static images captured by optical light microscopy during IVF

STUDY QUESTION Can an artificial intelligence (AI)-based model predict human embryo viability using images captured by optical light microscopy? SUMMARY ANSWER We have combined computer vision image processing methods and deep learning techniques to create the non-invasive Life Whisperer AI model for robust prediction of embryo viability, as measured by clinical pregnancy outcome, using single static images of Day 5 blastocysts obtained from standard optical light microscope systems. WHAT IS KNOWN ALREADY Embryo selection following IVF is a critical factor in determining the success of ensuing pregnancy. Traditional morphokinetic grading by trained embryologists can be subjective and variable, and other complementary techniques, such as time-lapse imaging, require costly equipment and have not reliably demonstrated predictive ability for the endpoint of clinical pregnancy. AI methods are being investigated as a promising means for improving embryo selection and predicting implantation and pregnancy outcomes. STUDY DESIGN, SIZE, DURATION These studies involved analysis of retrospectively collected data including standard optical light microscope images and clinical outcomes of 8886 embryos from 11 different IVF clinics, across three different countries, between 2011 and 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS The AI-based model was trained using static two-dimensional optical light microscope images with known clinical pregnancy outcome as measured by fetal heartbeat to provide a confidence score for prediction of pregnancy. Predictive accuracy was determined by evaluating sensitivity, specificity and overall weighted accuracy, and was visualized using histograms of the distributions of predictions. Comparison to embryologists’ predictive accuracy was performed using a binary classification approach and a 5-band ranking comparison. MAIN RESULTS AND THE ROLE OF CHANCE The Life Whisperer AI model showed a sensitivity of 70.1% for viable embryos while maintaining a specificity of 60.5% for non-viable embryos across three independent blind test sets from different clinics. The weighted overall accuracy in each blind test set was &gt;63%, with a combined accuracy of 64.3% across both viable and non-viable embryos, demonstrating model robustness and generalizability beyond the result expected from chance. Distributions of predictions showed clear separation of correctly and incorrectly classified embryos. Binary comparison of viable/non-viable embryo classification demonstrated an improvement of 24.7% over embryologists’ accuracy (P = 0.047, n = 2, Student’s t test), and 5-band ranking comparison demonstrated an improvement of 42.0% over embryologists (P = 0.028, n = 2, Student’s t test). LIMITATIONS, REASONS FOR CAUTION The AI model developed here is limited to analysis of Day 5 embryos; therefore, further evaluation or modification of the model is needed to incorporate information from different time points. The endpoint described is clinical pregnancy as measured by fetal heartbeat, and this does not indicate the probability of live birth. The current investigation was performed with retrospectively collected data, and hence it will be of importance to collect data prospectively to assess real-world use of the AI model. WIDER IMPLICATIONS OF THE FINDINGS These studies demonstrated an improved predictive ability for evaluation of embryo viability when compared with embryologists’ traditional morphokinetic grading methods. The superior accuracy of the Life Whisperer AI model could lead to improved pregnancy success rates in IVF when used in a clinical setting. It could also potentially assist in standardization of embryo selection methods across multiple clinical environments, while eliminating the need for complex time-lapse imaging equipment. Finally, the cloud-based software application used to apply the Life Whisperer AI model in clinical practice makes it broadly applicable and globally scalable to IVF clinics worldwide. STUDY FUNDING/COMPETING INTEREST(S) Life Whisperer Diagnostics, Pty Ltd is a wholly owned subsidiary of the parent company, Presagen Pty Ltd. Funding for the study was provided by Presagen with grant funding received from the South Australian Government: Research, Commercialisation and Startup Fund (RCSF). ‘In kind’ support and embryology expertise to guide algorithm development were provided by Ovation Fertility. J.M.M.H., D.P. and M.P. are co-owners of Life Whisperer and Presagen. Presagen has filed a provisional patent for the technology described in this manuscript (52985P pending). A.P.M. owns stock in Life Whisperer, and S.M.D., A.J., T.N. and A.P.M. are employees of Life Whisperer.