Hydrolysis of 4-methylumbelliferyl butyrate: A convenient and sensitive fluorescent assay for lipase activity

A rapid, sensitive and convenient fluorescent assay was developed to screen for lipase activity. The non-fluorescent substrate, 4-methylumbelliferyl butyrate (4-MUB), solubilized in either liposomal dispersions or bile salt/lecithin mixed micelles, is hydrolyzed to butyric acid and the highly fluorescent compound, 4-methylumbelliferone (4-MU). Assays are run at 37 C for 10 min, terminated, and the changes in fluorescence quantitated with a Turner III fluorometer. Both lingual and pancreatic lipases exhibit activity against this artificial substrate. The assay has several advantages: nmoles 4 MU/ml/hr are measured allowing the detection of very low lipolytic activity; multiple samples may be simultaneously assayed, and only a brief incubation period is required.     

Mathematical determination of feeding patterns and its consequence on correlational studies

The cell cycle regulatory proteins, which include cyclin-dependent kinases (cdks), cdk inhibitors (CKIs), cyclins, and the pRB, and E2F families of proteins, constitute a network of interacting factors which govern exit from or passage through the mammalian cell cycle. While the proteins within these families have similar structural characteristics, each family member exhibits distinct expression patterns during embryogenesis and distinct biological activities. In order to begin to understand the tissue-specific roles of these interacting factors, we determined the expression pattern and activity of the pRB, E2F, cyclin, cdk, and CKI families of cell cycle regulatory proteins during retinoic acid-induced (neuronal pathway) and DMSO-induced (cardiac muscle pathway) differentiation of the pluripotent murine embryonal carcinoma cell line, P19. We demonstrate here that P19 terminal differentiation causes lineage-specific changes in the expression and activity of distinct members of the E2F, pRB, cyclin, and CKI families. Furthermore, dynamic changes in the activities of these cell cycle regulatory proteins occur through several overlapping mechanisms, culminating in repression of DNA-binding activity by all of the E2F family members as cells terminally differentiate.     

Structural transformations in surface layers of amorphous ribbons Ni78SixB22−x

The thermal stability of NI₇₈Si_xB_22–x (x = 6, 9, 12, 15) metallic glasses, produced by the single-roll melt spinning method, has been investigated by differential thermal analysis (DTA) and by measuring the temperature dependences of the exoelectron emission (EEE) and contact potential difference (CPD) relative to platinum. It has been found that both CPD and EEE show anomalies at temperatures corresponding to the structural transformations in the surface layer of investigated materials. The surface crystallization occurs at temperature of 150 K lower than the volume crystallization. The activation energy for the surface crystallization is also distinctly lower than that for crystallization in bulk.     

Glimmer: Multilevel MDS on the GPU

We present Glimmer, a new multilevel algorithm for multidimensional scaling designed to exploit modern graphics processing unit (GPU) hardware. We also present GPU-SF, a parallel, force-based subsystem used by Glimmer. Glimmer organizes input into a hierarchy of levels and recursively applies GPU-SF to combine and refine the levels. The multilevel nature of the algorithm makes local minima less likely while the GPU parallelism improves speed of computation. We propose a robust termination condition for GPU-SF based on a filtered approximation of the normalized stress function. We demonstrate the benefits of Glimmer in terms of speed, normalized stress, and visual quality against several previous algorithms for a range of synthetic and real benchmark datasets. We also show that the performance of Glimmer on GPUs is substantially faster than a CPU implementation of the same algorithm.     

Adaptive course generation through learning styles representation

This paper presents an approach to automatic course generation and student modeling. The method has been developed during the European funded projects Diogene and Intraserv, focused on the construction of an adaptive e-learning platform. The aim of the platform is the automatic generation and personalization of courses, taking into account pedagogical knowledge on the didactic domain as well as statistic information on both the student’s knowledge degree and learning preferences. Pedagogical information is described by means of an innovative methodology suitable for effective and efficient course generation and personalization. Moreover, statistic information can be collected and exploited by the system in order to better describe the student’s preferences and learning performances. Learning material is chosen by the system matching the student’s learning preferences with the learning material type, following a pedagogical approach suggested by Felder and Silverman. The paper discusses how automatic learning material personalization makes it possible to facilitate distance learning access to both able-bodied and disabled people. Results from the Diogene and Intraserv evaluation are reported and discussed.     

Scale-effects on mean and standard deviation of the mechanical properties of condensed matter: an energy-based unified approach

The size effects on the mean values of the mechanical properties of condensed matter and on the related variances are analysed by means of a unified approach based on the multiscale character of energy dissipation. In particular, the scaling law for fragmentation energy density is obtained taking into account the self-similarity of fragments. It is based on a generalization of the three classical comminution laws that has been performed to evaluate the energy dissipation, computing volume and surface area of the particles for one- two- and three-dimensional fragmented objects. The result is general and can be applied to different fractal energy dissipation mechanisms, e.g., plasticity. Based on this approach, the scaling laws for mean and standard deviation values of the main mechanical properties of materials can be derived, like Young's and shear elastic moduli, ultimate normal and shear stresses and strains, fracture energy and toughness.     

A Novel Human Neutralizing mAb Recognizes Delta,Gamma and Omicron Variants of SARS-CoV-2 and Can Be Used in Combination with Sotrovimab

The dramatic experience with SARS-CoV-2 has alerted the scientific community to be ready to face new epidemics/pandemics caused by new variants. Among the therapies against the pandemic SARS-CoV-2 virus, monoclonal Antibodies (mAbs) targeting the Spike glycoprotein have represented good drugs to interfere in the Spike/ Angiotensin Converting Enzyme-2 (ACE-2) interaction, preventing virus cell entry and subsequent infection, especially in patients with a defective immune system. We obtained, by an innovative phage display selection strategy, specific binders recognizing different epitopes of Spike. The novel human antibodies specifically bind to Spike-Receptor Binding Domain (RBD) in a nanomolar range and interfere in the interaction of Spike with the ACE-2 receptor. We report here that one of these mAbs, named D3, shows neutralizing activity for virus infection in cell cultures by different SARS-CoV-2 variants and retains the ability to recognize the Omicron-derived recombinant RBD differently from the antibodies Casirivimab or Imdevimab. Since anti-Spike mAbs, used individually, might be unable to block the virus cell entry especially in the case of resistant variants, we investigated the possibility to combine D3 with the antibody in clinical use Sotrovimab, and we found that they recognize distinct epitopes and show additive inhibitory effects on the interaction of Omicron-RBD with ACE-2 receptor. Thus, we propose to exploit these mAbs in combinatorial treatments to enhance their potential for both diagnostic and therapeutic applications in the current and future pandemic waves of coronavirus.     

Chronic Hyperkaliemia in Chronic Kidney Disease: An Old Concern with New Answers

Increasing potassium intake ameliorates blood pressure (BP) and cardiovascular (CV) prognoses in the general population; therefore the World Health Organization recommends a high-potassium diet (90–120 mEq/day). Hyperkalaemia is a rare condition in healthy individuals due to the ability of the kidneys to effectively excrete dietary potassium load in urine, while an increase in serum K⁺ is prevalent in patients with chronic kidney disease (CKD). Hyperkalaemia prevalence increases in more advanced CKD stages, and is associated with a poor prognosis. This scenario generates controversy on the correct nutritional approach to hyperkalaemia in CKD patients, considering the unproven link between potassium intake and serum K⁺ levels. Another concern is that drug-induced hyperkalaemia leads to the down-titration or withdrawal of renin-angiotensin system inhibitors (RASI) and mineralocorticoids receptors antagonists (MRA) in patients with CKD, depriving these patients of central therapeutic interventions aimed at delaying CKD progression and decreasing CV mortality. The new K⁺-binder drugs (Patiromer and Sodium-Zirconium Cyclosilicate) have proven to be adequate and safe therapeutic options to control serum K⁺ in CKD patients, enabling RASI and MRA therapy, and possibly, a more liberal intake of fruit and vegetables.     

Oxidative Stress and Chemoradiation-Induced Oral Mucositis: A Scoping Review of In Vitro,In Vivo and Clinical Studies

Chemoradiation-induced mucositis is a debilitating condition of the gastrointestinal tract eventuating from antineoplastic treatment. It is believed to occur primarily due to oxidative stress mechanisms, which generate Reactive Oxygen Species (ROS). The aim of this scoping review was to assess the role of oxidative stress in the development of Oral Mucositis (OM). Studies from the literature, published in MEDLINE and SCOPUS, that evaluated the oxidative stress pathways or antioxidant interventions for OM, were retrieved to elucidate the current understanding of their relationship. Studies failing inclusion criteria were excluded, and those suitable underwent data extraction, using a predefined data extraction table. Eighty-nine articles fulfilled criteria, and these were sub-stratified into models of study (in vitro, in vivo, or clinical) for evaluation. Thirty-five clinical studies evaluated antioxidant interventions on OM’s severity, duration, and pain, amongst other attributes. A number of clinical studies sought to elucidate the protective or therapeutic effects of compounds that had been pre-determined to have antioxidant properties, without directly assessing oxidative stress parameters (these were deemed “indirect evidence”). Forty-seven in vivo studies assessed the capacity of various compounds to prevent OM. Findings were mostly consistent, reporting reduced OM severity associated with a reduction in ROS, malondialdehyde (MDA), myeloperoxidase (MPO), but higher glutathione (GSH) and superoxide dismutase (SOD) activity or expression. Twenty-one in vitro studies assessed potential OM therapeutic interventions. The majority demonstrated successful a reduction in ROS, and in select studies, secondary molecules were assessed to identify the mechanism. In summary, this review highlighted numerous oxidative stress pathways involved in OM pathogenesis, which may inform the development of novel therapeutic targets.