Disclosing the Antioxidant and Neuroprotective Activity of an Anthocyanin-Rich Extract from Sweet Cherry (Prunus avium L.) Using In Vitro and In Vivo Models

In this study, an autochthonous variety of sweet cherry (Prunus avium L.), namely “Moretta di Vignola”, was processed to prepare extracts rich in polyphenols, which were characterized by high-performance liquid chromatography (HPLC) separation coupled to UV/DAD and ESI-MSn analysis. Then, a sweet cherry anthocyanin-rich extract (ACE) was prepared, fully characterized and tested for its activity against Parkinson’s disease (PD) in cellular (BV2 microglia and SH-SY5Y neuroblastoma) and in Drosophila melanogaster rotenone (ROT)-induced model. The extract was also evaluated for its antioxidant activity on Caenorhabditis elegans by assessing nematode resistance to thermal stress. In both cell lines, ACE reduced ROT-induced cell death and it decreased, alone, cellular reactive oxygen species (ROS) content while reinstating control-like ROS values after ROT-induced ROS rise, albeit at different concentrations of both compounds. Moreover, ACE mitigated SH-SY5Y cell cytotoxicity in a non-contact co-culture assay with cell-free supernatants from ROT-treated BV-2 cells. ACE, at 50 µg/mL, ameliorated ROT (250 µM)-provoked spontaneous (24 h duration) and induced (after 3 and 7 days) locomotor activity impairment in D. melanogaster and it also increased survival and counteracted the decrease in fly lifespan registered after exposure to the ROT. Moreover, heads from flies treated with ACE showed a non-significant decrease in ROS levels, while those exposed to ROT markedly increased ROS levels if compared to controls. ACE + ROT significantly placed the ROS content to intermediate values between those of controls and ROT alone. Finally, ACE at 25 µg/mL produced a significant increase in the survival rate of nematodes submitted to thermal stress (35 °C, 6–8 h), at the 2nd and 9th day of adulthood. All in all, ACE from Moretta cherries can be an attractive candidate to formulate a nutraceutical product to be used for the prevention of oxidative stress-induced disorders and related neurodegenerative diseases

CCAP regulates feeding behavior via the NPF pathway in Drosophila adults

The intake of macronutrients is crucial for the fitness of any animal and is mainly regulated by peripheral signals to the brain. How the brain receives and translates these peripheral signals or how these interactions lead to changes in feeding behavior is not well-understood. We discovered that 2 crustacean cardioactive peptide (CCAP)-expressing neurons in Drosophila adults regulate feeding behavior and metabolism. Notably, loss of CCAP, or knocking down the CCAP receptor (CCAP-R) in 2 dorsal median neurons, inhibits the release of neuropeptide F (NPF), which regulates feeding behavior. Furthermore, under starvation conditions, flies normally have an increased sensitivity to sugar; however, loss of CCAP, or CCAP-R in 2 dorsal median NPF neurons, inhibited sugar sensitivity in satiated and starved flies. Separate from its regulation of NPF signaling, the CCAP peptide also regulates triglyceride levels. Additionally, genetic and optogenetic studies demonstrate that CCAP signaling is necessary and sufficient to stimulate a reflexive feeding behavior, the proboscis extension reflex (PER), elicited when external food cues are interpreted as palatable. Dopaminergic signaling was also sufficient to induce a PER. On the other hand, although necessary, NPF neurons were not able to induce a PER. These data illustrate that the CCAP peptide is a central regulator of feeding behavior and metabolism in adult flies, and that NPF neurons have an important regulatory role within this system

EFFECT OF THE D2-AUTORECEPTOR AGONIST B-HT-958 ON BOTH SPONTANEOUS AND ACTH-INDUCED STRETCHING, YAWNING AND GROOMING IN THE RAT

The D2 autoreceptor agonist B-HT 958, intraperitoneally injected into Wistar male rats in a novel environment, significantly increased stretching and yawning (SY) while inhibiting grooming. Pretreatment with the D2 antagonist sulpiride reversed these effects, antagonizing SY and restoring grooming. Similarly, when B-HT 958 was administered to rats in their home cages, it elicited SY and abolished grooming; moreover, when administered before the i.c.v. injection of adrenocorticotropin hormone, dose-dependently enhanced SY and strongly antagonized the typical syndrome of intensified grooming induced by the peptide. The possible relationship between SY and grooming and the involvement of D2 autoreceptors are discussed

Single center experience with the Sorin Bicarbon prosthesis. A 17-year follow-up

Objective: To evaluate the long-term results of aortic valve replacement (AVR) and mitral valve replacement (MVR) with the Sorin Bicarbon prosthesis (SBP). Methods: Five hundred seven patients (306 men, 201 women), mean age 62 10 years (range, 21-86 years), received an SBP between 1994 and 2000; AVR was performed in 344 (67%) and MVR in 163 (33%). The main concomitant procedure was coronary artery grafting in 79 patients (16%). Follow-up was 99%complete; mean follow-up was 12.7 4.0 years with a cumulative duration of follow-up of 6475 patient-years in the entire group (4348 patient-years for AVR and 2124 patient-years for MVR). Results: Hospital mortality was 2.7% (AVR, 2.03%; MVR, 4.3%). There were 169 late deaths (AVR, 128; MVR, 41). Actuarial survival at 17 years is 49.7% 5.3% for AVR and 62.0% 6.1% for MVR. At the last follow-up, 310 survivors (199 AVR, 111 MVR) are in New York Heart Association functional class I or II. At 17 years, actuarial freedom from valve-related deaths, embolism, and bleeding is 89.8% 4.8%, 85.8% 5.4%, and 96.2% 1.2% after AVR, and 91.9% 3.9%, 96.3% 1.8%, 95.0% 2.9% after MVR. Reoperation was required in 5 patients with AVR (thrombosis in 4 and perivalvular leak in 1). Actuarial freedom from reoperation is 98.1% 0.8% after AVR and 100% after MVR; freedom from endocarditis is 100% after AVR and 99.2% 0.7% after MVR. No cases of intrinsic structural valve failure were observed. Conclusions: The SBP has shown excellent results in terms of clinical improvement and freedom from valve-related complications, even up to 17 years after AVR and MVR. It therefore seems to be a safe option whenever a mechanical prosthesis is needed. (J Thorac Cardiovasc Surg 2014;148:2039-44

Influence of SAMe on the modifications of brain poliamine levels in an animal model of depression

The mechanism(s) of the antidepressant activity of S-adenosyl-L-methionine (SAMe) have not yet been elucidated. SAMe is essential for the synthesis of polyamines, which have a key role in protein synthesis, cell proliferation, and neuronal plasticity. On the other hand, accumulating data indicate that depression is associated with a reduction in regional brain volume and that antidepressants increase neurogenesis in defined brain regions and also influence neuronal plasticity. Here we show that in a validated rat model of depression (chronic unpredictable mild stress-induced anhedonia) there is a significant reduction of putrescine, spermidine and spermine in the hippocampus, and of only putrescine in the nucleus accumbens septi. SAMe, at a fully antidepressant dose (300 mg/kg i.m., daily for 7 days), completely restores the levels of putrescine in the nucleus accumbens, and restores in part the levels of both spermidine and spermine in the hippocampus. These results may suggest (i) a role for brain polyamines in depression and in reward processes, and (ii) that the antidepressant effect of SAMe may be due, at least in part, to a normalization of putrescine levels in the nucleus accumbens septi

Influence of UFP-101 central infusion on behavioural and cellular effects in a chronic stress model in the rat.

Nociceptin/orphanin FQ (N/OFQ) and its receptor (NOP) modulate several functions in the central nervous system. UFP-101 is a selective and high affinity NOP receptor antagonist [1], that has been reported to elicit antidepressant-like effects in rodents. The present study investigated the effect of UFP-101 in the chronic mild stress (CMS) paradigm in male Wistar rats. Animals were exposed to CMS for at least seven weeks, to induce a condition of anhedonia (measured as reduced consumption of a sucrose solution). UFP-101 (10 nmol/rat, i.c.v. continuously infused by means of minipumps for 24 days) did not influence sucrose intake in non-stressed animals, but reinstated basal sucrose consumption in stressed animals (+ 40% vs non treated stressed rats), beginning from the second week of treatment. The reference drug fluoxetine (FL, 10 mg/kg, i.p.) produced identical effects. Similarly, in the forced swimming test (FST) performed 2, 9, 16 and 23 days after treatment, UFP-101 reverted the effects of stress in a dose- and time-dependent manner, reducing the time of immobility of stressed rats to the level of non-stressed controls (−78% vs non treated stressed rats); FL produced the same effect from day 9 onwards. Moreover, repeated co-administration of N/OFQ (5 nmol/rat i.c.v., from day 15 to 24) completely prevented the behavioural effects of UFP-101. In previous animal studies the stress/depression condition has been associated to plastic alterations of neuronal networks, including reduced hippocampal volume, neuronal atrophy, cell death and alterations in cell proliferation and cell survival of newly generated neurons in key limbic brain region implicated in depression [2], [3]. Neurotrophic factors (NTFs) such as BDNF and FGF-2 may be involved in the modulation of activitydependent plasticity associated with mood pathologies. Therefore, it was investigated whether the CMS procedure associated with continuous central infusion of UFP-101 can affect NTFs expression, as well as proliferation and survival of hippocampal newborn cells. After in vivo BrdU labelling, all rats were sacrificed on day 24 and their brains were collected for immunohistochemical analysis. Our preliminary data confirm that CMS reduces proliferation of neural stem cells in hippocampus. The 21-day UFP-101 treatment, which did not produce any primary effect on cell proliferation, also did not affect the reduced proliferation observed in stressed animals. Experiments are now in progress to evaluate the NTFs expression levels in discrete brain areas. These findings support the view that blockade of NOP receptor signaling in the brain produces antidepressantlike effects in the CMS protocol, and indicate that the behavioural efficacy of the NOP receptor antagonist UFP- 101 is comparable to that of classical antidepressants. However, this effect does not appear to be mediated by increased hippocampal stem cell proliferation. These findings support the hypothesis that NOP receptor may represent a candidate target for innovative antidepressant drugs

Single center experience with the Sorin Bicarbon prosthesis: A\ua017-year clinical follow-up.

To evaluate the long-term results of aortic valve replacement (AVR) and mitral valve replacement (MVR) with the Sorin Bicarbon prosthesis (SBP).Five hundred seven patients (306 men, 201 women), mean age 62 \ub1 10 years (range, 21-86 years), received an SBP between 1994 and 2000; AVR was performed in 344 (67\%) and MVR in 163 (33\%). The main concomitant procedure was coronary artery grafting in 79 patients (16\%). Follow-up was 99\% complete; mean follow-up was 12.7 \ub1 4.0 years with a cumulative duration of follow-up of 6475 patient-years in the entire group (4348 patient-years for AVR and 2124 patient-years for MVR).Hospital mortality was 2.7\% (AVR, 2.03\%; MVR, 4.3\%). There were 169 late deaths (AVR, 128; MVR, 41). Actuarial survival at 17 years is 49.7\% \ub1 5.3\% for AVR and 62.0\% \ub1 6.1\% for MVR. At the last follow-up, 310 survivors (199 AVR, 111 MVR) are in New York Heart Association functional class I or II. At 17 years, actuarial freedom from valve-related deaths, embolism, and bleeding is 89.8\% \ub1 4.8\%, 85.8\% \ub1 5.4\%, and 96.2\% \ub1 1.2\% after AVR, and 91.9\% \ub1 3.9\%, 96.3\% \ub1 1.8\%, 95.0\% \ub1 2.9\% after MVR. Reoperation was required in 5 patients with AVR (thrombosis in 4 and perivalvular leak in 1). Actuarial freedom from reoperation is 98.1\% \ub1 0.8\% after AVR and 100\% after MVR; freedom from endocarditis is 100\% after AVR and 99.2\% \ub1 0.7\% after MVR. No cases of intrinsic structural valve failure were observed.The SBP has shown excellent results in terms of clinical improvement and freedom from valve-related complications, even up to 17 years after AVR and MVR. It therefore seems to be a safe option whenever a mechanical prosthesis is needed

Single center experience with the Sorin Bicarbon prosthesis: A 17-year clinical follow-up

To evaluate the long-term results of aortic valve replacement (AVR) and mitral valve replacement (MVR) with the Sorin Bicarbon prosthesis (SBP).Five hundred seven patients (306 men, 201 women), mean age 62 ± 10 years (range, 21-86 years), received an SBP between 1994 and 2000; AVR was performed in 344 (67\%) and MVR in 163 (33\%). The main concomitant procedure was coronary artery grafting in 79 patients (16\%). Follow-up was 99\% complete; mean follow-up was 12.7 ± 4.0 years with a cumulative duration of follow-up of 6475 patient-years in the entire group (4348 patient-years for AVR and 2124 patient-years for MVR).Hospital mortality was 2.7\% (AVR, 2.03\%; MVR, 4.3\%). There were 169 late deaths (AVR, 128; MVR, 41). Actuarial survival at 17 years is 49.7\% ± 5.3\% for AVR and 62.0\% ± 6.1\% for MVR. At the last follow-up, 310 survivors (199 AVR, 111 MVR) are in New York Heart Association functional class I or II. At 17 years, actuarial freedom from valve-related deaths, embolism, and bleeding is 89.8\% ± 4.8\%, 85.8\% ± 5.4\%, and 96.2\% ± 1.2\% after AVR, and 91.9\% ± 3.9\%, 96.3\% ± 1.8\%, 95.0\% ± 2.9\% after MVR. Reoperation was required in 5 patients with AVR (thrombosis in 4 and perivalvular leak in 1). Actuarial freedom from reoperation is 98.1\% ± 0.8\% after AVR and 100\% after MVR; freedom from endocarditis is 100\% after AVR and 99.2\% ± 0.7\% after MVR. No cases of intrinsic structural valve failure were observed.The SBP has shown excellent results in terms of clinical improvement and freedom from valve-related complications, even up to 17 years after AVR and MVR. It therefore seems to be a safe option whenever a mechanical prosthesis is needed

Differential Sensitivity of A(2A) and Especially D(2) Receptor Trafficking to Cocaine Compared with Lipid Rafts in Cotransfected CHO Cell Lines. Novel Actions of Cocaine Independent of the DA Transporter

The effects of low and high concentrations of cocaine have been studied in vitro on the trafficking of plasma membrane A(2A) and D(2) immunoreactivities in previously characterized A(2A)-D(2) CHO cell lines. Receptor double immunofluorescence staining was performed with D(2) and A(2A) antibodies, planar lipid rafts immunolabeling with biotinylated cholera toxin subunit B and membrane invaginations with an anti-caveolin-1 antibody. A computer-assisted image analysis demonstrated a substantial and highly significant rise of membrane-associated D(2) immunoreactivity (IR) after 8 h of exposure to a low concentration of cocaine (150 nM). At this low concentration of cocaine, there was also an increase of membrane associated A(2A) immunoreactivity but smaller and less significant. However, this increase became considerably larger and highly significant at 150 microM at which concentration the rise of D(2) immunoreactivity had begun to disappear. It may be suggested that an allosteric action of cocaine at 150 nM on the D(2) receptors may primarily increase the insertion of D(2) monomers, homomers and also of a subpopulation of A(2A)-D(2) heteromers from the cytoplasm into the plasma membrane due to the conformational change induced by cocaine in the D(2) receptor. The planar lipid rafts and the caveolae are only affected by the higher concentrations of cocaine. It is proposed that changes in D(2) and A(2A)-D(2) trafficking induced by allosteric actions of cocaine at D(2) receptors may contribute to the alterations of D(2) signaling found in cocaine abusers

Anxiolytic-like effect of neuropeptide S in the rat defensive burying.

Neuropeptide S (NPS) has been recently identified as the endogenous ligand of a previously orphan G-protein-coupled receptor now named NPSR. Both NPS and its receptor are expressed in the brain, where they modulate different functions. In particular, it has been demonstrated that intracerebroventricular (i.c.v.) injection of NPS in rodents increases wakefulness and promotes anxiolytic-like effects. In the present study we used the defensive burying (DB) test in rats to further investigate the action of human NPS (0.1-10nmol, i.c.v.) on anxiety-related behaviors. Diazepam (1.5mg/kg, i.p.) and caffeine (20mg/kg, i.p.) were used in parallel experiments as standard anxiolytic and anxiogenic drugs, respectively. None of the tested drugs produced statistical differences in the latency to contact the probe, burying behavior latency, number of shocks received or immobility/freezing duration. Caffeine increased cumulative burying behavior and the buried bedding height in a statistically significant manner thus promoting anxiogenic like effects. Opposite results were obtained with diazepam that significantly reduced these behavioral parameters. The anxiolytic-like action of diazepam was mimicked by NPS that reduced cumulative burying behavior in a dose dependent manner. Collectively, robust anxiolytic-like effects were recorded in response to NPS in the DB test. These results are of particular interest since the outcome of this assay is marginally influenced by drug effects on locomotor activity. In conclusion, we provide further evidence that NPS evokes genuine anxiolytic-like effects in the rat; therefore NPSR selective agonists are worthy of development as innovative drugs for the treatment of anxiety disorders