ANTIMICROBIAL PEPTIDES ARE TARGETS FOR BACTERIAL ADP-RIBOSYLTRANSFERASE ENZYMES

Background: Human α-defensins (HNPs 1-3) are small cationic, amphipatic peptides with microbicidal activities. HNP-1 is the physiological target of ART1, an arginine (Arg) specific eucaryotic ribosyltransferase enzyme1. Mono ADP-ribosylation of Arg14 of HNP-1 modulates its biological activities2. Bacterial exotoxins like Cholera Toxin (CT) by Vibrio cholerae, Heat Labile Enterotoxin (LT1) by Escherichia coli and Exoenzyme S (ExoS) by Pseudomonas aeruginosa are arginine ADP-ribosyltransferase enzymes that alter cell functions by modifying protein targets. Objectives: 1. Evaluate the ADP ribosylation of HNP-1 by CT, LT1 and ExoS. 2. Purify modified peptides and identify the ADP-ribosylated Arg. Methods: ADP-ribosylation of HNP-1 will be evaluated with biotinilated NAD by western blot. Purification of modified peptides will be performed by reverse phase HPLC. Identification of modification will be performed by Maldi-Toff analysis. Results: 1. CT and LT1 are effective in ADP-ribosylating HNP-1 as equal as the well known activity of ART1. On the other hand ExoS does not recognize HNP-1 as substrate. 2. Ongoing experiments are purifications and characterization of modified peptides by reverse-phase HPLC and Maldi-Toff analysis. Conclusions: 1. The different ADP-ribosylating activities displayed by CT, LT1 and ExoS on HNP-1 might be explained with differences in microbial pathogenesis, as the toxins released by V. cholerae and E. coli are involved in the early stages of infections, during the interactions with surface epithelial cells, while ExoS by P. aeruginosa is active during blood dissemination, when the pathogen has already overcome epithelial barrier. References: 1 Balducci et al., 1999, Am J Respir Cell Mol Biol, 21, 337-46 2 Paone et al., 2006, J Biol Chem, 281, 17054-6

Kiwifruit waste valorisation through innovative snack development

Currently, in the case of kiwifruits, those fruit with a weight lower than 65 g are considered waste. The production of dried snacks with high nutritional functionality could be a valid alternative to use the kiwifruit waste, with positive economic impact on the entire production chain. Therefore, the aim of this work was to evaluate the effect of pulsed electric field – PEF (200 V cm-1) and/or osmotic dehydration – OD (trehalose at 40%) pre-drying treatments on drying kinetics at 50, 60, and 70°C, and on colour and nutritional properties (vitamin C and antioxidant compounds) of ‘Jintao’ (yellow-fleshed) kiwifruit snacks. At every temperature, the PEF treated snacks showed the highest drying rate. Moreover, PEF treatment appeared to be a valid innovative alternative for the production of fruit snacks with high nutritional quality. A better retention of vitamin C and antioxidant compounds was obtained in dried yellow kiwifruit subjected to PEF treatment

Reduction of colonic inflammation in HLA-B27 transgenic rats by feeding Marie Ménard apples, rich in polyphenols

Inflammatory bowel diseases (IBD) are immunomediated ailments affecting millions of individuals. Although diet is regarded as an important factor influencing IBD, there are no accepted dietary recommendations presently available. We administered 7.6 % lyophilised apples obtained from two cultivars (Golden Delicious and Marie Ménard, low and high in polyphenols, respectively) to HLA-B27 transgenic rats which develop spontaneous IBD. After 3 months feeding, rats fed Marie Ménard apples had reduced myeloperoxidase activity (3.6 (sem 0.3) v. 2.2 (sem 0.2) U/g tissue; P <0.05) and reduced cyclo-oxygenase-2 (P <0.05) and inducible NO synthase gene expression (P <0.01) in the colon mucosa and significantly less diarrhoea (P <0.05), compared with control rats. Cell proliferation in the colon mucosa was reduced significantly by feeding Golden Delicious apples, with a borderline effect of Marie Ménard apples. Gene expression profiling of the colon mucosa, analysed using the Whole Rat Genome 4 x 44 K Agilent Arrays, revealed a down-regulation of the pathways of PG synthesis, mitogen-activated protein kinase (MAPK) signalling and TNFalpha-NF-kappaB in Marie Ménard-fed rats. In the stools of the animals of this group we also measured a significant reduction of bacteria of the Bacteriodes fragilis group. In conclusion, the administration of Marie Ménard apples, rich in polyphenols and used at present only in the manufacturing of cider, ameliorates colon inflammation in transgenic rats developing spontaneous intestinal inflammation, suggesting the possible use of these and other apple varieties to control inflammation in IBD patient

The Impact of Plasma Activated Water Treatment on the Phenolic Profile, Vitamins Content, Antioxidant and Enzymatic Activities of Rocket-Salad Leaves

Plasma activated water (PAW) recently received much attention as an alternative food preservation method. However, its effects on food quality are still scarce. This study evaluates the effect of PAW processing time on bioactive compounds of rocket-salad leaves including: 18 phenolic compounds, vitamin C, riboflavin, nicotinic acid, and nicotinamide. Moreover, the impact of PAW on both antioxidant (DPPH) and peroxidase (POD) activities was also investigated. This was performed using HPLC-DAD, HPLC-MS/MS, and spectrophotometric analysis. All treatments induced non-significant increases in total phenolic contents. However, depending on processing time, significant increases or decreases of individual phenolic compounds were observed. PAW-10 and -20 increased the ascorbic acid content to 382.76 and 363.14 mg/100 g, respectively, compared to control (337.73 mg/100 g). Riboflavin and nicotinic acid contents were increased significantly in PAW-20 (0.53 and 1.26 mg/100), compared to control (0.32 and 0.61 mg/100 g, respectively). However, nicotinamide showed non-significant increase in all treatments. Antioxidant activity improved significantly only in PAW-20, while peroxidase activity was reduced up to 36% in the longest treatment. In conclusion, PAW treatment could be an effective technique for rocket decontamination since it positively influenced the quality of rocket, improving the retention of polyphenols and vitamins

Cementless ceramic-on-ceramic total hip replacement in children and adolescents

Background: total hip replacement (THR) is a rare surgical option in children and adolescents with disabling hip diseases. The aim of this study is to report results from a retrospective cohort of patients aged 18 years or less who underwent cementless Ceramic-on-Ceramic (CoC) THR at a single institution, investigating clinical and radiographic outcomes, survival rates, and reasons for revision of the implants. Materials and methods: we queried the Registry of Prosthetic Orthopedic Implants (RIPO) to identify all children and adolescents undergoing THR between 2000 and 2019 at a single Institution. Inclusion criteria were patients undergoing cementless CoC THR, aged less than 18 years at surgery, followed for at least 2 years. Sixty-eight patients (74 hips) matched all the inclusion criteria and were enrolled in the study. We assessed the clinical and radiographic outcomes, the rate of complications, the survival rate, and reasons for revision of the implants. Results: The mean follow-up was 6.6 ± 4.4 years (range 2–20). The most frequent reason for THR was post-traumatic or chemotherapy-induced avascular necrosis (38%). The overall survival rate of the cohort was 97.6% (95% CI: 84.9–99.7%) at 5 years of follow-up, 94.4% (95% CI: 79.8–98.6%) at 10 years and 15 years of follow-up. Two THR in two patients (2.7%) required revision. With the numbers available, Cox regression analysis could not detect any significant interaction between preoperative or intraoperative variables and implant survivorship (p-value 0.242 to 0.989).” The average HOOS was 85 ± 14.3 (range 30.6–100). Overall, 23 patients (48%) reported excellent HOOS scores (&gt;90 points), 21 patients (44%) reported acceptable HOOS scores (60–90 points) while 4 patients (8%) reported poor outcomes (&lt;60 points). Twenty-one patients (43%) were regularly involved into moderate-to high-intensity sport activities (UCLA ≥ 6). Conclusions: Cementless CoC THR is a successful procedure in children and teenagers, having demonstrated high implant survivorship and low rates of complications and failure. A meticulous preoperative planning and implant selection is mandatory, to avoid implant malposition, which is the main reason of failure and revision in these cases. Further studies are needed to assess the impact of the THR on the psychosocial wellbeing of teenagers, as well as risks and benefits and cost-effectiveness in comparison to the hip preserving surgical procedures

PREVENZIONE DEI DISTURBI DELLO SVILUPPO DEL BAMBINO

Abstract Introduction: The literature and current experience tend to doubt the usefulness of the psychomotor rehabilitation in the cases of pre-perinatal neurodamage. The characteristics of brain development undoubtedly indicate that the first year of life and actually the first months are crucial for the post-natal development and re-organization of structures. These periods are characterised by a greater plasticity and therefore they are theoretically the most suitable for rehabilitation. Aim: To investigate the efficacy of the early application (within the first three months of life) of a specific educational-rehabilitative methodology (the one applied at “A.R.C. i Nostri figli”) in children presenting with neurodamage and in children at risk. Method: Retrospective observational study on patients treated in the rehabilitation centre A.R.C. i Nostri Figli from 1995 to 2005. Results: The results show a statistically significant improvement in the autonomy of patients that start the therapy before 3 months of age if compared with those that start it after that age. Conclusions: These observations need to be confirmed on children that have been treated with different therapies. We think that a controlled perspective rather than retrospective experimentation is not ethical. So, for the time being, we can conclude that any effort should be done for each single patient in order to anticipate the rehabilitation intervention to the first months of age

Desarrollo de productos de arándanos con propiedades antioxidantes y probióticas

Se estudiaron alternativas de industrialización de arándanos mediante la obtención de productos con propiedades funcionales. Se elaboró jugo de arándanos (JA) despectinizando con 4mg de enzima LAFASE®-CLARIFICATION/100g arándanos y 8mg enzima LAFASE®-HE-GRAND-CRU/100g arándanos a 50°C durante 1,3h. En estas condiciones se maximizaron el rendimiento de extracción de jugo y la concentración de antocianinas totales (AT). Para estabilizar el JA se optimizó el baremo temperatura-tiempo de pasteurización en 77°C y 85s, lo que posibilitó la obtención de jugo de arándanos pasteurizado (JAP) con altas concentraciones de AT. Al JAP se incorporaron bacterias probióticas de Lactobacillus rhamnosus, obteniendo jugo de arándanos con probióticos (JACP). Las mejores condiciones de almacenamiento para el jugo de arándanos fueron como JAP almacenado a 0°C durante 300 días y como JACP a 0°C durante 30 días. El JAP se utilizó para impregnar rodajas de manzana y zucchini y potenciar sus propiedades funcionales. Las rodajas fueron deshidratadas a 40, 50 y 60°C para obtener snacks enriquecidos con antocianinas. El modelo de Page describió adecuadamente la cinética de deshidratación. A 60°C se logró la mayor retención de AT y el menor tiempo de deshidratación, lo que es de interés desde el punto de vista tecnológic

Effect of production process and high-pressure processing on viability of Salmonella spp. in traditional Italian dry-cured coppa

The aim of the study was to investigate the combined effect of the manufacturing process followed by HPP treatment on the inactivation of Salmonella spp. in artificially contaminated coppa samples, in order to verify the ability of the combined processes to achieve the objective of a 5-log reduction of Salmonella spp. needed for exportation to the U.S. Fresh anatomical cuts intended for coppa production were supplied by four different delicatessen factories located in Northern Italy. Raw meat underwent experimental contamination with Salmonella spp. using a mixture of 3 strains. Surface contamination of the fresh anatomical cuts was carried out by immersion into inoculum containing Salmonella spp. The conditions of the HPP treatment were: pressure 593 MPa, time 290 seconds, water treatment temperature 14\ub0C. Surface and deep samples were performed post contamination (T0), end of the cold phase (T1), end of process (Tend), and after HPP treatment (postHPP) and Salmonella spp. Enumerated. The results of this study show a significant reduction of Salmonella spp. all through the production process (P&lt;0.01) for all companies, followed by an additional reduction of bacterial counts due to HPP treatment (P&lt;0.01), both in superficial and deep contaminations (P&lt;0.01). The superficial overall reduction resulted of 1.58 to 5.04 log CFU/g during the production process. HPP treatment resulted in a significant (P&lt;0.01) superficial and deep decrease in Salmonella spp. enumeration varying from 0.61 to 4.01 log and from 1.49 to 4.13 log. According to the data presented in this study, only the combined approach of coppa manufacturing process followed by HPP treatment always led to a 5-log reduction of Salmonella spp. required by USDA/FSIS guidelines