No abstract

Spent coffee grounds (SCG) were extracted with an environmentally friendly procedure and analyzed to evaluate the recovery of relevant natural antioxidants for use as nutritional supplements, foods, or cosmetic additives. SCG were characterized in terms of their total phenolic content by the Folin-Ciocalteu procedure and antioxidant activity by the DPPH scavenging assay. Flavonoid content was also determined by a colorimetric assay. The total phenolic content was strongly correlated with the DPPH scavenging activity, suggesting that phenolic compounds are mainly responsible for the antioxidant activity of SCG. An UHPLC-PDA-TOF-MS system was used to separate, identify, and quantify phenolic and nonphenolic compounds in the SCG extracts. Important amounts of chlorogenic acids (CGA) and related compounds as well as caffeine (CAF) evidenced the high potential of SCG, a waste material that is widely available in the world, as a source of natural phenolic antioxidants.

RNAi screens have, to date, identified many genes required for mitotic divisions of Drosophila tissue culture cells. However, the inventory of such genes remains incomplete. We have combined the powers of bioinformatics and RNAi technology to detect novel mitotic genes. We found that Drosophila genes involved in mitosis tend to be transcriptionally co-expressed. We thus constructed a co-expression–based list of 1,000 genes that are highly enriched in mitotic functions, and we performed RNAi for each of these genes. By limiting the number of genes to be examined, we were able to perform a very detailed phenotypic analysis of RNAi cells. We examined dsRNA-treated cells for possible abnormalities in both chromosome structure and spindle organization. This analysis allowed the identification of 142 mitotic genes, which were subdivided into 18 phenoclusters. Seventy of these genes have not previously been associated with mitotic defects; 30 of them are required for spindle assembly and/or chromosome segregation, and 40 are required to prevent spontaneous chromosome breakage. We note that the latter type of genes has never been detected in previous RNAi screens in any system. Finally, we found that RNAi against genes encoding kinetochore components or highly conserved splicing factors results in identical defects in chromosome segregation, highlighting an unanticipated role of splicing factors in centromere function. These findings indicate that our co-expression–based method for the detection of mitotic functions works remarkably well. We can foresee that elaboration of co-expression lists using genes in the same phenocluster will provide many candidate genes for small-scale RNAi screens aimed at completing the inventory of mitotic proteins.

The end-to-end association of chromosomes through their telomeres has been observed in normal cells of certain organisms, as well as in senescent and tumor cells. The molecular mechanisms underlying this phenomenon are currently unknown. We show here that five independent mutant alleles in the Drosophila UbcDl gene cause frequent telomere-telomere attachments during both mitosis and male meiosis that are not seen in wild type. These telomeric associations involve all the telomeres of the D. melanogaster chromosome complement, albeit with different frequencies. The pattern of telomeric associations observed in UbcDl mutants suggests strongly that the interphase chromosomes of wild-type larval brain cells maintain a Rabl orientation within the nucleus, with the telomeres and centromeres segregated to opposite sides of the nucleus. The UbcDl gene encodes a class I ubiquitin-conjugating (E2) enzyme. This indicates that ubiquitin-mediated proteolysis is normally needed to ensure proper telomere behavior during Drosophila cell division. We therefore suggest that at least one of the targets of UbcDl ubiquitination is a telomere-associated polypeptide that may help maintain proper chromosomal orientation during interphase.

A method has been developed for the electrophoretic and quantitative analyses of human red cell delta-aminolevulinate dehydrase (ALADH). The enzyme is under the control of an autosomal gene, with two common codominant alleles. ALADH1 and ALADH2, with frequencies of 0.89 and 0.11, respectively, in the Italian population. Mean phenotypic enzyme activities are nearly identical: 52,. 49 and 55 mIU/g Hb for ALADH 1, 2-1 and 2 phenotypes respectively.

Many analytical methods for polyphenol determination in food and beverages can be found in the literature, but most of them need time-consuming sample pretreatment. Conversely, methods are missing for a rapid screening of non-pre-treated samples, with useful application in the agri-food industry, from process control to fraud. Selected ion recording mass mode after liquid chromatographic separation was used for the detection and quantification of free polyphenols in three craft beers, after just degassing, filtering, and diluting the beer samples with the mobile phase prior to the analysis. Fourteen polyphenols including hydroxybenzoic acids, hydroxycinnamic acids, and flavonols were chosen as standards. Nine of them were identified and quantified in at least one of the analyzed craft beers, despite the low content and the complexity of the samples. Depending on the analyte, 80− 7 μg/L as the limit of detection (LOD) and 240−30 μg/L as the limit of quantification (LOQ) were found. Satisfactory results for interday and intraday precisions, accuracy, recovery, and matrix effect were found. The one-factor analysis of variance (ANOVA) on the three different craft beers was statistically significant, with P < 0.05. The electrospray ionization mass fingerprinting of the same non-pre-treated craft beers was also investigated for wider characterization. Diagnostic ions were found and identified as deriving from oligosaccharides, organic acids, and amino acids.

Beyond the experimental methods used, the discussion of the present results might provide food for thought to the wide audience working on antioxidants.

Scavengers of hypochlorite (XOCl) could have beneficial effects in diseases in which this oxidant plays a pathogenic role. It has been reported that ferulic acid and chlorogenic acid, the quinic ester of caffeic acid, are good hypochlorite scavengers, but a systematic evaluation of the naturally occurring hydroxycinnamic acids (HCAs), which these substances belong to, has not been performed yet. Thus, in this work we studied, by two different in-vitro methods, the antioxidant activity of five HCAs: p-coumaric acid, ferulic acid, sinapinic acid, caffeic acid and chlorogenic acid. The methods applied in this study were based on the oxidation of human serum albumin (HSA) by XOCl, a new microplate method based on the measurement of chloramines and a previously described carbonyl assay. Firstly, lysine-derived chloramines, in the presence or absence of the HCAs, were detected using 5-thio-2-nitrobenzoic acid (TNB), measuring the absorbance at 415 nm by a microplate reader. To remove excess XOCl, Trolox, a known XOCl scavenger, was added before TNB. Secondly, lysine-derived carbonyls, in the presence or absence of the HCAs, were detected by using 2,4-dinitrophenylhydrazine. Hydroxycinnamic acids appeared active (caffeic >/= sinapinic > chlorogenic congruent with ferulic > p-coumaric acid) by both methods, suggesting possible pharmacological applications for these compounds, which are present at high concentrations in the plant kingdom.