Prickly Pear Betalain-Rich Extracts as New Promising Strategy for Intestinal Inflammation: Plant Complex vs. Main Isolated Bioactive Compounds.

Recently, many studies have highlighted the health effects of betalains beyond their use as food dyes. The present study investigated betalain-rich extracts with different colors and their main bioactive compounds in order to provide first evidence as a new promising strategy for intestinal inflammation management. Prickly pear betalain-rich extracts, obtained by a QuEChERS method, have been characterized by LC-DAD-ESI-MS/MS analysis. The potential role of betanin, indicaxanthin, and prickly pear extracts in counteracting the antioxidant and anti-inflammatory events was evaluated by several in vitro cell-free and cell-based assays. Indicaxanthin and betanin represent the most abundant compounds (≥22.27 ± 4.50 and 1.16 ± 0.17 g/100 g dry extract, respectively). Prickly pear extracts showed the strongest antioxidant and anti-inflammatory activities with respect to the pure betalains both on in vitro cell-free and cell-based assays, demonstrating the occurrence of synergistic activity, without any cytotoxicity or alteration of the barrier systems. The release of reactive oxygen species (ROS) and key inflammatory markers (IL-6, IL-8, and NO) was strongly inhibited by both betalains and even more by prickly pear extracts, which showed a similar and sometimes better profile than the reference compounds trolox and dexamethasone in counteracting the IL-1ß-induced intestinal inflammation.

Massive infection of Cystidicoloides ephemeridarum in brown trout Salmo trutta with skeletal deformities.

We investigated the cause of skeletal deformities found in brown trout from the Aspromonte mountain area in Reggio Calabria, Italy. Toxicological, histopathological and parasitological analyses were carried out on 14 fish with evident macro-morphological alterations from 2 different locations in the same river, and 4 control fish without morphological alterations from a different river (far from the first river but still within the area under study). Histopathological and radiological observations confirmed severe skeletal deformities in the specimens investigated. Parasitological examinations highlighted the presence of the nematode Cystidicoloides ephemeridarum, found only within the gastrointestinal tract of specimens showing deformities. Moreover, a direct correlation between parasite number and fish size was found. Given the low heavy metal levels and the presence of a massive parasitosis in teleosts showing deformities, we postulate a correlation between skeletal deformities and nematode infestation: the parasites caused a serious vitamin and mineral deficiency in the fish, which led to a dysplastic vertebral column. The low calcium levels found in malformed specimens compared with negative controls effectively confirm this hypothesis.

The problem of misidentification between edible and poisonous wild plants: Reports from the Mediterranean area.

Today, in many European countries, people are looking for wild edible plants to experience new tastes and flavors, by following the new trend of being green and environmentally friendly. Young borage and spinach leaves can be easily confused by inexpert pickers with those of other plants, including poisonous ones, such as Mandragora autumnalis Bertol. (mandrake) or Digitalis purpurea L. (foxglove), common in southern and northern Italy respectively. In the last twenty years, several cases of intoxication by accidental ingestion of mandrake and foxglove have been reported. The purpose of this work was to perform a pharmacognostic characterization of young leaves from borage, mandrake, foxglove and spinach, by micro-morphological, molecular and phytochemical techniques. The results showed that each of the three techniques investigated could be sufficient alone to provide useful information for the identification of poisonous species helping the medical staff to manage quickly the poisoned patients. However, the multi-disciplinary approach proposed could be very useful to asses the presence of poisonous plants in complex matrices, to build a database containing morphological, molecular and phytochemical data for the identification of poisonous species or in forensic toxicology, given their increasingly frequent use due to their low cost and relatively common availability.

Epigenetic versus Genetic Deregulation of the KEAP1/NRF2 Axis in Solid Tumors: Focus on Methylation and Noncoding RNAs.

Oxidative and electrophilic changes in cells are mainly coordinated by the KEAP1/NRF2 (Kelch-like erythroid-derived cap-n-collar homology- (ECH-) associated protein-1/nuclear factor (erythroid-derived 2)-like 2) axis. The physical interaction between these two proteins promotes the expression of several antioxidant defense genes in response to exogenous and endogenous insults. Recent studies demonstrated that KEAP1/NRF2 axis dysfunction is also strongly related to tumor progression and chemo- and radiotherapy resistance of cancer cells. In solid tumors, the KEAP1/NRF2 system is constitutively activated by the loss of KEAP1 or gain of NFE2L2 functions that leads to its nuclear accumulation and enhances the transcription of many cytoprotective genes. In addition to point mutations, epigenetic abnormalities, as aberrant promoter methylation, and microRNA (miRNA) and long noncoding RNA (lncRNA) deregulation were reported as emerging mechanisms of KEAP1/NRF2 axis modulation. This review will summarize the current knowledge about the epigenetic mechanisms that deregulate the KEAP1/NRF2 cascade in solid tumors and their potential usefulness as prognostic and predictive molecular markers.

Experimental exposure of blue mussels (Mytilus galloprovincialis) to high levels of benzo[a]pyrene and possible implications for human health.

Polycyclic aromatic hydrocarbons (PAHs) are lipophilic compounds able to accumulate in the food chain. Mussels showed to bioaccumulate contaminants, such as PAHs, so that recurrent consumption of such contaminated food represents a risk for human health. This study was aimed to elucidate if acute exposure of Mediterranean blue mussel (Mytilus galloprovincialis), a bivalve of great economic importance in several countries, to a PAH, benzo[a]pyrene (B[a]P), at doses able to induce cytochrome P450 1A (CYP1A) and pathological changes in mussel gills, can produce accumulation in soft tissue. We explored the cytotoxic effects (cell viability, DNA laddering, and glutathione levels) of in vitro exposure of human peripheral blood mononuclear cells (PBMCs) to organic extracts obtained from blue mussels previously exposed for 12 and 72h via water to B[a]P (0.5-1mg/L). In our experimental conditions, B[a]P induced CYP1A induction and morphological changes in mussel gills and a significant B[a]P accumulation in soft tissue. Conversely, exposing PBMCs to organic extracts obtained from contaminated mussels, resulted in a significant reduction of cell viability and cell glutathione content, and in an increase in DNA laddering. This confirms that consumption of mussels from B[a]P polluted waters might affect human health. Our data lead us to suggest that CYP1A activity in mussel gills may be useful (more than the amount of detected PAHs in the mussel edible tissue) as a marker in assessment of risk for health of consumers exposed to PAHs through ingestion of shellfish.

Polyphenolic profile and biological activities of black carrot crude extract (Daucus carota L. ssp. sativus var. atrorubens Alef.).

Black carrot (Daucus carota L. ssp. sativus var. atrorubens Alef.) is a valuable source of carbohydrates, minerals and vitamins and contains also high amounts of anthocyanins giving the characteristic deep-purple color. These latter compounds are known as natural dyes used in the food and pharmaceutical industry that have recently attracted much attention for their healthful properties. The aim of this work was to investigate for the first time the polyphenolic profile and biological properties of a black carrot crude extract (BCCE) through an in-depth analysis of the main polyphenolic classes evaluating its antioxidant, cytoprotective and anti-angiogenic properties. Twenty five polyphenols were quantified by LC-DAD-FLD-MS/MS analysis (anthocyanins 78.06%, phenolic acids 17.89% and other flavonoids 4.06%) with polyglycosylated cyanidins as major components. In addition, BCCE showed a strong antioxidant and free radical scavenging activity particularly in the hydrogen transfer-based assays (ORAC and ß-carotene bleaching) and a significant increase in the cell viability. Furthermore, BCCE exhibited a strong anti-angiogenic activity at the highest concentration assayed on the chick chorioallantoic membrane (50µg/egg). In conclusion, the obtained results demonstrated the antioxidant, cytoprotective and anti-angiogenic properties of BCCE, which highlight that the higher biological activity of BCCE is probably due to the synergic effects exerted by various polyphenolic classes.

Protective activity of an anthocyanin-rich extract from bilberries and blackcurrants on acute acetaminophen-induced hepatotoxicity in rats.

Acetaminophen (N-acetyl-p-aminophenol, APAP) overdosage can produce fatal centrilobular hepatic necrosis in humans. The present study attempted to investigate the protective effect of an anthocyanin-rich extract from bilberries and blackcurrants (AE) against APAP-induced acute hepatic damage in rats. Treatment with AE normalised blood activities of glutamate oxaloacetate and glutamate pyruvate transaminase and prevented APAP-induced plasmatic and tissutal alterations in biomarkers of oxidative stress, probably due to various bioproperties of the components of the extract.

Exposure of sea bream (Sparus aurata) to toxic concentrations of benzo[a]pyrene: possible human health effect.

Polycyclic aromatic hydrocarbons (PAHs) can accumulate in the food chain, due to their lipophilic properties. Fish can accumulate contaminants including PAHs and frequent consumption of such contaminated fish can pose risk to human health. The aim of this study was to clarify if acute exposure of sea bream (Sparus aurata, a fish species of great economic importance in the Atlantic and Mediterranean areas) to a PAH, benzo[a]pyrene (B[a]P), at a dose that can induce CYP1A and pathological changes in fish gills, liver and muscle, can induce accumulation in muscle. We investigated the cytotoxic effects (as changes in cell viability, DNA laddering and glutathione content) of in vitro exposure of human peripheral blood mononuclear cells (PBMCs) to organic extracts obtained from muscle of sea breams previously exposed via water to B[a]P (2mg/l, for 12, 24 and 72 h). At this level of exposure, B[a]P caused morphological changes, inflammatory response and CYP1A induction not only in sea bream gills and liver but also in muscle; furthermore, in fish muscle we observed a substantial B[a]P accumulation, which may be associated with the increased CYP1A activity in liver and especially in muscle. However, when PBMCs were exposed to organic extracts obtained from sea bream muscle contaminated with B[a]P, a toxic, although modest effect was revealed, consisting in a significant decrease in cell glutathione levels without alterations in cell viability and DNA laddering. This suggests that consumption of sea breams from B[a]P contaminated waters might represent a risk for human health.

Pharmacokinetics and efficacy of teicoplanin against intramammary infections in sheep.

The pharmacokinetics of teicoplanin were evaluated in 10 healthy sheep. After intravenous injection it showed monophasic behaviour, with a short mean (sd) elimination half-life (5 [0.24] hours). After intramuscular injection its bioavailability was 100 per cent but it was absorbed slowly; its elimination half-life was satisfactorily slow (9.23 [0.74] hours). Its efficacy was evaluated in 26 crossbreed sheep with milk positive to strains of Staphylococcus aureus, coagulase-negative staphylococci and Streptococcus agalactiae, and clinical signs of disease. In 19 of them a single intramuscular dose of 6 mg/kg bodyweight resulted in a complete resolution of the clinical signs, and no microorganisms were detected in milk by the fifth day; in the other seven sheep, with more severe intramammary infections, three consecutive daily intramuscular doses of 6 mg/kg bodyweight were necessary. There were no local or systemic side effects and no relapses during 30 days after the treatments in any of the sheep.

Common buzzards (Buteo buteo) bio-indicators of heavy metals pollution in Sicily (Italy).

The aim of this study was to evaluate the accumulation of toxic (As, Cd and Pb) and essential (Cu, Mn and Zn) metals in samples (feathers, liver, kidney, lung, intestine and muscle) of common buzzards (Buteo buteo) from Sicily, used as bio-indicators for monitoring environmental metals pollution. All samples of buzzards were collected at the "Recovery Center of Wild Fauna" of Palermo, through the Zooprophilactic Institute. The quantitative determinations of metals were carried out using an atomic absorbtion spectrophotometer (AAS). The results obtained showed the presence of metals in all samples analyzed. For toxic metals the highest levels of Pb and As were found in liver and those of Cd in kidney; for essential metals Zn levels were higher than Cu and Mn in all tissues analyzed. Significant differences are observed in metal levels between female and male and juvenile and adult bird samples. Highest metal levels found in liver, kidney and muscle can be considered indicative of chronic exposure to metals while the presence of metals in feathers can be consequential to storing and elimination processes. The results obtained suggest that common buzzards (Buteo buteo) may be very useful as bio-indicators for monitoring environmental pollution.

Presence of heterocyclic aromatic amines (HAS) in smoked "Provola" cheese from Calabria (Italy).

This study evaluated the residual levels of heterocyclic aromatic amines (HAs) in "Provola" cheese samples from Calabria, smoked using natural methods and using commercial buffered smoke. A comparative study of HAs concentrations was carried out on different portions of these samples: the rind, exterior part, core and on slice. Quantitative determination of HAs was carried out by HPLC using a fluorescence detector and analysis in HPLC-MS was conduced to confirm the presence of these amines. Residual levels of HAs were found in all naturally smoked "Provola" cheese samples. The results obtained showed that the smoking process performed using traditional methods contributes to HAs contamination while the use of commercial buffered smoke can be considered a safer technique for smoking of food.

Electron beam irradiated UHMWPE: degrading action of air and hyaluronic acid.

Ultra high molecular weight polyethylene, both pure and electron beam irradiated, was exposed to air and hyaluronic acid. Physical and mechanical analyses were performed in order to check the polymeric change due to the treatment. Pure UHMWPE, in fact, was modified by the hyaluronic acid that plasticizes the polymer. The electron irradiation cross links the chains and prevents their sliding in acid presence. A low irradiation dose is preferable rather than an high one. Low doses induce low amount of free radicals, witch react with oxygen and hyaluronic acid inducing low polymer degrade maintaining the initial mechanical performance. High doses degrade significantly the polymer properties.100 kGy irradiation in air can be applicable on the polyethylene as well as a compromise between the low mechanical performance improving and the material degradation.

Antiviral and immunomodulatory effect of a lyophilized extract of Capparis spinosa L. buds.

Herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) are common human pathogens that in particular cases can also cause severe problems especially in immunodeficient patients. The present paper reports the antiviral and immunomodulatory properties of a methanolic extract of C. spinosa buds (CAP), rich in flavonoids, including several quercetin and kaempferol glycosides. In particular we have investigated whether the in vitro exposure of human peripheral blood mononuclear cells (PBMCs) to CAP might inhibit the replication of HSV-2 and modulate the induction kinetics of IL-12, TNF-alpha IFN-gamma. Our findings have shown that CAP treatment interferes with HSV-2 replication in PBMCs inhibiting the extracellular virus release upregulating their production of IL-12, IFN-gamma and TNF-alpha. One could speculate that CAP may contribute in improving immune surveillance of PBMCs toward virus infection by up-regulating expression of peculiar proinflammatory cytokines; it should thus be successfully employed for treatment of HSV-2 infections in immunocompromised hosts.

Antimicrobial activity of flavonoids extracted from bergamot (Citrus bergamia Risso) peel, a byproduct of the essential oil industry.

AIMS: To evaluate the antimicrobial properties of flavonoid-rich fractions derived from bergamot peel, a byproduct from the Citrus fruit processing industry and the influence of enzymatic deglycosylation on their activity against different bacteria and yeast. METHODS AND RESULTS: Bergamot ethanolic fractions were tested against Gram-negative bacteria (Escherichia coli, Pseudomonas putida, Salmonella enterica), Gram-positive bacteria (Listeria innocua, Bacillus subtilis, Staphylococcus aureus, Lactococcus lactis) and the yeast Saccharomyces cerevisiae. Bergamot fractions were found to be active against all the Gram-negative bacteria tested, and their antimicrobial potency increased after enzymatic deglycosylation. The minimum inhibitory concentrations of the fractions and the pure flavonoids, neohesperidin, hesperetin (aglycone), neoeriocitrin, eriodictyol (aglycone), naringin and naringenin (aglycone), were found to be in the range 200 to 800 microg ml(-1). The interactions between three bergamot flavonoids were also evaluated. CONCLUSION: The enzyme preparation Pectinase 62L efficiently converted common glycosides into their aglycones from bergamot extracts, and this deglycosylation increased the antimicrobial potency of Citrus flavonoids. Pairwise combinations of eriodictyol, naringenin and hesperetin showed both synergistic and indifferent interactions that were dependent on the test indicator organism. SIGNIFICANCE AND IMPACT OF THE STUDY: Bergamot peel is a potential source of natural antimicrobials that are active against Gram-negative bacteria.

Effect of ozonation on sludge reduction in a SBR plant.

This paper provides new insights on the application of the ozonation process for the reduction of activated sludge production in a Sequencing Batch Reactor. The study was performed on two identical lab-scale SBRs plant, fed with domestic sewage: a fraction (1/3 of the working volume) of the activated sludge from one reactor (Exp SBR) was periodically subjected to ozonation for 30 minutes at 0.05 g O(3)/gSS and then recirculated before the beginning of the cycle; the other reactor was used as control and therefore managed at the same sludge retention time but without the application of ozonation. The effects of the recirculation of the ozonated sludge to the Exp SBR were evaluated in terms of biological nitrogen and carbon removal efficiencies, Mixed Liquor Volatile and Suspended Solids (MLSS and MLVSS, respectively) concentrations, effluent quality and sludge settleability. Besides, characterization of the ozonated sludge was carried out for different oxidant dosages (0.05, 0.07 and 0.37 g O(3)/gSS) and durations of the ozonation process (10, 20 and 30 minutes). The results show that at 0.05 g O(3)/gSS and 30 minutes contact time MLVSS as well as MLVSS/MLSS ratio do not change appreciably. Ozone dosage must be increased much further to obtain a relevant effect.

In vitro protective effect of a Jacquez grapes wine extract on UVB-induced skin damage.

Several studies have shown that UV radiation on the skin results in the formation of reactive oxygen species (ROS) that interact with proteins, lipids and DNA, thus altering cellular functions. The epidermis is composed mainly of keratinocytes, rich in ROS detoxifying enzymes and in low-molecular-mass antioxidant molecules. However, the increased generation of ROS can overwhelm the natural defences against oxidative stress. Therefore treatment of the skin with products containing plant-derived antioxidant ingredients may be a useful strategy for the prevention of UV-mediated cutaneous damage. In the present study we have investigated the in vitro capability of a Jacquez grapes wine extract (containing a significant level of proanthocyanidins, together with lower amounts of anthocyanins and hydroxycinnamic acids; JW-E), to protect skin against UVB-induced oxidative damage by using a three-dimensional tissue culture model of human epidermis. The endpoints of our experiments were cell viability, release of interleukin-1alpha and prostaglandin E(2) (well-known mediators of cutaneous inflammatory processes), accumulation in the epidermis of malondialdehyde/4-hydroxynonenal and protein carbonyl groups (derived by the oxidative damage respectively of lipids and proteins) and tissue redox balance (expressed by the levels of reduced glutathione, oxidized glutathione, glutathione peroxidase and glutathione reductase). Taken together, our findings demonstrate that the JW-E is an efficient botanical mixture able to prevent skin oxidative damage induced by UV-B exposure and may thus be a potential promising candidate as a skin photoprotective agent.

Allergic contact angioedema to benzoyl peroxide.

Benzoyl peroxide (BP) is commonly used for topical acne treatment and has long been known to be a weak allergen and a strong irritant. We report a case of a 26-year-old woman, who presented with an itchy erythematous reaction and strong oedema localized to the face. Two weeks before angioedema, the patient had started a new topical treatment with a gel containing 10% BP for acne. She was patch tested to European Standard Series, including BP 1% in white petrolatum, and to the 10% BP-containing gel previously used by herself, showing positivity on day 2 to BP 1% and to the 10% BP-containing gel. Factors that suggested an association between the severe angioedematous reaction and BP topical application include the strong reaction to BP in the patch-test, the temporal relationship, the complete resolution of symptoms after the drug was withdrawn and the absence of other identified explanations.

Levels and congener pattern of polychlorinated biphenyl and organochlorine pesticide residues in bluefin tuna (Thunnus thynnus) from the Straits of Messina (Sicily, Italy).

The aim of this study is to assess the accumulation of OCs and PCBs in Thunnus thynnus and to elucidate the suitability of this species as a bioindicator for monitoring contaminations of these compounds in the marine ecosystems of the Straits of Messina. This investigation was conducted on fat, liver and muscle samples of 14 T. thynnus collected during April 2004. Quantitative determination of OCs and PCBs in the various samples examined has been carried out using GC-ECD and GC-MS. The results obtained show the presence of low concentrations of p,p'-DDE and PCB congeners (138, 153 and 180) in all fat, liver and muscle samples caught in the Straits of Messina. Concentrations of p,p'-DDE and PCB congeners (138, 153 and 180) in all the samples examined were below MRLs (CE n. 97/41, 1999/65 and 1999/71).

Effect of polysaccharides from Opuntia ficus-indica (L.) cladodes on the healing of dermal wounds in the rat.

In traditional medicine extracts of polysaccharide-containing plants are widely employed for the treatment of skin and epithelium wounds and of mucous membrane irritation. The extracts of Opuntia ficus-indica cladodes are used in folk medicine for their antiulcer and wound-healing activities. The present study describes the wound-healing potential of two lyophilized polysaccharide extracts obtained from O. ficus-indica (L.) cladodes applied on large full-thickness wounds in the rat. When topically applied for 6 days, polysaccharides with a molecular weight (MW)>10(4)Da from O. ficus-indica cladodes induce a beneficial effect on cutaneous repair in this experimental model; in particular the topical application of O. ficus-indica extracts on skin lesions accelerates the reepithelization and remodelling phases, also by affecting cell-matrix interactions and by modulating laminin deposition. Furthermore, the wound-healing effect is more marked for polysaccharides with a MW ranging 10(4)-10(6)Da than for those with MW>10(6)Da, leading us to suppose that the fine structure of these polysaccharides and thus their particular hygroscopic, rheologic and viscoelastic properties may be essential for the wound-healing promoter activity observed.

The almond milk: a new approach to the management of cow-milk allergy/intolerance in infants.

AIM: Elimination of the offending food is imperative in the management of children with cow-milk allergy/intolerance (CMA/CMI). Herein we report the result of randomized clinical trial carried out to test the efficacy and safety of a new almond-based food (hereinafter named almond milk) in a group of infant with CMI/CMA. METHODS: A group of 52 infants aged 5 to 9 months and with documented CMI/CMA was enrolled and randomized to: almond milk (Group A, n=26); soy-based formula (Group B, n=13); protein hydrolysate-based formula (n=13). The main efficacy outcomes were the improvement in clinical symptoms and the decrease in serum levels of soluble CD30 (a potential marker for atopic disorders; sCD30). RESULTS: Elimination of the offending food and supplementation with a milk protein-free formula produced a considerable improvement of clinical manifestations within 5-12 days in all cases examined (at the onset of the study: 26.4+/-5.4 U/mL and 7.9+/-5.2 U/mL in IgE+ and IgE- infants respectively, after 6 months of supplementation: 16.6+/-4.8 U/mL and 7.1+/-4.5 U/mL in IgE+ and IgE- infants respectively). No difference in growth rate (increment of weight, length and head circumference) was found, during the entire study, between infants given the almond milk and babies given the soy-based formula or the protein hydrolysate-based formula. Supplementation with the soy-based and protein hydrolysate-based formulas caused the development, in some subjects, of a secondary sensitization (23% to soy-based and 15% protein hydrolysate-based formula), whereas supplementation with the almond milk did not. CONCLUSIONS: Though preliminary, the present findings seem to demonstrate that the almond milk may an efficacious substitute of cow milk in infants with CMA/CMI. One could speculate that some active principles contained in the almond milk could contribute to its beneficial effect observed in CMI/CMA-affected infants.