Autologous Microfragmented Adipose Tissue Reduces the Catabolic and Fibrosis Response in an in Vitro Model of Tendon Cell Inflammation

Background. Mesenchymal stem cells (MSCs) emerged as a promising therapy for tendon pathologies. Microfragmented adipose tissue (\u3bcFAT) represents a convenient autologous product for the application of MSC-based therapies in the clinical setting. In the present study, the ability of \u3bcFAT to counteract inflammatory processes induced by IL-1\u3b2 on human tendon cells (TCs) was evaluated. Methods. Cell viability and proliferation were evaluated after 48 hours of transwell coculture of TCs and autologous \u3bcFAT in the presence or absence of IL-1\u3b2. Gene expression of scleraxis, collagen type I and type III, metalloproteinases-1 and -3, and cyclooxygenase-2 was evaluated by real-time RT-PCR. The content of VEGF, IL-1Ra, TNF\u3b1, and IL-6 was evaluated by ELISA. Results. IL-1\u3b2-treated TCs showed augmented collagen type III, metalloproteases, and cyclooxygenase-2 expression. \u3bcFAT was able to reduce the expression of collagen type III and metalloproteases-1 in a significant manner, and at the same time, it enhanced the production of VEGF, IL-1Ra, and IL-6. Conclusions. In this in vitro model of tendon cell inflammation, the paracrine action of \u3bcFAT, exerted by anti-inflammatory molecules and growth factors, was able to inhibit the expression of fibrosis and catabolic markers. Then, these results suggest that the application of \u3bcFAT may represent an effective conservative or adjuvant therapy for the treatment of tendon disorders

Osmotic dehydration of organic kiwifruit pre-treated by pulsed electric fields and monitored by NMR

[EN] Osmotic dehydration (OD) is a widely used preservation technique that consists in the reduction in food water activity by the immersion of the biological tissue in hypertonic solutions. The aim of this work was to analyze the effect of pulsed electric fields (PEF) in mass transfer as a pre-treatment of the OD using NMR. In this sense, PEF pre-treatments were done using three different voltages (100, 250 and 400 V/cm) and 60 number of pulse. The OD of kiwifruit was carried out in 61.5% of sucrose solution at 25 °C, for a contact period from 0 to 120 min. The water distribution into the cellular tissue was studied by NMR relaxometry. In conclusion, NMR is an excellent technique for quantifying water molecules according to their interactions in the fruit tissue, obtaining the adsorbed water and opening the possibility to apply the BET model to fit the adsorbed isotherm over the whole range of water activity.The authors Urszula Tylewicz and Marco Dalla Rosa want to thank for the financial support provided by funding bodies within the FP7 ERA-Net CORE Organic Plus, and with cofounds from the European Commission. The author Maria Victoria Traffano Schiffo wants to thank the FPI Predoctoral Program of the Universidad Politecnica de Valencia for support her PhD studies, ERASMUS PRACTICAS program to finance her mobility to Italy. The authors Pedro J. Fito, Marta Castro-Giraldez and M. Victoria Traffano-Schiffo acknowledge the financial support from the Spanish Ministerio de Economia, Industria y Competitividad, Programa Estatal de I+D+i orientada a los Retos de la Sociedad AGL2016-80643-R.Traffano-Schiffo, MV.; Laghi, L.; Castro Giráldez, M.; Tylewicz, U.; Rocculi, P.; Ragni, L.; Dalla Rosa, M.... (2017). Osmotic dehydration of organic kiwifruit pre-treated by pulsed electric fields and monitored by NMR. Food Chemistry. 236:87-93. https://doi.org/10.1016/j.foodchem.2017.02.046S879323

Osmotic dehydration of organic kiwifruit pre-treated by pulsed electric fields: Internal transport and transformations analyzed by NMR

[EN] This work analyzes the effect of Pulsed Electric Fields (PEF) as a pre-treatment of the osmotic dehydration (OD) of kiwifruit (Actinidia deliciosa cv Hayward) in the internal structure and in the internal water transport. PEF pre-treatments were done using three PEF intensities (100, 250 and 400 V/cm) and analyzed by TD-NMR. The OD was carried out by immersing the samples in 61.5% sucrose solution at 25 °C. The application of a PEF pre-treatment before the OD produces a process of plasmolysis proportional to the electric field applied. It is because the PEF removes the mobile charges of the medium, such as electrolytes, organic acids, amino acids; Ca+2 is the major culprit of the plasmolysis because it fixes some of the junctions of the microtubules between the cell wall and the membrane. Therefore, a previous plasmolysis produces an increase in the apoplastic transport increasing the rate of dehydration.The authors Urszula Tylewicz and Marco Dalla Rosa want to thank for the financial support provided by funding bodies within the FP7 ERA-Net CORE Organic Plus, and with cofounds from the European Commission. The author Maria Victoria Traffano Schiffo wants to thank the FPI Predoctoral Program of the Universidad Politecnica de Valencia for support her PhD studies, ERASMUS PRACTICAS program to finance her mobility to Italy. The authors Pedro J. Fito, Marta Castro-Giraldez and M. Victoria Traffano-Schiffo acknowledge the financial support from the Spanish Ministerio de Economia, Industria y Competitividad, Programa Estatal de I + D + i orientada a los Retos de la Sociedad AGL2016-80643-R, Agencia Estatal de Investigacion (AEI) and Fondo Europeo de Desarrollo Regional (FEDER).Traffano-Schiffo, MV.; Laghi, L.; Castro Giráldez, M.; Tylewicz, U.; Romani, S.; Ragni, L.; Dalla Rosa, M.... (2017). Osmotic dehydration of organic kiwifruit pre-treated by pulsed electric fields: Internal transport and transformations analyzed by NMR. Innovative Food Science & Emerging Technologies. 41:259-266. https://doi.org/10.1016/j.ifset.2017.03.012S2592664

Effect of pulsed electric field pre-treatment on microstructure and internal transport throughout osmotic treatment of organic kiwifruit.

Plant cellular structure could be considered a complex organized system where flows are carried out by different solutes or solvents transports systems. Passive transports, which is based on the free energy gradients, involve symplastic, apoplastic and aquaporins transmembrane transports. During osmotic dehydration (OD) treatment the semipermeable membranes such as plasma membrane and tonoplast, are forced to separate, due to the water losses from the vacuoles, starting the process known as plasmolysis, characterized by the loss of the turgor pressure. In the last years, OD has been extensively studied for the partial dehydration of fruits and vegetables in order to obtain semi-moist products; however, it presents some limitations such as the low dehydration rate and the high solute content in the final product. Therefore, the use of pre-treatment such as Pulsed Electric Fields (PEF) has been reported to facilitate water removal and to improve the quality of the dried or osmo-dried products. PEF is a non-thermal technology which involves the application of short and repeated voltage pulses to a biological tissue placed between two electrodes; it induces changes and reorganization in the electric conformation of the cell membrane, modifying the normal fluxes during drying process when it is used as a pre-treatment. In present work PEF has been applied as a pre-treatment prior OD of organic kiwifruits (Actinidia deliciosa cv Hayward) in order to evaluate its effect on the internal structure and internal water transport. PEF pre-treatments were performed using the following parameters: E= 100, 250 and 400 V/cm, 60 near-rectangular shape pulses, pulse width of 100 ± 2 μs and a repetition time of 10.0 ± 0.1 ms. The OD was carried out by immersing the samples in 61.5% sucrose solution at 25 °C for different time period (0-120 min). The samples were analyzed in terms of microstructure by Cryo-SEM microscopy and internal water transport by Time Domain Nuclear Magnetic Resonance (TD-NMR). The results showed that the application of a PEF pre-treatment before the OD produces a process of plasmolysis proportional to the electric field strength applied. It is because the PEF removes the mobile charges of the medium, such as electrolytes, organic acids, amino acids; Ca+2 is the major culprit of the plasmolysis because it fixes some of the junctions of the microtubules between the cell wall and the membrane. In addition, the process of plasmolysis induced by the electric field changes the behavior of kiwifruit tissue during the OD process. In a standard OD without any pretratment, the main transport is the symplastic, whereas if previously treated with PEF, the apoplastic transport is as important as the symplastic, considerably increasing the rate of dehydration. Acknowledgements: Financial support for this project is provided by funding bodies within the FP7 ERA-Net CORE Organic Plus, and with cofounds from the European Commission (No 618107)

[¹⁸F]FMCH PET/CT biomarkers and similarity analysis to refine the definition of oligometastatic prostate cancer

Background:The role of image-derived biomarkers in recurrent oligometastatic Prostate Cancer (PCa) is unexplored. This paper aimed to evaluate [18F]FMCH PET/CT radiomic analysis in patients with recurrent PCa after primary radical therapy. Specifically, we tested intra-patient lesions similarity in oligometastatic and plurimetastatic PCa, comparing the two most used definitions of oligometastatic disease.Methods:PCa patients eligible for [18F]FMCH PET/CT presenting biochemical failure after first-line curative treat-ments were invited to participate in this prospective observational trial. PET/CT images of 92 patients were visually and quantitatively analyzed. Each patient was classified as oligometastatic or plurimetastatic according to the total number of detected lesions (up to 3 and up to 5 or > 3 and > 5, respectively). Univariate and intra-patient lesions’ similarity analysis were performed.Results: [18F]FMCH PET/CT identified 370 lesions, anatomically classified as regional lymph nodes and distant metastases. Thirty-eight and 54 patients were designed oligometastatic and plurimetastatic, respectively, using a 3-lesion threshold. The number of oligometastic scaled up to 60 patients (thus 32 plurimetastatic patients) with a 5-lesion threshold. Similarity analysis showed high lesions’ heterogeneity. Grouping patients according to the number of metastases, patients with oligometastatic PCa defined with a 5-lesion threshold presented lesions heterogene-ity comparable to plurimetastic patients. Lesions within patients having a limited tumor burden as defined by three lesions were characterized by less heterogeneity.Conclusions:We found a comparable heterogeneity between patients with up to five lesions and plurimetastic patients, while patients with up to three lesions were less heterogeneous than plurimetastatic patients, featuring dif-ferent cells phenotypes in the two groups. Our results supported the use of a 3-lesion threshold to define oligometa-static PCa

Insights from cardiopulmonary exercise testing in pediatric patients with hypertrophic cardiomyopathy

The usefulness of cardiopulmonary exercise test (CPET) in adult hypertrophic cardiomyopathy (HCM) patients is well-known, whereas its role in pediatric HCM patients has not yet been explored. The present study investigates possible insights from a CPET assessment in a cohort of pediatric HCM outpatients in terms of functional and prognostic assessment. Sixty consecutive pediatric HCM outpatients aged <18 years old were enrolled, each of them undergoing a full clinical assessment including a CPET; a group of 60 healthy subjects served as controls. A unique composite end-point of heart failure (HF) related and sudden cardiac death (SCD) or SCD-equivalent events was also explored. During a median follow-up of 53 months (25th–75th: 13–84 months), a total of 13 HF- and 7 SCD-related first events were collected. Compared to controls, HCM patients showed an impaired functional capacity with most of them showing peak oxygen uptake (pVO2) values of <80% of the predicted, clearly discrepant with functional New York Heart Association class assessment. The composite end-point occurred more frequently in patients with the worst CPETs’ profiles. At the univariate analysis, pVO2% was the variable with the strongest association with adverse events at follow-up (C-index = 0.72, p = 0.025) and a cut-off value equal to 60% was the most accurate in identifying those patients at the highest risk. In a pediatric HCM subset, the CPET assessment allows a true functional capacity estimation and it might be helpful in identifying early those patients at high risk of events.publishedVersio

Moderate and Severe Congenital Heart Diseases Adversely Affect the Growth of Children in Italy: A Retrospective Monocentric Study

: Children with congenital heart disease (CHD) are at increased risk for undernutrition. The aim of our study was to describe the growth parameters of Italian children with CHD compared to healthy children. We performed a cross-sectional study collecting the anthropometric data of pediatric patients with CHD and healthy controls. WHO and Italian z-scores for weight for age (WZ), length/height for age (HZ), weight for height (WHZ) and body mass index (BMIZ) were collected. A total of 657 patients (566 with CHD and 91 healthy controls) were enrolled: 255 had mild CHD, 223 had moderate CHD and 88 had severe CHD. Compared to CHD patients, healthy children were younger (age: 7.5 ± 5.4 vs. 5.6 ± 4.3 years, p = 0.0009), taller/longer (HZ: 0.14 ± 1.41 vs. 0.62 ± 1.20, p &lt; 0.002) and heavier (WZ: -0,07 ± 1.32 vs. 0.31 ± 1.13, p = 0.009) with no significant differences in BMIZ (-0,14 ± 1.24 vs. -0.07 ± 1.13, p = 0.64) and WHZ (0.05 ± 1.47 vs. 0.43 ± 1.07, p = 0.1187). Moderate and severe CHD patients presented lower z-scores at any age, with a more remarkable difference in children younger than 2 years (WZ) and older than 5 years (HZ, WZ and BMIZ). Stunting and underweight were significantly more present in children affected by CHD (p &lt; 0.01). In conclusion, CHD negatively affects the growth of children based on the severity of the disease, even in a high-income country, resulting in a significant percentage of undernutrition in this population

488 Candidacy for heart transplantation in adult congenital heart disease patients: a single-centre, retrospective, cohort study

Abstract Aims End-stage heart failure (HF) is the leading cause of death in adult congenital heart disease (ACHD) population. Heart transplantation (HTx) improves prognosis in ACHD end-stage HF but candidacy evaluation, referral pattern, and correct listing timing are not fully elucidated in this population. To evaluate factors associated to refusal from Htx in ACHD patients with end-stage HF referred for HTx evaluation. Methods and results This retrospective cohort study enrolled consecutive ACHD patients considered for HTx in our institution between 2014 and 2020 and patients undergone HTx between 2000 and 2013. Refusal from HTx served as primary study endpoint. Between 2014 and 2020, 46 ACHD patients were evaluated for HTx, 14 ACHD patients underwent HTx between 2001 and 2013. The main indication to HTx in patients with single ventricle physiology was Fontan failure, while in patients with systemic left ventricle and systemic right ventricle physiology, it was systemic ventricular dysfunction. We compared clinical, anatomical and demographic data of 41 patients accepted for transplantation with 15 patients refused after screening. Risk factors for refusal were: coexistence of multiple high risk features [odds ratio (OR): 3.6; 95% confidence interval (CI): 1.1–12.9; P 0.048]; anatomical factors (OR: 14.5; 95% CI: 3.1–68.4; P 0.001), out-of-centre ACHD/HTx program referral (OR: 5.3; 95% CI: 1.5 to 19.0; p 0.01). Survival in patients accepted for HTx was significantly higher than survival in patients declined from HTx with landmark comparison at 20, 40 and 60 months of 87%, 78%, and 72% vs. 70%, 59%, and 20%, respectively. HTx refusal identifies a high risk ACHD patient subgroup (hazard ratio for overall mortality: 3.1; 95% CI: 1.1–8.3; P 0.02). Conclusions In our study risk factors for refusal from HTx are adverse anatomical features, coexistence of multiple conventional HTx high risk factors and out-of-centre referral. ACHD patients refused from HTx present shorter time to death. Efforts to increase HTx candidacy and to reduce referral delay in tertiary centre are strongly necessary for this growing population

Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology

Spontaneous Raman microscopy reveals the chemical composition of a sample in a label-free and non-invasive fashion by directly measuring the vibrational spectra of molecules. However, its extremely low cross section prevents its application to fast imaging. Stimulated Raman scattering (SRS) amplifies the signal by several orders of magnitude thanks to the coherent nature of the nonlinear process, thus unlocking high-speed microscopy applications that provide analytical information to elucidate biochemical mechanisms with subcellular resolution. Nevertheless, in its standard implementation, narrowband SRS provides images at only one frequency at a time, which is not sufficient to distinguish constituents with overlapping Raman bands. Here, we report a broadband SRS microscope equipped with a home-built multichannel lock-in amplifier simultaneously measuring the SRS signal at 32 frequencies with integration time down to 44 μs, allowing for detailed, high spatial resolution mapping of spectrally congested samples. We demonstrate the capability of our microscope to differentiate the chemical constituents of heterogeneous samples by measuring the relative concentrations of different fatty acids in cultured hepatocytes at the single lipid droplet level and by differentiating tumor from peritumoral tissue in a preclinical mouse model of fibrosarcoma