Growth factor-induced Ca2+ responses are differentially modulated by nitric oxide via activation of a cyclic GMP-dependent pathway

Nitric oxide (NO) plays a modulatory role on cell growth and differentiation, biological processes that occur under the control of various signal transduction mechanisms, including those triggered by activation of membrane receptors for polypeptide growth factors. The increases in intracellular Ca2+ concentration elicited by the activation of these receptors are sustained by release of the cation from intracellular stores and by stimulation of this influx from the extracellular medium. Using NIH 3T3 cells overexpressing the human epidermal growth factor receptor, we investigated both of these processes stimulated by the administration of epidermal and platelet-derived growth factors as the receptor agonists. Pharmacological and functional analyses carried out on Fura-2-loaded cells showed that Ca2+ influx elicited by both growth factors is the summation of two distinct pathways, with the major pathway dependent on and the minor pathway independent of store depletion. Exposure of the cells to either No donors or NO synthase inhibitors induced increase and inhibition, respectively, of the two components of Ca2+ influx. When Ca2+ release was investigated, the above drugs were also active but in the opposite direction. The effects of NO were mimicked by the cGMP analogue 8-Br-cGMP and abolished by two cGMP-dependent protein kinase I inhibitors, whereas the cAMP analogue 8-Br-cAMP and two protein kinase A inhibitors had no appreciable effects. In addition, growth factors induced an increase in cGMP formation, an effect that was prevented by NO synthase inhibitors. In conclusion, NO appears to exert a feedback modulatory control on CA2+ responses to growth factor administration. Such a control might contribute to the inhibitory effect of NO on growth previously reported with various cell types.     

"No allogeneic blood transfusion" protocol for the surgical correction of craniosynostoses. II. Clinical application

The authors describe the results obtained in 13 consecutive cases of craniosynostosis operated on according to a protocol devised at avoiding allogeneic blood transfusion. The protocol is based on pre- and postoperative treatment with erythropoietin, preoperative autologous blood donation, preoperative normovolemic hemodilution and intraoperative blood salvage. Nine subjects were affected by simple forms of craniosynostosis, whereas the remaining 4 presented with oxycephaly or craniofacial syndromes. Five of the 13 children were under 7 months and a further 3, under 10 months of age at the time of the surgical operation. Seven children weighed less than 10 kg. Allogeneic blood transfusion was avoided in 11 of the 13 children considered. Two failures - defined as the necessity to reinfuse the patient with an allogeneic blood transfusion - were recorded, 1 of them resulting from an unexpected hemorrhage during surgery. The results obtained indicate that this protocol designed to avoid allogeneic blood transfusion can be safely applied in the great majority of children with craniosynostosis, even when the surgical correction is carried out early in life.     

Electrocortical desynchronization after microinfusion of kainic acid into the locus coeruleus in rats

CD-1, a genetically-engineered CHO cell line, was cultivated with a Biosilon microcarrier culture system. We successfully cultivated CD-1 cells to a very high density (over 1 x 10(7) cells/ml). Prourokinase was stably secreted at about 180 IU/10(6) cells/24 h. Experiments showed that CD-1 cells growing on Biosilon microcarriers were able to spontaneously release from the microcarriers, then reattach and proliferate on fresh microcarriers. This makes it very easy to scale up production. The microcarriers could be reused several times without affecting adhesion, proliferation and prourokinase secretion. With CM-PECC membrane radial flow chromatography and MPG chromatography, the prourokinase in conditioned medium could be purified to a specific activity of 1 x 10(5) IU/mg of protein. The purification factor was about 600 fold, and approximately 90% of the biological activity was recovered.     

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.     

Promoting the integration of technology in teaching: An analysis of the factors that increase the intention to use technologies among Italian teachers

Background: The rapid spread of COVID-19 forced many countries to adopt severe containment measures, transferring all didactic activities into virtual environments. However, the integration of technology in teaching may present difficulties, especially in some countries, such as Italy. Objectives: The present study analyzed how the two main factors of the Technology Acceptance Model (TAM), namely perceived usefulness and perceived ease of use, combined with online teaching self-efficacy, were associated with the intention to use technology to teach. We posited a moderated moderation model in which perceived usefulness represented the main predictor, with self-efficacy and perceived ease of use as moderators of intention to use technology to teach. Method: The model was tested through multiple regressions, using the PROCESS macro on SPSS 26 with a sample of 178 upper secondary school teachers in Italy. Results and conclusions: Regressions showed that each variable significantly predicted the intention to use technology. In addition, a moderation effect of self-efficacy on the perceived usefulness of using technology was found for medium and high-level of perceived ease of use of technology. Implications: The present study provides targeted implications for distance education policy and practice to promote its adoption (or the blended modality) in Italian upper schools.     

Neuroprotective Effects of Testosterone in the Hypothalamus of an Animal Model of Metabolic Syndrome

Metabolic syndrome (MetS) is known to be associated to inflammation and alteration in the hypothalamus, a brain region implicated in the control of several physiological functions, including energy homeostasis and reproduction. Previous studies demonstrated the beneficial effects of testosterone treatment (TTh) in counteracting some MetS symptoms in both animal models and clinical studies. This study investigated the effect of TTh (30 mg/kg/week for 12 weeks) on the hypothalamus in a high-fat diet (HFD)-induced animal model of MetS, utilizing quantitative RT-PCR and immunohistochemical analyses. The animal model recapitulates the human MetS features, including low testosterone/gonadotropin plasma levels. TTh significantly improved MetS-induced hypertension, visceral adipose tissue accumulation, and glucose homeostasis derangements. Within hypothalamus, TTh significantly counteracted HFD-induced inflammation, as detected in terms of expression of inflammatory markers and microglial activation. Moreover, TTh remarkably reverted the HFD-associated alterations in the expression of important regulators of energy status and reproduction, such as the melanocortin and the GnRH-controlling network. Our results suggest that TTh may exert neuroprotective effects on the HFD-related hypothalamic alterations, with positive outcomes on the circuits implicated in the control of energy metabolism and reproductive tasks, thus supporting a possible role of TTh in the clinical management of MetS.     

The AFLATOX® Project: Approaching the Development of New Generation,Natural-Based Compounds for the Containment of the Mycotoxigenic Phytopathogen Aspergillus flavus and Aflatoxin Contamination

The control of the fungal contamination on crops is considered a priority by the sanitary authorities of an increasing number of countries, and this is also due to the fact that the geographic areas interested in mycotoxin outbreaks are widening. Among the different pre- and post-harvest strategies that may be applied to prevent fungal and/or aflatoxin contamination, fungicides still play a prominent role; however, despite of countless efforts, to date the problem of food and feed contamination remains unsolved, since the essential factors that affect aflatoxins production are various and hardly to handle as a whole. In this scenario, the exploitation of bioactive natural sources to obtain new agents presenting novel mechanisms of action may represent a successful strategy to minimize, at the same time, aflatoxin contamination and the use of toxic pesticides. The Aflatox^® Project was aimed at the development of new-generation inhibitors of aflatoxigenic Aspergillus spp. proliferation and toxin production, through the modification of naturally occurring molecules: a panel of 177 compounds, belonging to the thiosemicarbazones class, have been synthesized and screened for their antifungal and anti-aflatoxigenic potential. The most effective compounds, selected as the best candidates as aflatoxin containment agents, were also evaluated in terms of cytotoxicity, genotoxicity and epi-genotoxicity to exclude potential harmful effect on the human health, the plants on which fungi grow and the whole ecosystem.     

Hydroxyls nests in defective silicalites and strained structures derived upon dehydroxylation: vibrational properties and theoretical modelling

Defective silicalite, an efficient and selective catalyst in the gas phase Beckmann rearrangement reaction, has been characterised by infrared spectroscopy and by molecular modelling techniques. We report a detailed IR study on the effect of outgassing treatments at increasing temperature on silanols bands and on framework modes. The effect of a temperature decrease up to 100 K (during the IR measurement) on the H-bonding interactions has also been investigated. The interaction of silanols with mesitylene, a probe molecule which cannot penetrate the channels, has been studied in order to distinguish between internal and external OH groups. Molecular mechanics and ab initio methods have also been used to model the structure and the vibrational features of a properly designed nest in order to support the assignments of the IR spectrum.     

Enzymatic hydrolysis of cotton wastes. Effect of γ-ray pretreatments

Textile cotton wastes were irradiated with γ-rays at different doses and solubilized with 10% NaOH solutions. The regenerated products underwent enzymatic hydrolysis and their degradation pattern was compared with that of the cotton celluloses untreated or treated only with γ-rays. The different samples were characterized by measurements of X-ray diffraction, degree of polymerization, moisture regain and water retention. The structural and morphological modifications induced by the pretreatments were correlated with the variations of kinetic parameters, particularly the V_max/K_m ratio, defined as specificity constant. Only the combination of irradiation with dissolution pretreatments gave a considerable increase of hydrolysis reaction effectiveness.     

Probabilistic consensus clustering using evidence accumulation

Machine Learning - Clustering ensemble methods produce a consensus partition of a set of data points by combining the results of a collection of base clustering algorithms. In the evidence...