Functional role of high-affinity anandamide transport, as revealed by selective inhibition

Anandamide, an endogenous ligand for central cannabinoid receptors, is released from neurons on depolarization and rapidly inactivated. Anandamide inactivation is not completely understood, but it may occur by transport into cells or by enzymatic hydrolysis. The compound N-(4-hydroxyphenyl)arachidonylamide (AM404) was shown to inhibit high-affinity anandamide accumulation in rat neurons and astrocytes in vitro, an indication that this accumulation resulted from carrier-mediated transport. Although AM404 did not activate cannabinoid receptors or inhibit anandamide hydrolysis, it enhanced receptor-mediated anandamide responses in vitro and in vivo. The data indicate that carrier-mediated transport may be essential for termination of the biological effects of anandamide, and may represent a potential drug target.     

Neocytoapheresis in the treatment of polycythemia vera

In absence of thrombocytosis, periodic bloodlettings represent the elective therapy of polycythemia vera. In the present study we tested if neocytoapheresis, a method able to remove large quantities of younger, and then bigger red cells could represent an alternative therapeutic choice in these patients. We found that the employment of neocytoapheresis produced a remarkable delay in the time of procedures with a mean interval of 100 +/- 55 days. The mean value of hematocrit before neocytopheresis is resulted statistically different in comparison with the hematocrit after the procedure (p = 0.0001). The reticulocyte count is resulted higher in apheresis product in comparison with the blood control measured before procedure (p = 0.0001). In the same way, the mean corpuscular volume in the collection bags was higher than the volume measured before neocytoapheresis (p = 0.0095). We did not find any variation about the mean values of white blood cells and platelets before and after neocytoapheresis in the patients examined. These preliminary data seem to underline a better withdrawal of big size cells by this technique suggesting the efficacy of neocytoapheresis in the treatment of polycythemia vera.     

Microalbuminuria in IDDM is associated with increased expression of monocyte procoagulant activity

Insulin-like growth factor II (IGF-II) plays a key role in mammalian growth, influencing foetal cell division and differentiation and possibly metabolic regulation. The mature 67 amino acid peptide shares sequence homology with both insulin and IGF-I. The liver is the main endocrine source of IGFs, but autocrine/paracrine activity is found in most tissues. The type 1 receptor mediates most of the biological effects of IGF-I and IGF-II; the type 2 receptor is involved with IGF-II degradation. Binding proteins may both localise IGFs to the receptors and regulate their activities. The IGF2 gene is maternally imprinted in mouse and human. Relaxation of IGF2 imprinting occurs in the Beckwith-Wiedemann syndrome of somatic overgrowth, sporadic Wilms' tumour and a number of other cancers. In the general adult population, the IGF2-INS gene cluster may also influence body weight, in which case IGF-II function could become a target for therapeutic intervention in obesity.     

The impact of risk and protective factors on online teaching experience in high school Italian teachers during the COVID-19 pandemic

The lockdown due to COVID-19 in Italy resulted in the sudden closure of schools, with a shift from traditional teaching to the online one. Through an online questionnaire, this survey explores teachers' experience of online teaching, the level of risk factors (e.g., stress) and protective factors (e.g., locus of control) and their impact on satisfaction levels during the social distancing. One hundred seven high school teachers from Lombardy, an Italian region very affected by the COVID-19 outbreak, participated. Results show that depression and stress are the main predictors of satisfaction levels for online teaching. In addition, coping, locus of control and self-efficacy emerge as important protective factors. Finally, although there is great satisfaction with the online teaching experience, critical elements emerged. This study is relevant because it describes the critical elements of the online teaching experience, and identifies some protective factors and the main risk factors in teachers operating in an area strongly marked by social restrictions imposed by the pandemic. High school teachers emerge as a sub-group of the general population with specific psychological reactions. Considering the results, it is possible to suggest providing high-quality educational support and crisis-psychological oriented services to teachers, and help to maintain the psychological well-being.     

The GAUGAA Motif Is Responsible for the Binding between circSMARCA5 and SRSF1 and Related Downstream Effects on Glioblastoma Multiforme Cell Migration and Angiogenic Potential

Circular RNAs (circRNAs) are a large class of RNAs with regulatory functions within cells. We recently showed that circSMARCA5 is a tumor suppressor in glioblastoma multiforme (GBM) and acts as a decoy for Serine and Arginine Rich Splicing Factor 1 (SRSF1) through six predicted binding sites (BSs). Here we characterized RNA motifs functionally involved in the interaction between circSMARCA5 and SRSF1. Three different circSMARCA5 molecules (Mut1, Mut2, Mut3), each mutated in two predicted SRSF1 BSs at once, were obtained through PCR-based replacement of wild-type (WT) BS sequences and cloned in three independent pcDNA3 vectors. Mut1 significantly decreased its capability to interact with SRSF1 as compared to WT, based on the RNA immunoprecipitation assay. In silico analysis through the “Find Individual Motif Occurrences” (FIMO) algorithm showed GAUGAA as an experimentally validated SRSF1 binding motif significantly overrepresented within both predicted SRSF1 BSs mutated in Mut1 (q-value = 0.0011). U87MG and CAS-1, transfected with Mut1, significantly increased their migration with respect to controls transfected with WT, as revealed by the cell exclusion zone assay. Immortalized human brain microvascular endothelial cells (IM-HBMEC) exposed to conditioned medium (CM) harvested from U87MG and CAS-1 transfected with Mut1 significantly sprouted more than those treated with CM harvested from U87MG and CAS-1 transfected with WT, as shown by the tube formation assay. qRT-PCR showed that the intracellular pro- to anti-angiogenic Vascular Endothelial Growth Factor A (VEGFA) mRNA isoform ratio and the amount of total VEGFA mRNA secreted in CM significantly increased in Mut1-transfected CAS-1 as compared to controls transfected with WT. Our data suggest that GAUGAA is the RNA motif responsible for the interaction between circSMARCA5 and SRSF1 as well as for the circSMARCA5-mediated control of GBM cell migration and angiogenic potential.     

Resolution proof transformation for compression and interpolation

Verification methods based on SAT, SMT, and theorem proving often rely on proofs of unsatisfiability as a powerful tool to extract information in order to reduce the overall effort. For example a proof may be traversed to identify a minimal reason that led to unsatisfiability, for computing abstractions, or for deriving Craig interpolants. In this paper we focus on two important aspects that concern efficient handling of proofs of unsatisfiability: compression and manipulation. First of all, since the proof size can be very large in general (exponential in the size of the input problem), it is indeed beneficial to adopt techniques to compress it for further processing. Secondly, proofs can be manipulated as a flexible preprocessing step in preparation for interpolant computation. Both these techniques are implemented in a framework that makes use of local rewriting rules to transform the proofs. We show that a careful use of the rules, combined with existing algorithms, can result in an effective simplification of the original proofs. We have evaluated several heuristics on a wide range of unsatisfiable problems deriving from SAT and SMT test cases.     

Structures and allelopathic effects ofNuphar alkaloids: Nupharolutine and 6,6′-dihydroxythiobinupharidine

Two of the majorNuphar alkaloids, nupharolutine and 6,6-dihydroxythiobinupharidine, were isolated from the aquatic perennial herbNuphar lutea (L.) Sibth. & Sm., yellow water lily. In a lettuce seedling bioassay of the two pure compounds, the former was inactive and the latter was highly inhibitory of radicle elongation at concentrations greater than 2 ppm. Structures and stereochemistry of the two compounds were confirmed by DEPT,¹H-¹H COSY,¹H-¹³C COSY, and¹H-¹H NOESY experiments.     

Preparation and characterization of comb-shaped polyesters from 2,2-dioctadecyl-1,3-propanediol and phthalic acids

Summary New types of comb-shaped polyesters have been prepared by transesterification of 2,2-dioctadecyl-1,3-propanediol and the three isomeric diphenyl phthalates. Intrinsic viscosity and SEC analyses allowed determination of DPs (60–89) and molar masses; results compare well with absolute average weight molar masses (56,000–118,000) determined by low-angle laser light scattering. High-resolution ¹³C NMR reveals the characteristic aromatic and glycol polyester patterns; it resolves many side-chain methylene carbons, but shows no evidence of end-groups. All the polyesters are crystalline as shown by X-ray diffractometry and DSC, the crystallinity being exclusively due to the octadecyl side-group crystallization in the hexagonal form. The results indicate that the crystallinity decreases substantially when going from the ortho- to the terephthalate polyester.