Facile biocatalytic access to 9-fluorenylmethyl polyglycosides: evaluation of antiviral activity on immunocompetent cells

The biological activities of a series of mono- and oligosaccharides (beta-xylosides and alpha-glucosides) of 9-fluorenylmethanol were investigated together with mono-beta-galactoside and beta-glucoside of this aglycone, produced by biocatalytic routes. By using marine glycoside hydrolases and inexpensive donors such as maltose or xylan, access to mono-alpha-glucoside or mono-beta-xyloside of 9-fluorenylmethanol was obtained. Additionally, interesting polyglycoside derivatives were isolated. Biological testing indicated that in vitro treatment with these carbohydrate derivatives may influence the balance of cytokines in the environment of human peripheral blood mononuclear cells (PBMC), restricting the harmful effect of herpes simplex type 2 replication. In fact, these carbohydrate derivatives tested in WISH cells did not show any significant antiviral activity.     

Food borne yeasts as DNA-bioprotective agents against model genotoxins

Yeasts isolated from Italian beverages and foods (wine and cheeses) were identified as Saccharomyces cerevisiae and Debaryomyces hansenii by sequencing the D1/D2 domain of the 26S rRNA gene and differentiated, at strain level, by microsatellite PCR fingerprinting and RAPD-PCR.All the strains showed antioxidant activity, as demonstrated by their ability to scavenge the free radical diphenyl-1-picrylhydrazyl (DPPH). Furthermore, tested strains revealed high in vitro inhibitory activity against two model genotoxins, 4-nitroquinoline-1-oxide (4-NQO) and N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), as showed by short-term methods with different target cells: SOS-Chromotest with Escherichia coli PQ37 and Comet assay with HT-29 enterocytes. High inhibitory activity towards 4-NQO was associated with cell viability, while heat-inactivated cells showed a reduced antigenotoxic capability. Surprisingly, high inhibition of MNNG genotoxicity was observed even with heat-treated cells. Moreover, the strains able to inhibit the genotoxins induced some changes in the spectroscopic properties of the original compound. This result perfectly agrees with the information obtained by the two bioassays.Interestingly, strains characterized for antioxidant and antigenotoxic properties, also presented acid-bile tolerance, indicating that food autochthonous yeasts could be expected to reach gut in viable form and thus prevent genotoxin DNA damage in situ.     

Antiproliferative and antioxidant effects on breast cancer cells of oleuropein and its semisynthetic peracetylated derivatives

Olive leaves extracts are a natural source of polyphenols, mainly oleuropein, widely considered to be potentially beneficial for health. This study focused on evaluation of the anti-tumoural activities of some oleuropein peracetylated derivatives, obtained with “green chemical” methodologies, against two human breast cancer cell lines. MCF-7 and T-47D cells were treated with oleuropein, peracetylated oleuropein, peracetylated aglycone and peracetylated hydroxytyrosol and the effects on growth and viability were investigated. Antioxidant effects were analysed after treatment with hydrogen peroxide. The peracetylated compounds exerted higher antiproliferative effects than oleuropein, by an arrest of cell cycle progression, associated with a strong antioxidant activity. Our results demonstrate that olive leaves, a by-product of olive manufacture, may provide a precious source of chemical derivatives, obtainable by peracetylation of oleuropein derivatives, which provide beneficial properties for human health.     

Molecular identification and osmotolerant profile of wine yeasts that ferment a high sugar grape must

The objective of this study was to examine the Saccharomyces and non-Saccharomyces yeast populations involved in a spontaneous fermentation of a traditional high sugar must (Vino cotto) produced in central Italy. Molecular identification of a total of 78 isolates was achieved by a combination of PCR-RFLP of the 5.8S ITS rRNA region and sequencing of the D1/D2 domain of the 26S rRNA gene. In addition, the isolates were differentiated by RAPD-PCR. Only a restricted number of osmotolerant yeast species, i.e. Candida apicola, Candida zemplinina and Zygosaccharomyces bailii, were found throughout all the fermentation process, while Saccharomyces cerevisiae prevailed after 15 days of fermentation. A physiological characterization of isolates was performed in relation to the resistance to osmotic stress and ethanol concentration. The osmotolerant features of C. apicola, C. zemplinina and Z. bailii were confirmed, while S. cerevisiae strains showed three patterns of growth in response to different glucose concentrations (2%, 20%, 40% and 60% w/v). The ability of some C. apicola and C. zemplinina strains to grow at 14% v/v ethanol is noteworthy. The finding that some yeast biotypes with higher multiple stress tolerance can persist in the entire winemaking process suggests possible future candidates as starter for Vino cotto production.     

Role of Etanercept and Infliximab on Nociceptive Changes Induced by the Experimental Model of Fibromyalgia

Background: Fibromyalgia is a clinical condition that affects 1% to 5% of the population. No proper therapy has been currently found. It has been described that inflammation plays a central role in the nerve sensitizations that characterize the pathology. Methods: This paper aimed to evaluate the efficacy of etanercept and infliximab in the management of pain sensitization. Fibromyalgia was induced by three injections once a day of reserpine at the dose of 1 mg/kg. Etanercept (3 mg/kg) and infliximab (10 mg/kg) were administered the day after the last reserpine injection and then 5 days after that. Behavioral analyses were conducted once a week, and molecular investigations were performed at the end of the experiment. Results: Our data confirmed the major effect of infliximab administration as compared to etanercept: infliximab administration strongly reduced pain sensitization in thermal hyperalgesia and mechanical allodynia. From the molecular point of view, infliximab reduced the activation of microglia and astrocytes and the expression of the purinergic P2X7 receptor ubiquitously expressed on glia and neurons. Downstream of the P2X7 receptor, infliximab also reduced p38-MAPK overexpression induced by the reserpine administration. Conclusion: Etanercept and infliximab treatment caused a significant reduction in pain. In particular, rats that received infliximab showed less pain sensitization. Moreover, infliximab reduced the activation of microglia and astrocytes, reducing the expression of the purinergic receptor P2X7 and p38-MAPK pathway.     

Fatty Acid Amide Hydrolase (FAAH) Inhibition Plays a Key Role in Counteracting Acute Lung Injury

Acute lung injury (ALI) is a group of lung illnesses characterized by severe inflammation, with no treatment. The fatty acid amide hydrolase (FAAH) enzyme is an integral membrane protein responsible for the hydrolysis of the main endocannabinoids, such as anandamide (AEA). In pre-clinical pain and inflammation models, increasing the endogenous levels of AEA and other bioactive fatty acid amides (FAAs) via genetic deletion or the pharmacological inhibition of FAAH produces many analgesic benefits in several different experimental models. To date, nobody has investigated the role of FAAH inhibition on an ALI mouse model. Mice were subjected to a carrageenan injection and treated orally 1 h after with the FAAH inhibitor URB878 dissolved in a vehicle consisting of 10% PEG-400, 10% Tween-80 and 80% saline at different doses: The inhibition of FAAH activity was able to counteract not only the CAR-induced histological alteration, but also the cascade of related inflammatory events. URB878 clears the way for further studies based on FAAH inhibition in acute lung pathologies.     

Lightly supervised alignment of subtitles on multi-genre broadcasts

This paper describes a system for performing alignment of subtitles to audio on multigenre broadcasts using a lightly supervised approach. Accurate alignment of subtitles plays a substantial role in the daily work of media companies and currently still requires large human effort. Here, a comprehensive approach to performing this task in an automated way using lightly supervised alignment is proposed. The paper explores the different alternatives to speech segmentation, lightly supervised speech recognition and alignment of text streams. The proposed system uses lightly supervised decoding to improve the alignment accuracy by performing language model adaptation using the target subtitles. The system thus built achieves the third best reported result in the alignment of broadcast subtitles in the Multi–Genre Broadcast (MGB) challenge, with an F1 score of 88.8%. This system is available for research and other non–commercial purposes through webASR, the University of Sheffield’s cloud–based speech technology web service. Taking as inputs an audio file and untimed subtitles, webASR can produce timed subtitles in multiple formats, including TTML, WebVTT and SRT.     

Programming by example: The human face of AI

It is argued that “human-centredness” will be an important characteristic of systems that learn tasks from human users, as the difficulties in inductive inference rule out learning without human assistance. The aim of “programming by example” is to create systems that learn how to perform tasks from their human users by being shown examples of what is to be done. Just as the user creates a learning environment for the system, so the system provides a teaching opportunity for the user, and emphasis is placed as much on facilitating successful teaching as on incorporating techniques of machine learning. If systems can “learn” repetitive tasks, their users will have the power to decide for themselves which parts of their jobs should be automated, and teach the system how to do them — reducing their dependence on intermediaries such as system designers and programmers. This paper presents principles for programming by example derived from experience in creating four prototype learners: for technical drawing, text editing, office tasks, and robot assembly. A teaching metaphor (a) enables the user to demonstrate a task by performing it manually, (b) helps to explain the learner's limited capabilities in terms of a persona, and (c) allows users to attribute intentionality. Tasks are represented procedurally, and augmented with constraints. Suitable mechanisms for attention focusing are necessary in order to control inductive search. Hidden features of a task should be made explicit so that the learner need not embark on the huge search entailed by hypothesizing missing steps.