Combining 1,3-Ditriazolylbenzene and Quinoline to Discover a New G-Quadruplex-Interactive Small Molecule Active against Cancer Stem-Like Cells

Quadruplex nucleic acids are promising targets for cancer therapy. In this study we used a fragment-based approach to create new flexible G-quadruplex (G4) DNA-interactive small molecules with good calculated oral drug-like properties, based on quinoline and triazole heterocycles. G4 melting temperature and polymerase chain reaction (PCR)-stop assays showed that two of these compounds are selective G4 ligands, as they were able to induce and stabilize G4s in a dose- and DNA sequence-dependent manner. Molecular docking studies have suggested plausible quadruplex binding to both the G-quartet and groove, with the quinoline module playing the major role. Compounds were screened for cytotoxicity against four cancer cell lines, where 4,4′-(4,4′-(1,3-phenylene)bis(1H-1,2,3-triazole-4,1-diyl))bis(1-methylquinolin-1-ium) ( 1 d ) showed the greater activity. Importantly, dose–response curves show that 1 d is cytotoxic in the human colon cancer HT-29 cell line enriched in cancer stem-like cells, a subpopulation of cells implicated in chemoresistance. Overall, this study identified a new small molecule as a promising lead for the development of drugs targeting G4 in cancer stem cells.     

Biogenic amines assessment during different stages of the canning process of skipjack tuna (Katsuwonus pelamis)

The present research focused on the biogenic amines (BAs) formation in skipjack tuna (Katsuwonus pelamis) throughout the whole canning process. In agreement with its wide employment on this species, on‐board brine immersion freezing (BIF) was tested as post‐mortem processing. The study included fish samples corresponding to different stages of the canning process such as frozen, thawed, cooked and canned; after cooking, two kinds of tuna muscles were considered, that is, whole fillets (main product) and grated muscle (off‐product arising from small pieces). For the BAs (tryptamine, putrescine, cadaverine, histamine, spermidine and spermine) assessment, an HPLC‐DAD method was developed and validated in skipjack tuna samples, in agreement with different parameters such as suitability, linearity, limits of detection and quantification, precision, accuracy and robustness. Tuna submitted on‐board to BIF procedure provided low levels of spermine and spermidine (up to 27.6 mg kg^?1), while contents on the remaining BAs maintained below the limit of detection. Throughout the different stages of the canning process, skipjack tuna showed a low formation of most BAs; interestingly, histamine content was found below 10.6 mg kg^?1 level. The highest values were obtained for spermidine, these related to cooked grated tuna (from 22.6 to 66.7 mg kg^?1) and canned grated tuna (from 70.6 to 104.4 mg kg^?1). Values for pH assessment in all kinds of tuna samples corroborated the results obtained for BAs determination. BIF procedure proved to be an amenable post‐mortem processing to guarantee the quality of canned skipjack tuna.     

Development of a long‐life vacuum‐packaged ready‐to‐eat meat product based on a traditional Portuguese seasoned meat

Carne do alguidar is a Portuguese traditional pork fried meat, usually manufactured for self‐consumption purposes. This study developed a ready‐to‐eat (RTE) meat product, to meet today's consumers’ convenience, manufactured at the industrial scale evaluating its quality and shelf life, assessing the effect of vacuum packaging and the use of an antioxidant (50 ppm BHT) to enhance oxidative stability. Physicochemical and microbiological parameters were assessed, and a sensory analysis was performed. Interestingly, no significant differences were recorded between control (non‐BHT) and antioxidant (BHT) samples. Microbiological counts remained at low levels throughout the storage period, ensuring the product's required microbiological quality. At later storage stages, higher values of thiobarbituric acid‐reactive substances arose and off flavours and aromas were perceived. Still, overall appreciation was not affected until 12 months of storage and a significant depreciation was perceived only after 15 months. Fibrousness and rising of off flavours were negatively correlated with overall appreciation.     

Carotenoids are Effective Inhibitors of in vitro Hemolysis of Human Erythrocytes,as Determined by a Practical and Optimized Cellular Antioxidant Assay

β‐Carotene, zeaxanthin, lutein, β‐cryptoxanthin, and lycopene are liposoluble pigments widely distributed in vegetables and fruits and, after ingestion, these compounds are usually detected in human blood plasma. In this study, we evaluated their potential to inhibit hemolysis of human erythrocytes, as mediated by the toxicity of peroxyl radicals (ROO?). Thus, 2,2′‐azobis (2‐methylpropionamidine) dihydrochloride (AAPH) was used as ROO? generator and the hemolysis assay was carried out in experimental conditions optimized by response surface methodology, and successfully adapted to microplate assay. The optimized conditions were verified at 30 × 10⁶ cells/mL, 17 mM of AAPH for 3 h, at which 48 ± 5% of hemolysis was achieved in freshly isolated erythrocytes. Among the tested carotenoids, lycopene (IC₅₀ = 0.24 ± 0.05 μM) was the most efficient to prevent the hemolysis, followed by β‐carotene (0.32 ± 0.02 μM), lutein (0.38 ± 0.02 μM), and zeaxanthin (0.43 ± 0.02 μM). These carotenoids were at least 5 times more effective than quercetin, trolox, and ascorbic acid (positive controls). β‐Cryptoxanthin did not present any erythroprotective effect, but rather induced a hemolytic effect at the highest tested concentration (3 μM). These results suggest that selected carotenoids may have potential to act as important erythroprotective agents by preventing ROO?‐induced toxicity in human erythrocytes.     

Validation of Calibration Methods at NMIs

The National Measurement Institutes (NMIs) topped off its country national measurement system transfer by calibration the traceability to the SI units. In order to fulfill the ISO/IEC 17025 standard, namely point 5.4.5, these laboratories should validate their own laboratory-designed or developed methods for the dissemination of the units. A procedure for validation of calibration methods is proposed and an example of validation of calibration results is drafted by the application of experimental design.     

The EU data protection reform and the challenges of big data: remaining uncertainties and ways forward

As the first broad reform of the EU data protection legislation is being achieved, and notwithstanding EU institutions’ confident discourse, scepticism remains about the reform’s ability to safeguard the fundamental right to data protection in the face of evolving data processing techniques underlying so-called big data. Yet, one might wonder whether the cause for this difficulty should be ascribed mainly to technological progresses that the law finds it hard to deal with or rather to the policy choices embedded in the legal reform itself. Indeed, a new data protection enforcement model is being adopted, which relies heavily on risk assessment and management by the data controllers themselves. Likewise, data protection authorities see their supervisory role significantly weakened. These developments and the underlying rationality are discussed. Given the limitations of the risk-based approach as currently devised, we suggest that it be reappraised in consideration of risk regulatory experiences in other domains.     

Antibiotic resistance in coagulase negative staphylococci isolated from wastewater and drinking water

This study reports the antibiotic resistance patterns of coagulase negative staphylococci (CNS) isolated from a drinking water treatment plant (WTP), a drinking water distribution network, responsible for supplying water to the consumers (WDN), and a wastewater treatment plant (WWTP), responsible for receiving and treating domestic residual effluents. Genotyping and the 16S rRNA gene sequence analysis demonstrated a higher diversity of species both in the WTP (6 species/19 isolates) and WWTP (12 species/47 isolates) than in the WDN (6 species/172 isolates). Staphylococcus pasteuri and Staphylococcus epidermidis prevailed in the WTP and WDN and Staphylococcus saprophyticus in the WWTP. Staphylococci with reduced susceptibility (resistance or intermediary phenotype) to beta-lactams, tetracycline, clindamycin and erythromycin were observed in all types of water and belonged to the three major species groups. The highest resistance rate was found against erythromycin, presumably due to the presence of the efflux pump encoded by the determinant msrA, detected in the majority of the resistant isolates. This study demonstrates that antibiotic resistant CNS may colonize different types of water, namely drinking water fulfilling all the quality standards.     

Novel features for time series analysis: a complex networks approach

Data Mining and Knowledge Discovery - Being able to capture the characteristics of a time series with a feature vector is a very important task with a multitude of applications, such as...