Phenolic content and antioxidant activity of donkey milk kefir fortified with sulla honey and rosemary essential oil during refrigerated storage

The aim of this work was to evaluate the phenolic content and antioxidant activity of donkey kefirfortified with sulla honey and rosemary essential oil, during refrigerated storage. The type offlavouring strongly influenced the antioxidant activity of the kefir: rosemary essential oil kefirshowed the highest 2,20-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid values; sulla honey kefirshowed the highest ferric-reducing antioxidant power values but, at the same time, in both fortifiedkefirs, the thiol content decreased, probably because of the formation of polyphenol-protein com-plexes that would have influence the availability of bioactive components. The antioxidant activityincreased significantly during refrigerated storage, showing the highest values after 15 days. Sen-sory analysis highlighted the fact that the donkey kefir was well accepted by consumers. Addition ofsulla honey resulted in the highest acceptability, while addition of rosemary essential oil kefir wasless accepted by consumers. This knowledge provides a basis that could lead to the production offortified donkey kefir with specific nutraceutical characteristics

TF-ChIP Method for Tissue-Specific Gene Targets

Chromatin immunoprecipitation (ChIP) is an assay developed in order to define the dynamic nature of transcription processes. This method has been widely employed to identify methylated and acetylated DNA sequences in a variety of organs both in animals and humans. Nevertheless, this technique is significantly less employed to study transcriptional targets of specific nuclear signaling factors (TFs) and the data published so far have mainly used cell culture material and have been hardly reproduced in ex-vivo tissue. As nuclear signaling underlies important adaptive and maladaptive responses in chronic conditions such as cancer and neurodegeneration, there is a need for streamlining the upfront workflow of TF-ChIP for subsequent target sequencing. Based on the typical low concentration of the signaling transcriptional complex and the complexity/length of the ChIP Seq protocol, the field of cellular signaling has been confronted with a major roadblock in identifying clinically relevant targets of pathological and physiological signaling pathways. The present protocol offers a standardized procedure for detecting signaling targets in any whole tissue or specific dissected regions. The advantages of the protocol compared to the existing published methods are: (1) the small amount of starting material; appropriate for tissue subregions; (2) the optimization of DNA fragmentation from whole tissue; (3) suitability for sparsely populated tissues (i.e., brain); (4) the specificity of the TF- targeting readout; and (5) high DNA quality for sequencing or hybridization. The present protocol is highly detailed and can be reproduced using both fresh and fresh- frozen tissue. This is particularly relevant in the clinical setting, where specimen integrity is often the limiting step and where transcriptional target profiling is therapeutically relevant. The method is centered on Notch signaling but can be applied to a variety of nuclear signaling pathways as long as specific antibodies are available for pull down. Taken the superior yield/readout of this procedure, ChIP may finally provide relevant information about dynamic downstream gene changes in vivo for use in both basic research and clinical applications

Antioxidant activity of different cheese-honey combinations before and after in vitro gastrointestinal digestion

This study aimed to evaluate the effect of different cheese-honey combinations on antioxidant activity before and after in vitro gastrointestinal digestion, using the 2,2′-azino-bis-3-ethylbenzothiazoline- 6-sulfonic acid (ABTS), and ferric-reducing antioxidant power (FRAP) assays. Honey addition significantly enhanced the antioxidant activity of cheese samples (P < 0.05). The antioxidant activity of in vitro digested samples was significantly lower than the undigested ones. However, digested cheese-honey combination showed a percentage decrease of antioxidant activity lower than plain cheese. The cheese and honey types affected the percentage decrease of antioxidant activity in the digested samples, this is mainly due to the bioaccessibility of the honey polyphenols after digestion. Sensory analysis showed that all cheese-honey combinations were well accepted by the consumers, the health information on higher antioxidant capacity of dark honeys has not influenced the degree of acceptability of consumers, who preferred the cheese - clear honey combination

Antioxidant Properties, Polyphenol Content and Colorimetric Characteristics of Different Floral Origin Honeys from Different Areas of Southern Italy.

Ninety honey samples of five different floral origins (chestnut, eucalyptus, citrus, multifloral and sulla) from nine areas of southern Italy, were screened to evaluate the polyphenol content by spectrphotometric methods, the antioxidant activity by ABTS (2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)) diamonium salt, FRAP (ferric reducing antioxidant power) and DPPH (1,1-diphenyl-2-picrylhydrazyl) assays, and the colorimetric characteristics by spectrophotometric and CIE (Commission Internationale de l'Eclairage) L*a*b* methods. Moreover, the correlations between the analysed parameters were studied. Overall, the results showed that all the honey samples presented high total phenolic and flavonoid contents (12.23 mg gallic acid equivalent/100g honey and 8.16 mg quercetin equivalent/100g honey, respectively), and a high antioxidant activity (59.17%, 66.50% and 349.11 μM Fe (II) for ABTS, DPPH and FRAP assays, respectively), but those results differ widely according to the honey types, suggesting that honey composition has been influenced by its floral and geographical origin. In particular, the darkest honeys, such as chestnut honey (L* = 59.94; 1.26 AU), showed the highest polyphenol content and the highest antioxidant activity. Honeys from the areas with major anthropogenic activities and high population density presented the highest antioxidant activity. Correlations between the analysed parameters were statistically significant (P < 0.05), suggesting that the polyphenol content is correlated with the antioxidant activity and colour of honey

Metals in Honeys from Different Areas of Southern Italy

The aim of the study was to quantify the cadmium (Cd), lead (Pb), chromium (Cr) and arsenic (As) contents in ninety honey samples from nine areas of southern Italy. Results showed that As content was below the detection limit, while Cd, Pb, and Cr contents were below the recommended maximum acceptable levels. Mean Cd, Pb, and Cr contents were 0.013, 0.289 and 0.707 mg kg-1, respectively. The metal contents in honey varied greatly depending on considered area. Correlations between the metals were statistically significant (p\0.05), suggesting that polluting sources involve the simultaneous presence of metals in honey

A comparative study on phenolic profile, vitamin C content and antioxidant activity of Italian honeys of different botanical origin

The aim of our study was to identify and quantify the phenolic acids, flavonoids and vitamin C and to evaluate the antioxidant activity in ninety Italian honeys of different botanical origins (chestnut, sulla, eucalyptus, citrus and multifloral). The results showed that total phenolic and flavonoid contents varied from 11.08 to 14.26 mg GAE per 100 g honey and from 5.82 to 12.52 mg QE per 100 g honey, respectively. HPLC–UV analysis showed a similar but quantitatively different phenolic profile of the studied honeys. Vitamin C is present in all samples. Multifloral honey showed the highest amount of the detected total phenolic compounds and the highest vitamin C content. The DPPH value varied from 55.06 to 75.37%. Among the unifloral honeys, chestnut honey presented the highest levels of phenolic acids, flavonoids and vitamin C, which are closely associated with its high antioxidant activity. The results show that honey contains high amount of biologically active compounds, which play an important role in defining the nutraceutical quality of the product, and that the distribution of these compounds is influenced by the botanical origin

Effect of species on the distribution and oxidative stability of milk added of lead and cadmium

The aim of this study was to evaluate the effect of species on added lead (Pb) and cadmium (Cd) content in cow, buffalo, and goat milk and their distribution in fat, casein, and whey fractions. In addition, the oxidative stability of the milk was evaluated. Most of the Pb and Cd were recovered in the skimmed milk (96.74 and 94.21%, respectively). The distribution of Cd and Pb in casein and whey fractions, obtained by enzymatic coagulation, highlighted that they were mainly associated with casein (on average 94.77 and 90.54% of Pb and Cd, respectively). The species significantly affected the distribution of Cd and Pb in the casein and fat fractions (p &lt; 0.01). In particular, Cd and Pb levels in fat fraction were the highest in the buffalo milk, whereas casein fraction was the highest in bovine milk. Furthermore, the presence of metals negatively influenced the oxidative stability of the milk and the species influenced its response. The results showed that in the presence of Cd and Pb, bovine milk increased the content of Malondialdehyde (MDA), advanced oxidation protein products (AOPP) and dithyrosines compared to other species. In addition, there was a significant decrease in the thiol content, highlighting a reduction in the antioxidant capacity of the contaminated milk

Minerals content in Basilicata region (southern Italy) honeys from areas with different anthropic impact

SummaryThe aim of this study was to examine the minerals content (toxic elements, macro‐elements and trace elements) of Basilicata region (southern Italy) honeys and compare the mineral profile of honeys classified as rural, industrial and urban according to anthropic characteristics of geographical origin. Overall, Ca, Na, Mg, Fe, Al, Zn and Mn were the most abundant elements detected in honeys, with average contents exceeding 1 ppm, whereas heavy metals content was lower than the maximum limit established for honey. Statistical analysis revealed significant differences (P < 0.05) among honeys as function of anthropic characteristics of geographical origin, with the exception for Se, Co and Ag content. Industrial honeys were characterised by the highest Zn, Cr, Sn, Cd and Pb content, urban honeys showed the highest As, Fe, Ni, Mn, Na, Mg and Ca content, whereas rural honeys showed the highest Cu, Al and Ba content (P < 0.05). The findings of this study highlighted that honeys mineral profile is closely related to different content of elements in environment, which is affected by anthropogenic activities

Fatty acids composition, cholesterol and vitamin E contents of Longissimus dorsi and Semitendinosus muscles of Suino Nero Lucano pigs slaughtered at two different weights

The nutritional quality of the lipid fraction of two muscles (Longissimus dorsi and Semitendinous) from Italian autochthonous genotype Suino Nero Lucano pigs slaughtered at two different weights was evaluated. Meat of Suino Nero Lucano pig showed a relatively low content of cholesterol and a higher proportion of unsaturated (UFA) than saturated fatty acids (SFA). Total cholesterol content was influenced by muscle, being higher in Longissimus dorsi (LD) than in Semitendinous (ST) muscle. No significant effects related to slaughter weight or muscle were found regarding vitamin Econtent. Slaughter weight strongly influenced n-3 and n-6 polyunsaturated fatty acids (PUFA) contents that decreased with increasing weight, and consequently, PUFA/SFA ratio. Muscle markedly influenced the contents of SFA, monounsaturated fatty acids (MUFA), and PUFA, and the dietetic properties of the meat. ST muscle, compared with the LD muscle, showed higher PUFA/SFA and PUFA n-6/PUFA n-3 ratios, and lower atherogenic and thrombogenic indices

TF-ChIP Method for Tissue-Specific Gene Targets

Chromatin immunoprecipitation (ChIP) is an assay developed in order to define the dynamic nature of transcription processes. This method has been widely employed to identify methylated and acetylated DNA sequences in a variety of organs both in animals and humans. Nevertheless, this technique is significantly less employed to study transcriptional targets of specific nuclear signaling factors (TFs) and the data published so far have mainly used cell culture material and have been hardly reproduced in ex-vivo tissue. As nuclear signaling underlies important adaptive and maladaptive responses in chronic conditions such as cancer and neurodegeneration, there is a need for streamlining the upfront workflow of TF-ChIP for subsequent target sequencing. Based on the typical low concentration of the signaling transcriptional complex and the complexity/length of the ChIP Seq protocol, the field of cellular signaling has been confronted with a major roadblock in identifying clinically relevant targets of pathological and physiological signaling pathways. The present protocol offers a standardized procedure for detecting signaling targets in any whole tissue or specific dissected regions. The advantages of the protocol compared to the existing published methods are: (1) the small amount of starting material; appropriate for tissue subregions; (2) the optimization of DNA fragmentation from whole tissue; (3) suitability for sparsely populated tissues (i.e., brain); (4) the specificity of the TF-targeting readout; and (5) high DNA quality for sequencing or hybridization. The present protocol is highly detailed and can be reproduced using both fresh and fresh-frozen tissue. This is particularly relevant in the clinical setting, where specimen integrity is often the limiting step and where transcriptional target profiling is therapeutically relevant. The method is centered on Notch signaling but can be applied to a variety of nuclear signaling pathways as long as specific antibodies are available for pull down. Taken the superior yield/readout of this procedure, ChIP may finally provide relevant information about dynamic downstream gene changes in vivo for use in both basic research and clinical applications