Effect of pasteurization on in vitro \u3b1-glucosidase inhibitory activity of apple juice

The in vitro alpha-glucosidase inhibitory activity of raw, mildly (F-71.7(5) = 0.4 min, 5 Log reductions of Cryptosporidium parvum) and intensively (F-90(12) = 14.8 min, 2 Log reductions of Alicyclobacillus acidoterrestris) pasteurized apple juice was studied. Raw apple juice (23 mg(dw)/mL) caused 90% alpha-glucosidase inhibition. Analogous results were obtained for the mildly treated sample. The most intense treatment reduced by 35% the alpha-glucosidase inhibition. However, such a decrease was associated with an increase in the phenolic content, suggesting that alpha-glucosidase inhibition might not rely on these compounds, but depend on more complex mechanisms. Apple juice was combined with acarbose to investigate their interaction towards alpha-glucosidase inhibition. A synergistic behavior was observed for concentrations < 2 mg/mL. Increasing the concentration of the combined system (up to 9 mg/mL) produced an antagonistic effect, while a further increase (< 9 mg/mL) allowed approaching an addictive behavior

Reformulation and food combination as strategies to modulate glycaemia: the case of apple pomace containing biscuits administered with apple juice to healthy rats

Conventional (CB) and apple-pomace-reformulated (RB) biscuits were administered to healthy rats. Although the areas under curve (AUC) of glucose concentration were comparable between samples, differences in the glycaemic profile of CB and RB were observed. RB caused an initial steeper increase in glycaemia but a shift in the glycaemic peak from 45 to 60 min, as compared to CB. When CB or RB was ingested with apple juice (AJ) no differences were observed as compared to their ingestion with a soft drink (SD) simulating AJ sugar content, indicating that reformulation, more than the presence of AJ, was crucial in affecting the glycaemic response. Consumer acceptability towards reformulation was assessed through conjoint analysis, by simulating labels reporting information on reformulation. Consumers preferred information generally referring to the health-promoting effect (i.e. \u201clow sugar\u201d and "high fibre\u201d contents), despite directly relating to a specific disease (i.e. \u201csuitable for diabetics\u201d and \u201clow glycaemic index\u201d)

Phenolic content and potential bioactivity of apple juice as affected by thermal and ultrasound pasteurization

Thermal (T) and ultrasound (US) pasteurization processes were applied to apple juice and the phenolic compounds (TPC) were quantified before and after in vitro digestion by HPLC-DAD-ESI-MSn, with their bioaccessibility ascertained. Digested samples were analysed for their inhibitory capacity against α-glucosidase. Since some of the compounds exhibit fluorescence, both steady state and time-resolved fluorescence methods were used to investigate the binding to a blood transport protein, human serum albumin (HSA). It was found that processing induced an increase in the TPC content, which was more pronounced when US was applied. In contrast, digestion reduced the TPC content, evening out the overall effect. Still T and US pasteurized juices exhibited a higher quantity of TPC upon digestion as compared to the raw sample. No correlation was found between the TPC content and α-glucosidase inhibition, as the T and US pasteurized juices showed the highest and lowest inhibitory capacities against the enzyme, respectively. This is indicative that other compounds, such as those formed upon thermal treatment, may be involved in the antidiabetic effect of apple juice. The fluorescence study showed that binding occurred to HSA, at slightly different rates for different species present in the US treated extract. Considering energy consumption, US pasteurization is the most power consuming treatment despite its shorter duration. Overall, no univocal indication on the best pasteurization process can be gathered. Thus, it is necessary to define the desired target in order to drive technological interventions by a customized approach.</p

Cydonia oblonga Mill. Pulp Callus Inhibits Oxidative Stress and Inflammation in Injured Cells

The pharmacological activity of a callus extract from the pulp of Cydonia oblonga Mill., also known as quince, was investigated in murine macrophage (RAW 264.7) and human keratinocyte (HaCaT) cell lines. In particular, the anti-inflammatory activity of C. oblonga Mill. pulp callus extract was assessed in lipopolysaccharides (LPS)-treated RAW264.7 by the Griess test and in LPS-treated HaCaT human keratinocytes by examining the expression of genes involved in the inflammatory process, including nitric oxide synthase (iNOS), interleukin-6 (IL-6), interleukin-1 (IL-1 ), nuclear factor-kappa-B inhibitor alfa (ikB ), and intercellular adhesion molecule (ICAM). The antioxidant activity was evaluated by quantizing the reactive oxygen species (ROS) production in the hydrogen peroxide and tert-butyl hydroperoxide-injured HaCaT cell line. The obtained results indicate that C. oblonga callus from fruit pulp extract has anti-inflammatory and antioxidant activities, suggesting its possible application in delaying and preventing acute or chronic diseases associated with aging or in the treatment of wound dressing

Altered Motoneuron Properties Contribute to Motor Deficits in a Rabbit Hypoxia-Ischemia Model of Cerebral Palsy

Cerebral palsy (CP) is caused by a variety of factors attributed to early brain damage, resulting in permanently impaired motor control, marked by weakness and muscle stiffness. To find out if altered physiology of spinal motoneurons (MNs) could contribute to movement deficits, we performed whole-cell patch-clamp in neonatal rabbit spinal cord slices after developmental injury at 79% gestation. After preterm hypoxia-ischemia (HI), rabbits are born with motor deficits consistent with a spastic phenotype including hypertonia and hyperreflexia. There is a range in severity, thus kits are classified as severely affected, mildly affected, or unaffected based on modified Ashworth scores and other behavioral tests. At postnatal day (P)0–5, we recorded electrophysiological parameters of 40 MNs in transverse spinal cord slices using whole-cell patch-clamp. We found significant differences between groups (severe, mild, unaffected and sham control MNs). Severe HI MNs showed more sustained firing patterns, depolarized resting membrane potential, and fired action potentials at a higher frequency. These properties could contribute to muscle stiffness, a hallmark of spastic CP. Interestingly altered persistent inward currents (PICs) and morphology in severe HI MNs would dampen excitability (depolarized PIC onset and increased dendritic length). In summary, changes we observed in spinal MN physiology likely contribute to the severity of the phenotype, and therapeutic strategies for CP could target the excitability of spinal MNs

Estimation of self-sustained activity produced by persistent inward currents using firing rate profiles of multiple motor units in humans

Persistent inward calcium and sodium currents (IP) activated during motoneuron recruitment help synaptic inputs maintain self-sustained firing until de-recruitment. Here, we estimate the contribution of the IP to self-sustained firing in human motoneurons of varying recruitment threshold by measuring the difference in synaptic input needed to maintain minimal firing once the IP is fully activated compared with the larger synaptic input required to initiate firing prior to full IP activation. Synaptic input to ≈20 dorsiflexor motoneurons simultaneously recorded during ramp contractions was estimated from firing profiles of motor units decomposed from high-density surface-EMG. To avoid errors introduced when using high-threshold units firing in their nonlinear range, we developed methods where the lowest-threshold units firing linearly with force were used to construct a composite (control) firing rate profile to estimate synaptic input to the higher-threshold (test) units. The difference in the composite firing rate (synaptic input) at the time of test unit recruitment and de-recruitment (ΔF=Frecruit-Fde-recruit) was used to measure IP amplitude that sustained firing. Test units with recruitment thresholds 1-30% of maximum had similar ΔFs, which likely included both slow and fast motor units activated by small and large motoneurons, respectively. This suggests that the portion of the IP that sustains firing is similar across a wide range of motoneuron sizes. Higher-threshold units had more prolonged accelerations in firing rate at the onset of recruitment compared to lower-threshold units, likely reflecting IP activation closer to firing onset in the higher-threshold units, but well before firing onset in the lower-threshold units

Prunus spinosa extract loaded in biomimetic nanoparticles evokes in vitro anti-inflammatory and wound healing activities

Prunus spinosa fruits (PSF) contain different phenolic compounds showing antioxidant and anti-inflammatory activities. Innovative drug delivery systems such as biomimetic nanoparti-cles could improve the activity of PSF extract by promoting (i) the protection of payload into the lipidic bilayer, (ii) increased accumulation to the diseased tissue due to specific targeting properties, (iii) improved biocompatibility, (iv) low toxicity and increased bioavailability. Using membrane proteins extracted from human monocyte cell line THP-1 cells and a mixture of phospholipids, we formulated two types of PSF-extract-loaded biomimetic vesicles differing from each other for the presence of either 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or 1,2-dioleoyl-sn-glycero-3-phospho-(1\u2032-rac-glycerol) (DOPG). The biological activity of free extract (PSF), compared to both types of extract-loaded vesicles (PSF-DOPCs and PSF-DOPGs) and empty vesicles (DOPCs and DOPGs), was evaluated in vitro on HUVEC cells. PSF-DOPCs showed preferential incorporation of the extract. When enriched into the nanovesicles, the extract showed a significantly increased anti-inflammatory activity, and a pronounced wound-healing effect (with PSF-DOPCs more efficient than PSF-DOPG) compared to free PSF. This innovative drug delivery system, combining nutraceuti-cal active ingredients into a biomimetic formulation, represents a possible adjuvant therapy for the treatment of wound healing. This nanoplatform could be useful for the encapsulation/enrichment of other nutraceutical products with short stability and low bioavailability

Prunus spinosa Extract Loaded in Biomimetic Nanoparticles Evokes In Vitro Anti-Inflammatory and Wound Healing Activities

none14sìPrunus spinosa fruits (PSF) contain different phenolic compounds showing antioxidant and anti-inflammatory activities. Innovative drug delivery systems such as biomimetic nanoparticles could improve the activity of PSF extract by promoting (i) the protection of payload into the lipidic bilayer, (ii) increased accumulation to the diseased tissue due to specific targeting properties, (iii) improved biocompatibility, (iv) low toxicity and increased bioavailability. Using membrane proteins extracted from human monocyte cell line THP-1 cells and a mixture of phospholipids, we formulated two types of PSF-extract-loaded biomimetic vesicles differing from each other for the presence of either 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DOPG). The biological activity of free extract (PSF), compared to both types of extract-loaded vesicles (PSF-DOPCs and PSF-DOPGs) and empty vesicles (DOPCs and DOPGs), was evaluated in vitro on HUVEC cells. PSF-DOPCs showed preferential incorporation of the extract. When enriched into the nanovesicles, the extract showed a significantly increased anti-inflammatory activity, and a pronounced wound-healing effect (with PSF-DOPCs more efficient than PSF-DOPG) compared to free PSF. This innovative drug delivery system, combining nutraceutical active ingredients into a biomimetic formulation, represents a possible adjuvant therapy for the treatment of wound healing. This nanoplatform could be useful for the encapsulation/enrichment of other nutraceutical products with short stability and low bioavailability.openTiboni, Mattia; Coppari, Sofia; Casettari, Luca; Guescini, Michele; Colomba, Mariastella; Fraternale, Daniele; Gorassini, Andrea; Verardo, Giancarlo; Ramakrishna, Seeram; Guidi, Loretta; Di Giacomo, Barbara; Mari, Michele; Molinaro, Roberto; Albertini, Maria CristinaTiboni, Mattia; Coppari, Sofia; Casettari, Luca; Guescini, Michele; Colomba, Mariastella; Fraternale, Daniele; Gorassini, Andrea; Verardo, Giancarlo; Ramakrishna, Seeram; Guidi, Loretta; Di Giacomo, Barbara; Mari, Michele; Molinaro, Roberto; Albertini, Maria Cristin