Changes in the cerebellar cytoarchitecture of hibernating hedgehog Erinaceus europaeus L. (Mammalia): an immunocytochemical approach

Hibernation is an amazing animal strategy to survive when the environmental temperature is very low and food resources are scarce. Successful hibernation requires a variety of complex biological adaptations, in which the brain plays a central regulatory role. Currently, little information is available regarding the morphology and functional activity of specific neurons within the cerebellar cytoarchitecture of hibernating animals. In the present study, we investigated the immunohistochemical expression of essential proteins in the cerebellum of a mammalian hibernator (i.e. hedgehog Erinaceus europaeus L.), focusing on (i) Purkinje neurons, the sole output cells of the cerebellar cortex; (ii) selected neurotransmitters involved in hibernation processes; (iii) intracellular calcium homeostasis, considering that calcium is also an important regulator of neurotransmission mechanisms; and (iv) cytoskeletal proteins, involved in maintenance of neuronal shape and axon calibre. Specifically, we studied in situ immunocytochemical changes during the torpor state of hibernation (November–March) versus the activity phase (April–September). We employed different selected markers, i.e. glutamic acid decarboxylase (GAD67) and postsynaptic glutamate ionotropic receptor GluR2-3, different calcium-binding proteins (i.e. calbindin, parvalbumin and calretinin) and cytoskeletal components (i.e. pNF-H and MAP2). In summary, our data in hibernating animals demonstrated: (i) downregulation of GAD67, indicating loss/changes of synaptic contacts on Purkinje somata and dendrites; (ii) GluR2-3 upregulation in Purkinje neurons, with a drastic decrease of calbindin expression; and (iii) decrease of normal mechanisms regulating intracellular calcium homeostasis. We also found a decrease/modification in the distribution of cytoskeletal proteins, particularly evident for pNF-H. Changes in the functional activity of Purkinje cells were accompanied by some morphological dendrite alterations, signs of degeneration in cell somata and flattened basket pinceaux at the Purkinje axon hillock. These findings confirm that hibernation makes heterothermic animals a valuable model to study physiological adaptations to adverse conditions, and also for understanding cellular and molecular mechanisms aimed at preserving mammalian organs from full degeneration and death

Pt(IV)Ac-POA: new platinum compound Induced caspase independent apoptosis In B50 neuroblastoma stem cells

Neuroblastoma is a tumour that affects adults and children, characterized by a stem cells component. To date, cisplatin is the main antitumor agent used in the clinical treatment of this tumour; however, it induces side effects such as neurotoxicity in healthy cells and induces chemo resistance to therapy in cancer cells. New platinum-based compounds, platinum (II) have recently been synthesized, and due to their chemical characteristics, they are able to identify new cellular targets. These complexes act as prodrugs and performing their cytotoxic effect as platinum (II) after a reduction reaction within the hypoxic tumour cells. Among these prodrugs, Pt(IV)Ac-POA appears to be very promising, thanks to the presence of ligand (2 propinyl)octanoic acid (POA), which acts as an inhibitor of histone deacetylase (HDACi) and leads to the increase of histone acetylation, decreasing the interactions between histone and DNA, so as to produce chemo-sensitization to DNA-damaging agents. The greater cytotoxic effect of Pt(IV)Ac-POA on tumour cells, would therefore be mainly due to the mechanism of inhibition of histone deacetylase, which would increase the accessibility of DNA to platination mechanisms that induce cell death. In this study the results show that Pt(IV)Ac-POA, used at a concentration ten times lower than cisplatin, can induce apoptosis in B50 cells in culture both through the intrinsic pathway and through the independent caspase pathway. The data, obtained by immunohistochemical techniques in fluorescence microscopy, show that treatment with Pt(IV)Ac-POA has a greater proapoptotic effect on stem cells compared to the cisplatin standard treatment

Searching for a Longevity Food, We Bump into Hericium erinaceus Primordium Rich in Ergothioneine: The “Longevity Vitamin” Improves Locomotor Performances during Aging

Phenotypic frailty is characterized by a progressive decline in physical functioning. During ageing, morphological and functional alterations involve the brain, and chief theories involve ox-idative stress, free radical accumulation, and reduced antioxidant defenses as the most implicated mechanisms. From boosting the immune system to fighting senescence, medicinal mushrooms have been found to have a number of health and longevity benefits. Among them, Hericium erinaceus (He) has been demonstrated to display a variety of physiological effects, including anti-aging properties. Thus, He represents an attractive natural source for developing novel medicines and functional foods, based on the identification of its active ingredients and metabolites. Particularly, H. erinaceus primordium (He2) extract contains a high amount of Ergothioneine (ERGO), the longevity vitamin. Herein, we revealed the preventive effect of ERGO-rich He2 extract in a preclinical model, focusing on locomotor decline during ageing monitored through spontaneous behavioral test. This effect was accompanied by a significant decrease in some oxidative stress markers (NOS2, COX2) paralleled by an increase in P53, showed in cerebellar cortex cells and fibres by immunohistochemistry. In summary, we demonstrated the neuro-protective and preventive effects of He2 extract during aging, probably due to its peculiarly high ERGO content

MIRTO: an open-source robotic platform for education

This paper introduces the MIddlesex RoboTic platfOrm (MIRTO), an open-source platform that has been used for teaching First Year Computer Science students since the academic year 2013/2014, with the aim of providing a physical manifestation of Software Engineering concepts that are often delivered using only abstract or synthetic case studies. In this paper we provide a detailed description of the platform, whose hardware specifications and software libraries are all released open source; we describe a number of teaching usages of the platform, report students’ projects, and evaluate some of its aspects in terms of effectiveness, usability, and maintenance

Aggiornamenti e novità sulle conoscenze di Pancratium maritimum (Amaryllidaceae)

Pancratium maritimum L., bulbosa perenne degli ambienti dunali costieri, è un taxon relativamente recente se comparato con le specie del genere a gravitazione mediterranea. Gli studi noti su questa specie hanno analizzato vari aspetti della biologia (riproduttivi, biochimici, filogenetici, genetici, ecc.). Tuttavia, esistono ancora lacune scientifiche, tra cui l’assenza di studi su ampio areale di genetica di conservazione e sugli adattamenti eco-morfofisiologici alle condizioni di stress. Per ampliare le conoscenze su P. maritimum, la Fondazione Nando Peretti nel novembre 2012 ha finanziato un progetto triennale sulla conoscenza e la salvaguardia di questa specie (Progetto 2012-83). In questo contributo si presentano i risultati ottenuti per le diverse linee di ricerca finora affrontate, che vanno dalle analisi ad ampio areale dei popolamenti, usando un approccio sia genetico che GIS, alla caratterizzazione morfologica e eco-fisiologica di varie popolazioni

WNT inhibitory activity of malus pumila miller cv annurca and malus domestica cv limoncella apple extracts on human colon-rectal cells carrying familial adenomatous polyposis mutations

Inhibitors of the Wingless-related Integration site (WNT)/β-catenin pathway have recently been under consideration as potential chemopreventive agents against Familial Adenomatous Polyposis (FAP). This autosomal-dominant syndrome is caused by germline mutations in the gene coding for the protein APC and leads to hyperactivation of the WNT/β-catenin signaling pathway, uncontrolled intestinal cell proliferation and formation of adenocarcinomas. The aim of the present work was to: (i) test, on in vitro cultures of cells carrying FAP mutations and on ex vivo biopsies of FAP patients, the WNT inhibitory activity of extracts from two common southern Italian apples, Malus pumila Miller cv. ‘Annurca’ and Malus domestica cv ‘Limoncella’; (ii) identify the mechanisms underpinning their activities and; (iii) evaluate their potency upon gastrointestinal digestion. We here show that both Annurca and Limoncella apple extracts act as WNT inhibitors, mostly thanks to their polyphenolic contents. They inhibit the pathway in colon cells carrying FAP mutations with active dilutions falling in ranges close to consumer-relevant concentrations. Food-grade manufacturing of apple extracts increases their WNT inhibitory activity as result of the conversion of quercetin glycosides into the aglycone quercetin, a potent WNT inhibitor absent in the fresh fruit extract. However, in vitro simulated gastrointestinal digestion severely affected WNT inhibitory activity of apple extracts, as result of a loss of polyphenols. In conclusion, our results show that apple extracts inhibit the WNT pathway in colon cells carrying FAP mutations and represent a potential nutraceutical alternative for the treatment of this pathology. Enteric coating is advisable to preserve the activity of the extracts in the colon-rectal section of the digestive tract. © 2017 by the authors. Licensee MDPI, Basel, Switzerland

Identification and functional characterization of polymorphisms in human cyclooxygenase-1 (PTGS1)

Cyclooxygenase-1 (COX-1, PTGS1) catalyzes the conversion of arachidonic acid to prostaglandin H2, which is subsequently metabolized to various biologically active prostaglandins. We sought to identify and characterize the functional relevance of genetic polymorphisms in PTGS1

Epidemiology of reported Yersinia enterocolitica infections in Germany, 2001-2008

<p>Abstract</p> <p>Background</p> <p>Yersiniosis is the third most common zoonotic bacterial disease in Germany and the European Union. Sequelae of <it>Yersinia enterocolitica </it>infections, such as reactive arthritis, have been reported. Consumption of pork and its products, especially eaten raw or undercooked, is an important risk factor of yersiniosis. Infection with <it>Y. enterocolitica </it>is notifiable through the national surveillance system for infectious diseases in Germany and several thousands of cases are being reported each year. We present recent data on the epidemiology of reported yersiniosis in Germany.</p> <p>Methods</p> <p>Surveillance data on yersiniosis, accessed through the national level database (SurvNet), were analyzed with regard to time trends, demographical and geographical distribution, serotypes, and hospitalization, for the time period 2001-2008.</p> <p>Results</p> <p>A total of 47,627 cases of yersiniosis were reported. The mean annual incidence of yersiniosis was 7.2/100,000 population. A downward trend in the number of reportable cases has occurred since 2002. Almost all <it>Y. enterocolitica </it>infections were reported as single cases, i.e., with no apparent links to other cases. The number of reported infections showed substantially less seasonal variation than in other zoonotic enteric diseases. The incidence was highest in children under five years (58/100,000 population), in particular in one-year-old children (108/100,000 population). Almost 97% of infections were acquired domestically. High incidences occurred in the eastern German federal states Thuringia, Saxony, and Saxony-Anhalt. Differences in incidences across federal states were driven primarily by incidence differences in children under five years. Hospitalization was reported for 17% of cases, the proportion being highest among teenagers. Almost 90% of <it>Y. enterocolitica </it>strains were diagnosed as serotype O:3, which is the serotype most frequently isolated from pigs.</p> <p>Conclusions</p> <p>Yersiniosis is a zoonotic foodborne disease of relevance to public health in Germany because of its high incidence and risk for sequelae. The incidence of reported yersiniosis in Germany varies markedly from state to state, mainly due to incidence difference among young children. More research efforts should be directed towards the elucidation of risk factors of yersiniosis in this age group.</p

Timing the multiple cell death pathways initiated by Rose Bengal acetate photodynamic therapy

Rose Bengal acetate photodynamic therapy (RBAc–PDT) induced multiple cell death pathways in HeLa cells through ROS and ER stress. Indeed, apoptosis was the first preferred mechanism of death, and it was triggered by at least four different pathways, whose independent temporal activation ensures cell killing when one or several of the pathways are inactivated. Apoptosis occurred as early as 1 h after PDT through activation of intrinsic pathways, followed by activation of extrinsic, caspase-12-dependent and caspase-independent pathways, and by autophagy. The onset of the different apoptotic pathways and autophagy, that in our system had a pro-death role, was timed by determining the levels of caspases 9, 8, 3 and 12; Bcl-2 family; Hsp70; LC3B; GRP78 and phospho-eIF2α proteins. Interestingly, inhibition of one pathway, that is, caspase-9 (Z-LEHD-FMK), caspase-8 (Z-IETD-FMK), pan-caspases (Z-VAD-FMK), autophagy (3-MA) and necrosis (Nec-1), did not impair the activation of the others, suggesting that the independent onset of the different apoptotic pathways and autophagy did not occur in a subordinated manner. Altogether, our data indicate RBAc as a powerful photosensitiser that induces a prolonged cytotoxicity and time-related cell death onset by signals originating from or converging on almost all intracellular organelles. The fact that cancer cells can die through different mechanisms is a relevant clue in the choice and design of anticancer PDT